Postconditioning with a CpG containing oligodeoxynucleotide ameliorates myocardial infarction in a murine closed-chest model.
Toll-like receptor (TLR)9 ligand CpG-oligodeoxynucleotide (CpG-ODN) exerts preconditioning in myocardial ischemia/reperfusion. We hypothesized a postconditioning effect of CpG-ODN in a murine closed-chest model of myocardial infarction. C57BL/6 (12 weeks, male, WT) mice were instrumented at the left anterior descending artery, then allowed 5d of recovery before 30 min ischemia. Treatments comprised: 1) PBS: 250 μl phosphate buffer solution intraperitoneally 5 min before reperfusion and 2) IPC (ischemic postconditioning): 3 twenty-second reperfusion and occlusion episodes at the end of ischemia 3) CpG-ODN: 1668 thioate 0.2 μmol/kg BW intraperitoneally 5 min before reperfusion. Infarct size was assessed via triphenyltetrazolium chloride (TTC) staining after 2 and 24h reperfusion. Myocardial mRNA-expression of cytokines was measured using real-time PCR after 2h reperfusion. Phosphatidylinositol-3 kinase (PI3K)-inhibitor wortmannin was injected intraperitoneally in WT 15 min before postconditioning and PBS in each group. Cardiac function in WT was assessed with a left-ventricular pressure-volume catheter at 24h reperfusion. Following 30 min ischemia and 2h reperfusion, infarct size was diminished by 90% in WT postconditioned with CpG-ODN (2.4 ± 1.55 IS/AAR%) and IPC (1.98 ± 1.03 IS/AAR%) compared to PBS mice (23.2 ± 3.97 IS/AAR%). Infarct size increased following 24h reperfusion but the differences remained robust. Expression of TNF-α and IL-10 was increased in CpG-ODN. Wortmannin abolished the postconditioning effect of CpG-ODN and IPC. Ejection fraction and preload-recruitable stroke work were significantly greater in CpG-ODN mice. CpG-ODN confers postconditioning via activation of TLR9. Cardiac function is preserved following CpG-ODN postconditioning. The PI3K -inhibitor wortmannin attenuates CpG-ODN postconditioning.


Myricitrin blocks activation of NF-κB and MAPK signaling pathways to protect nigrostriatum neuron in LPS-stimulated mice.
Myricitrin, a bioactive and natural flavonoids, is well known for its anti-inflammatory and antioxidant properties. However, the anti-neuroinflammation and possible mechanism has not been fully elucidated. Therefore, the present study was to investigate the possible mechanism of its neuroprotection and anti-neuroinflammation in the nigrostriatum of LPS-stimulated mice. The results showed that myricitrin improved neuron injury and raised the expressions of PSD-95 protein and TH protein in the nigrostriatum of LPS-stimulated mice. In addition, myricitrin decreased the production of pro-inflammatory factors including IL-1β, IL-6 and TNFα, decreased the level of chemokine MCP-1, and suppressed the expressions of COX-2 and iNOS. Meanwhile, myricitrin suppressed HMGB1, TLR4, and MyD88 expression in the nigrostriatum of LPS-stimulated mice. Furthermore, myricitrin inhibited NF-κB and MAPK signaling pathways activated by LPS. In conclusion, our studies suggest that myricitrin blocks activation of protects NF-κB and MAPK signaling pathways to nigrostiatum neuron from injury in LPS-stimulated mice and is beneficial to treatment nigrostriatum inflammation of PD.


Oligomerization-driven MLKL ubiquitylation antagonizes necroptosis.
Mixed lineage kinase domain-like (MLKL) is the executioner in the caspase-independent form of programmed cell death called necroptosis. Receptor-interacting serine/threonine protein kinase 3 (RIPK3) phosphorylates MLKL, triggering MLKL oligomerization, membrane translocation and membrane disruption. MLKL also undergoes ubiquitylation during necroptosis, yet neither the mechanism nor the significance of this event has been demonstrated. Here, we show that necroptosis-specific multi-mono-ubiquitylation of MLKL occurs following its activation and oligomerization. Ubiquitylated MLKL accumulates in a digitonin-insoluble cell fraction comprising organellar and plasma membranes and protein aggregates. Appearance of this ubiquitylated MLKL form can be reduced by expression of a plasma membrane-located deubiquitylating enzyme. Oligomerization-induced MLKL ubiquitylation occurs on at least four separate lysine residues and correlates with its proteasome- and lysosome-dependent turnover. Using a MLKL-DUB fusion strategy, we show that constitutive removal of ubiquitin from MLKL licences MLKL auto-activation independent of necroptosis signalling in mouse and human cells. Therefore, in addition to the role of ubiquitylation in the kinetic regulation of MLKL-induced death following an exogenous necroptotic stimulus, it also contributes to restraining basal levels of activated MLKL to avoid unwanted cell death.


Endoscopic ultrasound with double-balloon endoscopy for the diagnosis of inverted Meckel's diverticulum: a case report.
Inverted Meckel's diverticulum has usually been misdiagnosed in the cases based on computed tomography images presented in the literature. The final diagnosis was made intra-operatively or by pathology reports after surgery. Despite this, preoperative diagnosis could be made successfully by using endoscopic ultrasound with double-balloon endoscopy prior to surgery. A 60-year-old Japanese woman with severe anemia complained of several episodes of black stool over the preceding 2 years. Abdominal computed tomography showed a 3.0-cm low-density tumor in the ileum, suggesting a diagnosis of intestinal lipoma. Examination of the tumor by endoscopic ultrasound with double-balloon endoscopy revealed a hypo-echoic layer corresponding to the muscularis propria, and a hyper-echoic layer corresponding to the fat tissue. These findings, which suggested that the tumor included areas outside the intestinal serosa, are not typical for a lipoma, despite the existence of a hyper-echoic layer corresponding to fatty tissue. We then considered a diagnosis of inverted Meckel's diverticulum. Lipoma and inverted Meckel's diverticulum are difficult to differentially diagnose by computed tomography. Polypectomy is the preferred therapeutic approach when a lipoma is present; however, polypectomy in a patient with Meckel's diverticulum requires full-thickness resection. Situations where polypectomy is performed without preparing for full-thickness resection can be avoided by first making a precise diagnosis using double-balloon endoscopy and endoscopic ultrasound.


Addressing social determinants of health in community pharmacy: Innovative opportunities and practice models.
Social determinants of health (SDoH) account for up to 90% of health outcomes, whereas medical care accounts for only 10%-15%; despite this disparity, only 24% of hospitals and 16% of physician practices screen for the 5 social needs. Community-embedded and highly accessible, pharmacies are uniquely positioned to connect individuals to local community and social resources and thereby address SDoH. In this article, we explore novel community pharmacy practice models that address SDoH, provide real-world examples of these models, and discuss pathways for reimbursement and sustainability. A number of innovative community pharmacy practice models that focus on social issues are currently being explored. These include integrating community health workers (CHWs) or SDoH specialists, wherein CHWs are frontline public health workers who can effectively bridge the health care system and their community, whereas SDoH specialists are pharmacy team members trained with substantial SDoH knowledge and how to use it to connect pharmacy patients to community resources. Three community pharmacy networks have implemented pilot programs using either a CHW or SDoH specialist model. An essential component for program success in all cases has been partnership development and increased interdependence between the pharmacies, local community organizations, and the public health sector. New payment models and financial incentives will be necessary to expand and sustain these programs. A potential Approach may be the use of Z codes, a subset of ICD-10-CM codes specific to assessing SDoH. Although opportunities are developing for community pharmacies to play a major role in sustainably addressing SDoH, additional work is needed before there is a widespread acceptance of pharmacies becoming service referral destinations for patients with social needs. Evaluation of these models on a wider scale will be necessary to fully evaluate their effectiveness, costs, and implementation within different community pharmacy settings.


Kidney transplantation in patients with end stage congenital renal disease. Report of eighteen cases and review of the Organ Transplant Registry.
Eighteen patients with end stage congenital renal disease requiring kidney transplantation constituted 12 per cent of the transplantation recipients at this institution over the past six years. The post-transplantation course in this group was remarkably satisfactory, with a 94 per cent graft survival at three years. In addition, we analyzed survival data from over 9,900 patients in the Organ Transplant Registry and demonstrated that transplant recipients with end stage congenital renal disease have equal or better five year patient survival compared with those with acquired end stage renal disease. Only those patients with adult polycystic disease had a less satisfactory prognosis, probably because of age-related factors. In contrast, there were few statistical correlations between renal allograft survival, age, and original disease.


Identification of educational objectives for obstetrics and gynecology residents in the ambulatory setting.
The primary objective of this study was to identify specific educational goals for obstetrics and gynecology residents in the ambulatory setting. A cross-sectional study of current practice patterns in primary care, benign gynecology, and office procedures was performed with mailings to local private practice obstetrician-gynecologists. Questions for the anonymous written survey were generated using the Council on Resident Education in Obstetrics and Gynecology educational objectives. Telephone interviews with staff from billing offices confirmed practitioner responses. Of 88 practitioners, 43 (49%) responded. Diagnoses made in the office correlated well with the topics considered important for resident knowledge. Most important primary care diagnoses were depression and abdominal pain; important gynecologic diagnoses were abnormal uterine bleeding, chronic pelvic pain, contraception, and vulvovaginal infection. This study offers a valid, practical foundation for developing a focused ambulatory resident education program based on current outpatient obstetrics-gynecology practice patterns.


[The feeding characteristics of mosquito larvae of the genus Anopheles].
Studies carried out on 6 species of malaria mosquitos revealed a relationship between the larval activity to scratch the substrate and their morphological and physiological features and also their development at different depth of feed immersion. The development of scratching capacity was compared in different species of Anopheles and Cellia subgenera.


Development ofDentalium following removal of D-quadrant blastomeres at successive cleavage stages.
Each primary micromere and macromere of the D-quadrant ofDentalium was deleted, through the mesentoblast stage, to investigate the way in which the polar lobe cytoplasm exerts its influence on development.-D and -1D embryos form an apical tuft but no posttrochal structures.-2D embryos form an apical tuft and a reduced posttrochal region without a shell. -3D and -4D are externally similar to control embryos. -1d embryos and -1c embryos have an apical tuft with a reduced number of cilia. Embryos in which both 1c and 1d are deleted lack the apical tuft.-2d embryos lack shell and most other posstrochal structures. -3d and-4d embryos appear externally equivalent to controls.The polar lobe cytoplasm exerts its influence sequentially, and as inIlyanassa the maximal effect is at the third quartet stage.


Eye lens ageing in the dogfish (Mustelus canis).
1. Age-related alterations in the distribution of water-soluble, high molecular weight (colloidal), and water-insoluble proteins of the lens of smooth dogfish (Mustelus canis) were measured. 2. The ages of these animals ranged approx from 2 to 50 yr, during which time the lenses grew from 100 to 1500 mg (wet wt). The lenses contained approx 50% water. 3. Water-insoluble protein accumulated to a level greater than 50% of the total proteins by the time the animals reached maturity. The lenses of other animals, such as mammals and humans, would be opaque if they had a similar insoluble protein content. 4. Each protein fraction contained the same protein chains (mol. et 1900-25,000 daltons), as observed by SDS polyacrylamide gel electrophoresis, except the water-insoluble fraction, which seemed to contain several extra protein chains with higher molecular weights, which represent fiber cell membrane components. 5. Further purification of these fiber cell membranes indicated that their protein chain makeup was mainly from the same low molecular weight chains present in the soluble and high molecular weight colloidal proteins.


Autoperfusion balloon catheter for treatment of vertebral artery stenosis.
Percutaneous transluminal angioplasty (PTA) of the vertebral artery was performed with an autoperfusion balloon catheter in five patients. There were no complications in the form of embolic episodes or neurological deficits due to brain ischaemia during inflation. In critical cases with insufficient collateral circulation during temporary occlusion, the use of an autoperfusion balloon catheter may expand the indications for PTA in patients with ischaemic cerebrovascular disease.


Bioinformatics analysis of gene expression profiles in B cells of postmenopausal osteoporosis patients.
The aim of this study was to gain a better understanding of the molecular mechanisms and identify more critical genes associated with the pathogenesis of postmenopausal osteoporosis (PMOP). Microarray data of GSE13850 were download from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) were identified either in B cells from postmenopausal female nonsmokers with high bone mineral density (BMD) compared with those with low BMD (defined as DEG1 group) or in B cells from postmenopausal female smokers with high BMD compared with postmenopausal female nonsmokers with high BMD (defined as DEG2 group). Gene ontology and immune-related functional enrichment analysis of DEGs were performed. Additionally, the protein-protein interaction network of all DEGs was constructed and subnetworks of the hub genes were extracted. A total of 51 DEGs were identified in the DEG1 group, including 30 up- and 21 downregulated genes. Besides, 86 DEGs were identified in the DEG2 group, of which 46 were upregulated and 40 were downregulated. Immune enrichment analysis showed DEGs were mainly enriched in functions of CD molecules and chemokines and receptor, and the upregulated gene interleukin 4 receptor (IL-4R) was significantly enriched. Moreover, guanine nucleotide-binding protein G (GNAI2), filamin A alpha (FLNA), and transforming growth factor-β1 (TGFB1) were hub proteins in the protein-protein interaction network. IL-4R, GNAI2, FLNA, and TGFB1 may be potential target genes associated with the pathogenesis of PMOP. In particular, FLNA, and TGFB1 may be affected by smoking, a risk factor of PMOP.


Unique system of care issues and challenges in serving children under age 3 and their families.
This article addresses the primary modifications necessary for system change to better meet the mental health needs of children under the age of three. The role of risk and resiliency factors in the young child, family and community and the necessity for a comprehensive community infant-family mental health system with a focus on the whole family are addressed. Barriers to care within early childhood systems of care are examined, including stigma, community referral and collaboration, diagnostic concerns during infancy, issues around family engagement, empowerment and partnership, funding of comprehensive and well coordinated infant-family services, workforce capacity and evaluation. Recommendations for implementation of system changes at the community and federal levels are proposed.


Metabolic oxidative stress induced by a combination of 2-DG and 6-AN enhances radiation damage selectively in malignant cells via non-coordinated expression of antioxidant enzymes.
Our earlier studies have shown that simultaneous inhibition of glycolysis and pentose phosphate pathway using 2-deoxy-d-glucose (2-DG, an inhibitor of glycolysis) and 6-aminonicotinamide (6-AN, an inhibitor of pentose phosphate pathway) lead to metabolic oxidative stress (MOS), resulting in radiosensitization in malignant cells. Present study was carried out to investigate the effects of 2-DG and 6-AN on intricately regulated endogenous antioxidant defense against MOS during radiosensitization by this combination. Two human tumor cell lines {Head and Neck Squamous carcinoma (KB) and Glioma (BMG-1)} and one non-malignantly transformed cell line (human embryonic kidney, HEK) were used in this study. The presence of 2-DG and 6-AN (added just before irradiation) for 4h, significantly decreased the clonogenicity and metabolic viability of KB and BMG-1 cell lines, while no significant change was seen in HEK cells. Accumulation of ROS was observed only in malignant cell lines, which displayed a compromised redox status evident from enhanced NADP(+)/NADPH and GSSG/GSH ratios and a concomitant decrease in glutathione reductase level and activity at 24h following treatment. The levels and activities of Cu, Zn-SOD and Mn-SOD increased with MOS and were accompanied by a decreased GPx and unaltered catalase activity and level. These results suggest that non-coordinated expression of antioxidant defense, besides compromised redox status, led to selective radiosensitization in the malignant cells.


Effectiveness and safety of wheelchair skills training program in improving the wheelchair skills capacity: a systematic review.
To comprehensively assess the effectiveness and safety of wheelchair skills training program in improving wheelchair skills capacity. PubMed, OVID, EBSCO, ScienceDirect, Web of Science, CINAHL, Cochrane Library, Google Scholar, and China Knowledge Resource Integrated Database were searched up to March 2017. Controlled clinical trials that compared a wheelchair skills training program with a control group that received other interventions and used the wheelchair skills test scores to evaluate wheelchair skills capacity were included. Two authors independently screened articles, extracted data, and assessed the methodological quality using the Cochrane risk-of-bias tool in randomized controlled trial (RCT) and methodological index for non-randomized studies. The data results of wheelchair skills test scores were extracted. Data from 455 individuals in 10 RCTs and from 140 participants in seven non-randomized studies were included for meta-analysis using Stata version 12.0 (Stata Corporation, College Station, TX, USA). In the short term (immediately to one week) post-intervention, relative to a control group, manual wheelchair skills training could increase the total wheelchair skills test scores by 13.26% in RCTs (95% confidence interval (CI), 6.19%-20.34%; P < 0.001) and by 23.44% in non-randomized studies (95% CI, 13.98%-32.90%; P < 0.001). Few adverse events occurred during training; however, compared with a control group, evidence was insufficient to support the effectiveness of powered wheelchair skills training and the long-term (3-12 months) advantage of manual wheelchair skills training ( P = 0.755). The limited evidence suggests that wheelchair skills training program is beneficial in the short term, but its long-term effects remain unclear.


Eye movement influences on coupled and decoupled eye-hand coordination tasks.
Visually guided reaching precision and accuracy depend on the level of coupling between movements of the eyes and hand. In the present study, participants performed central fixations and either saccadic or smooth pursuit eye movements during fast and accurate reaching tasks involving eye-hand coupling and decoupling to better understand type of eye movement influence over upper limb control. Some eye-hand coupling and decoupling tasks also included hand reversals, where the hand moves away from the target to direct a cursor toward the target to account for various levels of hand-cursor and eye-cursor coupling. Regardless of eye-movement type, eye-hand-cursor coupling produced an endpoint accuracy advantage over decoupling. Use of hand reversal decreased peak speed and increased response time of the hand, whether considering fixation or a given eye movement. Use of smooth pursuit slowed hand movements relative to saccades, yet improved endpoint accuracy. Compared to central fixations, using smooth pursuit also slowed hand movements, while using saccades decreased, thus improved, hand reaction times. Data suggest an advantage, when using smooth pursuit to track the hand movement for the greatest endpoint accuracy, an advantage when using saccades for the fastest movements, and an eye-hand coupling advantage when using saccades for the shortest reactions. Researchers should provide clear eye-movement instructions for participants and/or monitor the eyes when assessing similar upper limb control to account for possible differences in eye movements used. Moreover, the type of eye movement chosen for participants should correspond to the primary goal of the task.


[Effect of radix Salviae miltiorrhizae on production of free radical products from lung during cardiopulmonary bypass operation].
Thirty patients of heart valve replacement were randomly divided into two groups, one as Radix Salviae Miltiorrhizae (RSM) treatment group and the other as control group. Blood samples were taken from right atrium (RA) and left atrium (LA) before cardiopulmonary bypass operation (CPB) and 10 min, 45 min, 2 hours, 6 hours after the removal of the aortic cross-clamp to observe the level of peroxidation product and the changes in leucocyte count. The results showed that the number of leucocyte in LA was smaller than that in RA and the amount of peroxidation product in LA was higher than that in RA (P < 0.01-0.05) during the early phase of the lung reperfusion in control group. There was no significant difference (P > 0.05) in the amount of peroxidation product and the number of leucocyte between LA and RA in RSM treatment group. The results demonstrated that RSM could markedly prevent the lung leucocyte aggregation and reduce the production of lung free radical products. It suggested that the effect of RSM of reducing the production of lung free radical products was related to its effect of preventing the lung leucocyte aggregation.


[Comparative action of chronic injection of alpha toxin from Naja nigricolis and of botulinum toxin A on the development of chick embryo dorsal and ventral spinal roots].
The chronic injection of high doses of alpha toxin from Naja nigricollis or botulinum toxin type A in the yolk sack of developing Chick embryos causes a selective decrease of the number of fibers in the ventral roots of the spinal nerves; at the concentrations needed this reduction is more important with the alpha toxin than with the botulinum toxin.


Characterization of Flexible Amorphous Silicon Thin-Film Transistor-Based Detectors with Positive-Intrinsic-Negative Diode in Radiography.
Low-dose exposure and work convenience are required for mobile X-ray systems during the COVID-19 pandemic. We investigated a novel X-ray detector (FXRD-4343FAW, VIEWORKS, Anyang, Korea) composed of a thin-film transistor based on amorphous silicon with a flexible plastic substrate. This detector is composed of a thallium-doped cesium iodide scintillator with a pixel size of 99 μm, pixel matrix of 4316 × 4316, and weight of 2.95 kg. The proposed detector has the advantages of high-noise characteristics and low weight, which provide patients and workers with an advantage in terms of the dose and work efficiency, respectively. We performed a quantitative evaluation and an experiment to demonstrate its viability. The modulation transfer function, noise power spectrum, and detective quantum efficiency were identified using the proposed and comparative detectors, according to the International Electrotechnical Commission protocol. Additionally, the contrast-to-noise ratio and coefficient of variation were investigated using a human-like phantom. Our results indicate that the proposed detector efficiently increases the image performance in terms of noise characteristics. The detailed performance evaluation demonstrated that the outcomes of the use of the proposed detector confirmed the viability of mobile X-ray devices that require low doses. Consequently, the novel FXRD-4343FAW X-ray detector is expected to improve the image quality and work convenience in extended radiography.


The mouth matters most: A functional magnetic resonance imaging study of how dogs perceive inanimate objects.
The perception and representation of objects in the world are foundational to all animals. The relative importance of objects' physical properties versus how the objects are interacted with continues to be debated. Neural evidence in humans and nonhuman primates suggests animate-inanimate and face-body dimensions of objects are represented in the temporal cortex. However, because primates have opposable thumbs and interact with objects in similar ways, the question remains as to whether this similarity represents the evolution of a common cognitive process or whether it reflects a similarity of physical interaction. Here, we used functional magnetic resonance imaging (fMRI) in dogs to test whether the type of interaction affects object processing in an animal that interacts primarily with its mouth. In Study 1, we identified object-processing regions of cortex by having dogs passively view movies of faces and objects. In Study 2, dogs were trained to interact with two new objects with either the mouth or the paw. Then, we measured responsivity in the object regions to the presentation of these objects. Mouth-objects elicited significantly greater activity in object regions than paw-objects. Mouth-objects were also associated with activity in somatosensory cortex, suggesting dogs were anticipating mouthing interactions. These findings suggest that object perception in dogs is affected by how dogs expect to interact with familiar objects.


Awareness of radiation hazards in patients attending radiology departments.
Evaluating the knowledge of patients attending radiology departments regarding ionizing radiation used in medical imaging and its associated hazards can provide knowledge of the patient's awareness level of the associated risk of the radiation used in medical imaging. The aims of this study were to evaluate the awareness of patients regarding medical radiation types used in medical diagnostic imaging and its influence on their decision to proceed with that procedure. Over an 8-months period, a total of 418 patients, 48% Men and 52% Women, presenting for diagnostic imaging in the department of radiology, completed a 15-point questionnaire. The questionnaire included demographic and radiation awareness sections. Less than 32% of the participants had a potential risk of radiation explained by the doctor before the procedure. 59% of the participants expressed that the potential risk of radiation makes them anxious; less than about 25% of the participants showed that the potential risk of radiation affects their decision to have the procedure. Overall, the data collected from this survey indicate that there is a lack of information about radiation risk provided to the patients prior to the diagnostic procedure. Efforts should be made to ensure that patients receiving multiple medical imaging tests are aware of the radiation they are receiving.


Pyruvate-fortified fluid resuscitation improves hemodynamic stability while suppressing systemic inflammation and myocardial oxidative stress after hemorrhagic shock.
To determine whether controlled resuscitation with pyruvate-fortified Ringer's (PR) solution vs. conventional lactate Ringer's (LR) more effectively stabilizes mean arterial pressure (MAP) and suppresses myocardial inflammation postresuscitation. Goats were hemorrhaged (255 +/- 22 ml) to lower MAP to 48 +/- 1 mmHg. Next, the right femoral vessels were occluded for 90 min to model tourniquet application. Beginning at 30 min occlusion, LR or PR was infused i.v. at 10 ml/min for 90 min. The femoral occlusions were released at 60 min infusion. At 4 h postocclusion, MAP (mmHg) was increased in PR (59 +/- 4) vs. LR (47 +/- 3) resuscitated goats (p < 0.05). PR also more effectively augmented circulating HCO3 and total base excess. Nitrosative stress, detected in myocardium 4 h after LR resuscitation, was suppressed by PR. Finally, PR prevented the increase in circulating neutrophils that accompanied LR resuscitation. Relative to LR, resuscitation with PR more effectively stabilized MAP, suppressed myocardial nitrosative stress and minimized systemic inflammation after hemorrhagic shock with hindlimb ischemia-reperfusion.


The antidiarrheal and spasmolytic activities of Phyllanthus emblica are mediated through dual blockade of muscarinic receptors and Ca2+ channels.
This study was aimed at providing the possible mechanisms for the medicinal use of Phyllanthus emblica in diarrhea. The in vivo studies were conducted in mice, while isolated rabbit jejunum and guinea-pig ileum were used for the in vitro experiments. The crude extract of Phyllanthus emblica (Pe.Cr), which tested positive for alkaloids, tannins, terpenes, flavonoids, sterols and coumarins, caused inhibition of castor oil-induced diarrhea and intestinal fluid accumulation in mice at 500-700 mg/kg. In isolated rabbit jejunum, Pe.Cr relaxed carbachol (CCh) and K(+) (80 mM)-induced contractions, in a pattern similar to that of dicyclomine. The preincubation of guinea pig-ileum with Pe.Cr (0.3 mg/mL), caused a rightward parallel shift in the concentration-response curves (CRCs) of acetylcholine without suppression of the maximum response. While at the next higher concentration (1 mg/mL), it produced a non-parallel rightward shift with suppression of the maximum response, similar to that of dicyclomine, suggesting anticholinergic and Ca(2+) channel blocking (CCB)-like antispasmodic effect. The CCB-like activity was further confirmed when pretreatment of the tissue with Pe.Cr, shifted the CRCs of Ca(2+) to the right with suppression of the maximum response, similar to nifedipine or dicyclomine. The activity-directed fractions of Pe.Cr showed a combination of Ca(2+) antagonist and anticholinergic like components in all fractions but with varying potency. These results indicate that the Phyllanthus emblica fruit extract possesses antidiarrheal and spasmolytic activities, mediated possibly through dual blockade of muscarinic receptors and Ca(2+) channels, thus explaining its medicinal use in diarrhea.


Improved thin-layer chromatographic separation of 32P-postlabeled DNA adducts.
DNA adducts represent the putative initiating event in the chemical carcinogenesis process. 32P-Postlabeling is one of several assays which have been developed for the sensitive detection of DNA adducts. An integral part of the 32P-postlabeling assay is the separation of adducted nucleotides by multidirectional, multisolvent, anion-exchange polyethyleneimine-cellulose thin-layer chromatography. Standard since the introduction of this assay has been the use of high-salt, high-urea solvents for the resolution of adducts during the D3 and D4 phases of the chromatography. Urea solvents are able to separate adducts resulting from a number of chemicals, however, they are time-consuming, retain a lot of background noise, may push adducts into inadequately resolved diagonal radioactive zones, and may not separate adducts of similar structure. In this study we introduce the use of a dilute ammonium hydroxide solvent for D4 chromatography and compare it to other standard solvents such as lithium chloride-Tris.HCl-urea, sodium phosphate-Tris.HCl-urea, and isopropanol-4 M ammonium hydroxide for adduct separation, resolution, recovery, retention of background noise, and chromatography development time. We found that 0.2 M ammonium hydroxide worked well for the recovery, separation, and resolution of a wide array of adducts derived from highly lipophilic polycyclic aromatic hydrocarbons and aromatic amines. In addition, this solvent required much less time (< 1/4) as compared to the other solvents and more importantly allowed the separation of adducts which otherwise comigrated and were not visible when using the other three D4 solvents.(ABSTRACT TRUNCATED AT 250 WORDS)


Copper(I)-catalyzed asymmetric alkylation of α-imino-esters.
Asymmetric alkylation of enolates is one of the most direct and important reactions to prepare α-chiral carbonyl compounds. Except for the classical methods that rely on the use of chiral auxiliaries, asymmetric catalysis emerged as a powerful tool, especially asymmetric phase-transfer catalysis. However, in the field of transition metal catalysis, only limited success with asymmetric alkylation of enolates was achieved. Hereby, we disclose a copper(I)-catalyzed asymmetric alkylation of α-imino-esters with various alkyl halides, including allyl bromides, propargyl bromide, benzyl bromides, α-bromo carbonyl compounds, and alkyl iodides. Both linear and cyclic α-imino-esters serve as competent pronucleophiles in the alkylation, which affords α-amino acid derivatives bearing either a trisubstituted or a tetrasubstituted stereogenic carbon center in high to excellent enantioselectivity. Control experiments indicate that the α-imino-ester is activated by a chiral copper(I)-phosphine complex through coordination, thus enabling facile deprotonation to provide a stabilized copper(I)-enolate in the presence of a mild base. Finally, the mildly basic nature allows the asymmetric alkylation of chiral dipeptides with excellent both chemo- and enantioselectivities.


Synthesis and antimycobacterial activities of non-purine analogs of 6-aryl-9-benzylpurines: Imidazopyridines, pyrrolopyridines, benzimidazoles, and indoles.
6,9-Disubstituted purines and 7-deazapurines are known to be powerful inhibitors of Mycobacterium tuberculosis (Mtb) in vitro. Analogs modified in the six-membered ring (imidazopyridines, pyrrolopyridines, benzimidazoles, and indoles) were synthesized and evaluated as Mtb inhibitors. The targets were prepared by functionalization on the bicyclic heterocycle or from simple pyridines. The results reported herein, indicate that the purine N-1, but not N-3, is important for binding to the unknown target. The 3-deazapurines appears to be slightly more active compared to the parent purines and slightly less active than their 7-deazapurine isomers. Removal of both the purine N-3 and N-7 did not result in further enhanced antimycobacterial activity but the toxicity towards mammalian cells was increased. Both 3-deaza and 3,7-dideazapurines exhibited a modest activity against of the Mtb isolate in the state of non-replicating persistence.


Improved clinical effectiveness of adalimumab when initiated with clindamycin and rifampicin in hidradenitis suppurativa.
Adalimumab monotherapy for hidradenitis suppurativa (HS) is often insufficient with a maximum clinical efficacy of 60% in Hidradenitis Suppurativa Clinical Response (HiSCR) and limited effect on draining tunnels. Data suggest that adalimumab therapy could be improved by concomitant antibiotics. To compare the clinical effectiveness of adalimumab with clindamycin and rifampicin versus adalimumab monotherapy after 12 weeks. This retrospective study included patients who started adalimumab with additional clindamycin and rifampicin and patients treated with adalimumab monotherapy, matched on sex and refined Hurley score. The primary outcome measure was the difference in change in the International Hidradenitis Suppurativa Severity Score System (IHS4) at 12 weeks. In total, 62 patients were included in the combination therapy group (n = 31) and adalimumab monotherapy group (n = 31), showing comparable IHS4 scores; 32.5 versus 29, p = 0.87 at baseline respectively. The combination therapy demonstrated greater clinical effectiveness expressed in median IHS4 improvement (-20 vs. -9, p < 0.001), IHS4-55 (74% vs. 36%, p = 0.002), median draining tunnel reduction (-4 vs. -2, p < 0.001) and pain response (47% vs. 27%, p = 0.02). Adalimumab initiated with clindamycin and rifampicin shows greater clinical effectiveness than adalimumab monotherapy. An important difference in effect was observed in the decrease of draining tunnels, addressing a serious limitation of adalimumab monotherapy.


Autism spectrum disorder recognition based on multi-view ensemble learning with multi-site fMRI.
Autism spectrum disorders (ASD) is a neurodevelopmental disorder that causes repetitive stereotyped behavior and social difficulties, early diagnosis and intervention are beneficial to improve treatment effect. Although multi-site data expand sample size, they suffer from inter-site heterogeneitys, which degrades the performance of identitying ASD from normal controls (NC). To solve the problem, in this paper a multi-view ensemble learning network based on deep learning is proposed to improve the classification performance with multi-site functional MRI (fMRI). Specifically, the LSTM-Conv model was firstly proposed to obtain dynamic spatiotemporal features of the mean time series of fMRI data; then the low/high-level brain functional connectivity features of the brain functional network were extracted by principal component analysis algorithm and a 3-layer stacked denoising autoencoder; finally, feature selection and ensemble learning were carried out for the above three brain functional features, and a classification accuracy of 72% was obtained on multi-site data of ABIDE dataset. The experimental result illustrates that the proposed method can effectively improve the classification performance of ASD and NC. Compared with single-view learning, multi-view ensemble learning can mine various brain functional features of fMRI data from different perspectives and alleviate the problems caused by data heterogeneity. In addition, this study also employed leave-one-out cross validation to test the single-site data, and the results showed that the proposed method has strong generalization capability, in which the highest classification accuracy of 92.9% was obtained at the CMU site.


Intracellular taurine deficiency impairs cardiac contractility in rainbow trout (Oncorhynchus mykiss) without affecting aerobic performance.
Taurine is a non-proteinogenic sulfonic acid found in high concentrations inside vertebrate cardiomyocytes and its movement across the sarcolemmal membrane is critical for cell volume regulation. Taurine deficiency is rare in mammals, where it impairs cardiac contractility and leads to congestive heart failure. In fish, cardiac taurine levels vary substantially between species and can decrease by up to 60% in response to environmental change but its contribution to cardiac function is understudied. We addressed this gap in knowledge by generating a taurine-deficient rainbow trout (Oncorhynchus mykiss) model using a feed enriched with 3% β-alanine to inhibit cellular taurine uptake. Cardiac taurine was reduced by 17% after 4 weeks with no effect on growth or condition factor. Taurine deficiency did not affect routine or maximum rates of O2 consumption, aerobic scope, or critical swimming speed in whole animals but cardiac contractility was significantly impaired. In isometrically contracting ventricular strip preparations, the force-frequency and extracellular Ca2+-sensitivity relationships were both shifted downward and maximum pacing frequency was significantly lower in β-alanine fed trout. Cardiac taurine deficiency reduces sarcoplasmic reticular Ca2+-ATPase activity in mammals and our results are consistent with such an effect in rainbow trout. Our data indicate that intracellular taurine contributes to the regulation of cardiac contractility in rainbow trout. Aerobic performance was unaffected in β-alanine-fed animals, but further study is needed to determine if more significant natural reductions in taurine may constrain performance under certain environmental conditions.


Bifurcation lesion morphology and intravascular ultrasound assessment.
This repot reviews the angiographic and intravascular (IVUS) assessment of coronary bifurcation lesions. Overall, bifurcation lesion anatomy and morphology is critical for technical decision making, and a key factor for successful bifurcation PCI. Optimal viewing is essential for proper angiographic assessment, especially for evaluation of the degree of SB involvement. Current classifications based on the presence or absence of significant angiographic stenosis within the three segments of the bifurcation anatomy may not provide sufficient anatomic and morphologic information to guide technical decision making. Dedicated 2D bifurcation quantitative coronary angiography with segmental analysis of the bifurcation provides greater accuracy for quantification of the degree of stenosis in the PV and especially the SB ostium. IVUS assessment at preprocedure provides valuable information regarding vessel size, and plaque morphology and distribution (particularly in relation to the SB ostium) that may help select treatment strategy. At postprocedure, IVUS imaging evaluates stent apposition within the stented segment(s) and the appropriateness of stent expansion particularly at the SB ostium, what may impact long-term outcomes.


High dietary fat promotes visceral obesity and impaired endothelial function in female mice with systemic lupus erythematosus.
Inflammation contributes to metabolic and cardiovascular disease. Systemic lupus erythematosus (SLE) is a chronic autoimmune inflammatory disorder that predominantly affects young women. Cardiovascular disease is a major cause of mortality in patients with SLE. We recently reported that a model of SLE (female New Zealand Black/White F1 [NZBWF1] mice) developed characteristics of the metabolic syndrome. In the present study, we tested the hypothesis that high dietary fat with SLE accelerated development of cardiovascular risk factors such as central obesity and vascular dysfunction. Twenty-four-week-old female SLE mice (NZBWF1) were fed either a control diet (SLE, 10% kcal) or a high-fat (HF) diet (SLE + HF, 45% kcal) for a total of 14 weeks. Body weight was similar between SLE (42 [1] g, n = 5) and SLE + HF (45 [2] g, n = 6) mice, and weight gain was not different in the SLE + HF mice (+18.0 [3.0]%) compared with controls (+15.8 [3.6]%); food intake was not different (SLE, 2.2 [0.3] vs SLE + HF, 2.1 [0.2] g/24 hours). At the end of the experiment, 57% of the SLE + HF mice exhibited signs of albuminuria (>100 mg/dL) compared with only 20% of the control SLE mice. Endothelial-dependent relaxation in isolated carotid arteries was impaired in the SLE + HF group compared with that in the SLE group. Ovarian fat increased in SLE + HF mice (6.6 [0.5] g) compared with that in the control SLE mice (5.4 [0.1] g, P < 0.05), and liver weight decreased in SLE + HF (1.6 [0.1] g) mice compared with that in control mice (1.9 [0.1] g, P < 0.03). These data suggest that dietary fat accelerates renal injury and peripheral vascular dysfunction and promotes visceral obesity in a disease model with chronic inflammation.


Novel analogues of bradykinin conformationally restricted in the C-terminal part of the molecule.
In the present work, achiral non-coded amino acids, N-(Bzl)-Gly, X(1) or X(2) , were substituted at position 7 of the model B(2) receptor antagonist [D-Arg(0) , Hyp(3) , Thi(5, 8) , D-Phe(7) ]-BK. The N-terminal amino group of the analogues was either free or acylated with 1-Aca or Aaa. Biological activity of the compounds was assessed in the in vitro rat uterus test and the in vivo rat blood pressure test. The X(1) (7) substitution resulted in a decrease in antagonistic potency of the new peptide in both assays. The X(2) (7) and N-(Bzl)-Gly(7) substituted analogues showed weak agonistic properties in the rat uterus test. Interestingly, the latter compound exhibited dual activity in the pressor test, i.e. intrinsic vasodepressor action and at the same time a weak antagonistic effect. Acylation of the N-terminus enhanced antagonistic properties of the resulting peptides in the rat blood pressure test in the case of compounds containing X(1) or X(2) modification. Our studies provide new information about structure-activity relationship of the BK antagonists which may be helpful for designing more potent B(2) receptor blockers.


A MOF-on-MOF composite encapsulating sensitized Tb(III) as a built-in self-calibrating fluorescent platform for selective sensing of F ions.
The establishment of simple and sensitive detection methods for fluoride ion (F-) is of great importance for its effective prevention and control, and metal-organic framework (MOF) has attracted much attention for sensing applications due to its high surface areas and tunable structures. Herein, we successfully synthesized a fluorescent probe for ratiometric sensing of F- by encapsulating sensitized Tb3+ in a MOF-on-MOF material (UIO66/MOF801, with the formula of C48H28O32Zr6 and C24H2O32Zr6, respectively). We found that Tb3+@UIO66/MOF801 can be used as a built-in fluorescent probe for fluorescence-enhanced sensing of F-. Interestingly, the two fluorescence emission peaks of Tb3+@UIO66/MOF801 at 375 nm and 544 nm exhibit different fluorescence responses to F- under excitation at 300 nm. The 544 nm peak is sensitive to F-, while the 375 nm peak is insensitive to it. Photophysical analysis indicated that the photosensitive substance was formed, which promotes the absorption of 300 nm excitation light by the system. Self-calibrating fluorescent detection of F- was achieved due to the unequal energy transfer toward the two different emission centers. The detection limit of Tb3+@UIO66/MOF801 for F- was 4.029 μM, which is far lower than the WHO guideline for drinking water. Moreover, the ratiometric fluorescence strategy showed a high concentration tolerance of interference, because of its inner-reference effect. This work highlights the high potential of lanthanide ion encapsulated MOF-on-MOF as environmental sensors, and offers a scalable way for construction of the ratiometric fluorescence sensing systems.


Oscillations and bifurcation structure of reaction-diffusion model for cell polarity formation.
We investigate the oscillatory dynamics and bifurcation structure of a reaction-diffusion system with bistable nonlinearity and mass conservation, which was proposed by (Otsuji et al., PLoS Comp Biol 3:e108, 2007). The system is a useful model for understanding cell polarity formation. We show that this model exhibits four different spatiotemporal patterns including two types of oscillatory patterns, which can be regarded as cell polarity oscillations with the reversal and non-reversal of polarity, respectively. The trigger causing these patterns is a diffusion-driven (Turing-like) instability. Moreover, we investigate the effects of extracellular signals on the cell polarity oscillations.


Histochemical analysis of the masseter and temporalis muscles in macaca mulatta after mandibular advancement using rigid or nonrigid fixation.
This study compared the histochemical characteristics of the temporalis and masseter muscles in monkeys who underwent mandibular advancement with and without immobilization of the mandible. The results showed, when compared with controls, that the animals who underwent maxillomandibular fixation (MMF) following advancement of the mandible had significant atrophy in both temporalis and masseter muscles. In contrast, much less myoatrophy was noted in the animals who underwent rigid internal fixation and no MMF following advancement of the mandible.


Stable isotope labelling with amino acids in cell culture for human embryonic stem cell proteomic analysis.
The identification and quantitative measurements of proteins in human embryonic stem cells (hESC) is a fast growing interdisciplinary area with an enormous impact on understanding the biology of hESC and the mechanism controlling self-renewal and differentiation. Using a quantitative mass spectroscopic method of stable isotope labelling with amino acids during cell culture (SILAC), we are able to analyse differential expression of proteins from different cellular compartments and to identify intracellular signalling pathways involved in self-renewal and differentiation. In this chapter, we provide a detailed method for creating SILAC media suitable for use in hESC experiments, additionally we describe methods for the isolation of membrane fractions and cytosolic and nuclear/membrane fractions.


Complicated hydronephrosis of pregnancy.
The aggravated course during pregnancy of previously moderate hydronephrosis due to functional obstruction at the pelvi-ureteral junction is described. One patient underwent nephrectomy after parturition, because of severe hematuria, while another woman had a percutaneous nephrostomy in late pregnancy but could complete her pregnancy without further renal damage.


Intercropped Amygdalus persica and Pteris vittata applied with additives presents a safe utilization and remediation mode for arsenic-contaminated orchard soil.
Intercropping the arsenic (As) hyperaccumulator Pteris vittata with fruit trees can safely yield peaches in As-polluted orchards in South China. However, the soil As remediation effects and the related mechanisms of P. vittata intercropped with peach trees with additives in the north temperate zone have rarely been reported. A field experiment was conducted to systematically study the intercropping of peach (Amygdalus persica) with P. vittata with three additives [calcium magnesium phosphate (CMP), ammonium dihydrogen phosphate (ADP), and Stevia rebaudiana Bertoni residue (SR)] in a typical As-contaminated peach orchard surrounding a historical gold mine in Pinggu County, Beijing City. The results showed that compared with monoculture (PM) and intercropping without addition (LP), the remediation efficiency of P. vittata intercropping was significantly increased by 100.9 % (CMP) to 293.5 % (ADP). CMP and ADP mainly compete with available As (A-As) adsorbed to the surface of Fe-Al oxide through PO43-, while SR might activate A-As by enhancing dissolved organic carbon (DOC) in P. vittata rhizospheres. The photosynthetic rates (Gs) of intercropped P. vittata were significantly positively correlated with pinna As. The intercropping mode applied with the three additives did not obviously affect fruit quality, and the net profit of the intercropping mode (ADP) reached 415,800 yuan·ha-1·a-1. The As content in peaches was lower than the national standard in the intercropping systems. Comprehensive analysis showed that A. persica intercropped with P. vittata applied with ADP is better than other treatments in improving risk reduction and agricultural sustainability. In this study, a theoretical and practical basis is provided for the safe utilization and remediation of As-contaminated orchard soil in the north temperate zone.


Ventricular tachycardia in an adolescent with severe diabetic ketoacidosis.
Diabetic ketoacidosis (DKA) occurs frequently among children and adolescents with type 1 diabetes. Although a number of complications may occur during episodes of DKA, significant cardiac arrhythmias are uncommon. We present the case of an adolescent who presented with severe DKA and wide complex tachycardia that was unresponsive to multiple doses of adenosine and ultimately required synchronized cardioversion. This case reminds the clinician of the importance of cardiac monitoring in children with DKA, particularly in the setting of hyperkalemia.


Next generation sequencing-based multigene panel for high throughput detection of food-borne pathogens.
Contamination of food by chemicals or pathogenic bacteria may cause particular illnesses that are linked to food consumption, commonly referred to as foodborne diseases. Bacteria are present in/on various foods products, such as fruits, vegetables and ready-to-eat products. Bacteria that cause foodborne diseases are known as foodborne pathogens (FBPs). Accurate detection methods that are able to reveal the presence of FBPs in food matrices are in constant demand, in order to ensure safe foods with a minimal risk of causing foodborne diseases. Here, a multiplex PCR-based Illumina sequencing method for FBP detection in food matrices was developed. Starting from 25 bacterial targets and 49 selected PCR primer pairs, a primer collection called foodborne pathogen - panel (FPP) consisting of 12 oligonucleotide pairs was developed. The FPP allows a more rapid and reliable identification of FBPs compared to classical cultivation methods. Furthermore, FPP permits sensitive and specific FBP detection in about two days from food sample acquisition to bioinformatics-based identification. The FPP is able to simultaneously identify eight different bacterial pathogens, i.e. Listeria monocytogenes, Campylobacter jejuni, Campylobacter coli, Salmonella enterica subsp. enterica serovar enteritidis, Escherichia coli, Shigella sonnei, Staphylococcus aureus and Yersinia enterocolitica, in a given food matrix at a threshold contamination level of 101cell/g. Moreover, this novel detection method may represent an alternative and/or a complementary approach to PCR-based techniques, which are routinely used for FBP detection, and could be implemented in (parts of) the food chain as a quality check.


The molecular mechanisms and therapeutic potential of microRNA-7 in cancer.
Increasing evidence supports that microRNAs (miRNAs) play crucial roles in cancer through post-transcriptional gene silencing of their target genes, therefore, more and more effort has been devoted to develop miRNA-targeting therapeutics in cancer. MicroRNA-7 (miR-7) has been characterized as a potential tumor suppressor and regulates diverse fundamental biological processes of cancer cells including initiation, proliferation, migration, invasion, survival and death by targeting a number of oncogenic signaling pathways. This review examines evidence of the biological responses of miR-7 in cancer, with an emphasis on its regulation of the vital oncogenic signaling pathways. It also discusses the rationale, strategies and challenges of miR-7 as a potential therapeutic target for cancer. With the increasing understanding of molecular mechanisms of miR-7-mediated regulatory networks and the advancement of miRNA-based therapeutics, targeting miR-7 may be a potential and promising strategy for cancer therapy.


Cadmium-induced IL-6 and IL-8 expression and release from astrocytes are mediated by MAPK and NF-κB pathways.
Chronic exposure to cadmium has been linked to brain cancers, learning disabilities and memory deficits. Previous studies of cadmium toxicity in the central nervous system report cadmium induces oxidative stress in neurons and astrocytes. In the peripheral system, cadmium promotes interleukin-6 (IL-6) and IL-8 production and release. Elevation of IL-6 expression is linked to the pathogenesis of neurodegenerative diseases and astrogliosis. IL-8 plays a role in angiogenesis of gliomas and neurodegenerative diseases. Herein, the effects of non-toxic concentrations of cadmium on the production of IL-6 and IL-8 and the underlying mechanisms were investigated. U-87 MG human astrocytoma cells and primary human astrocytes were exposed to cadmium chloride. At 24h post-exposure to 1 and 10μM, levels of intracellular cadmium in U-87 MG cells were 11.89±3.59 and 53.08±7.59μg/g wet weight, respectively. These concentrations had minimal effects on cell morphology and viability. IL-6 and IL-8 mRNA levels and secretion increased in dose- and time-dependent manners post cadmium exposure. Acute exposure to cadmium increased phosphorylation of ERK1/2, p38 MAPK, and p65 NF-κB. Pretreatment with U0126-an inhibitor of MEK1 and MEK2 kinases-SB203580-a p38 MAPK inhibitor-and SC-514-an IKKβ inhibitor-suppressed cadmium-induced IL-8 expression and release. Upregulation of cadmium-induced IL-6 was inhibited by U0126 and SC-514, but not SB203580. On the other hand, SP600125-a JNK inhibitor-and celecoxib-a selective COX-2 inhibitor-had no effect on production of both cytokines. In conclusion, non-toxic concentrations of cadmium can stimulate IL-6 and IL-8 release through MAPK phosphorylation and NF-κB activation. Suppressing IL-6 and IL-8 production could be novel approaches to prevent cadmium-induced angiogenesis in gliomas and inflammation in the brain.


Routine detection of human rotavirus by latex agglutination: comparison with enzyme-linked immunosorbent assay, electron microscopy and polyacrylamide gel electrophoresis.
A commercially available latex agglutination test, RotaScreen (Mercia Diagnostics Ltd., West Byfleet, Surrey, U.K.) was evaluated for the detection of human rotaviruses in stool specimens. The results obtained were compared with those from 3 other routine assay systems used in this laboratory: enzyme-linked immunosorbent assay (ELISA), electron microscopy (EM) and polyacrylamide gel electrophoresis (PAGE) of viral ribonucleic acid. 400 stool samples were examined by the 4 assay systems under routine conditions. RotaScreen latex agglutination was found to be more sensitive than EM and PAGE, and highly specific for rotavirus antigens.


Power spectral analysis of two-channel EEG in very premature infants undergoing heat loss prevention.
To evaluate whether wearing a wool cap, a routine practice used to prevent heat loss in premature infants, affects interpretation of electroencephalogram spectral analysis. Eighteen premature infants (median gestational age 28 weeks, range 23-32) without neurological complications were randomized to two channel (C3, C4 referred to Cz) digital electroencephalogram recordings with (90 min) and without (90 min) wearing wool cap, at 4 days of life. Electroencephalogram was analyzed automatically by measurement of burst suppression ratio and asymmetry index and by Fast Fourier Transform to calculate total absolute spectral power; relative spectral power in the δ (0.5-3.5 Hz), θ (4-7.5 Hz), α (8-12.5 Hz), and β (13-30 Hz) frequency bands; spectral edge frequency; and mean dominant frequency. The use of wool cap had no effect on all electroencephalogram parameters considered. Gestational age showed an effect on relative spectral power of all considered bands, spectral edge frequency and mean dominant frequency, while no effect was seen on burst suppression ratio and asymmetry index. Neonates born at gestational weeks lower than 28 had significantly higher relative power in the δ band and lower relative power in the α and β bands. Heat loss prevention using wool cap does not affect interpretation of spectral electroencephalogram. Spectral values in our group of very premature infants without neurological complications correspond to normal data reported in the literature. Maturation changes consist of reduction of relative power of the δ band, spectral edge frequency and mean dominant frequency.


Reevaluation of the ambulatory pediatric patient whose blood culture is positive for Haemophilus influenzae type b.
We conducted a retrospective review of 60 patients with Haemophilus influenzae type b bacteremia initially treated as outpatients, to test the hypothesis that a subgroup of these patients is at low risk of continuing invasive infection and thus requires a less comprehensive reevaluation. These patients were 6% of the 975 patients with invasive H. influenzae type b infection identified by active surveillance in Dallas County, Texas, during a 6 1/2-year period. The clinical assessment of "ill" appearance and persistent fever (greater than or equal to 38.0 degrees C) on the return visit were the two most useful variables in identifying patients at risk of continuing infection. Among 25 (42%) patients who were considered "ill" (febrile or afebrile), 20 (80%) had continuing invasive infection and 14 (56%) had a subsequent culture that was positive for H. influenzae type b. Among 8 patients who were considered "well" but who remained febrile, 3 (38%) had continuing infection, including 1 (13%) patient with a second culture that was positive. By contrast, among 27 (45%) patients who were considered "well" and who were afebrile, continuing invasive infection was identified in only 2 (7%) patients, and all repeat cultures were negative. These results suggest that when patients return for reevaluation of H. influenzae type b bacteremia, clinical assessment by an experienced physician, together with the febrile status of the patient, can identify those at high versus low risk of continuing invasive infection and thus guide the extent of diagnostic reevaluation.


Stereoselective synthesis of novel 2'-(S)-CCG-IV analogues as potent NMDA receptor agonists.
We developed a versatile stereoselective route for the synthesis of new 2'-(S)-CCG-IV analogues. The route allows for late stage diversification and thereby provides access to a great variety of conformationally restricted cyclopropyl glutamate analogues. A selection of the 2'-(S)-CCG-IV analogues were evaluated using two-electrode voltage-clamp electrophysiology at recombinant GluN1/GluN2A-D receptors, demonstrating that agonists can be developed with GluN2 subunit-dependent potency and agonist efficacy. We also describe a crystal structure of the GluN2A agonist binding domain in complex with 2'-butyl-(S)-CCG-IV that determines the position of 2'-substituents in (S)-CCG-IV agonists in the glutamate binding site and provides further insight to the structural determinants of their agonist efficacy. The stereoselective synthesis described here enables versatile and straight-forward modifications to diverse analogues of interest for the development of potent subtype-specific NMDA receptor agonists and other applications.


Development and optimization of an ultra-fast microextraction followed by HPLC-UV of tetracycline residues in milk products.
The present study developed a new ultra-fast microextraction (within 8 min) with acetonitrile followed by gradient reversed-phase HPLC-UV method to determinate six tetracyclines simultaneously in various milk products with HPLC analysis time of 9 min. Chromatographic separations were achieved with a phenomenex™ Synergi 4 µm Fusion-RP (150 × 4.60 mm) column at 220 nm, using acetonitrile/KH2PO4 buffer (4 mmol·L-1, pH 2.5) as mobile phase. The HPLC method showed excellent peak resolutions (3.3-8.8) and peak symmetry (0.99-1.11) inspite of short analysis time. The extraction rates from milk products showed consistently very good values over all tetracyclines (milk 80.58 ± 5.39 %, yoghurt 82.61 ± 5.63 %, cream cheese 80.13 ± 6.32 %, buttermilk 81.07 ± 6.49 %, kefir 79.69 ± 6.51 %, skyr 78.12 ± 5.22 %, quark 65.37 ± 4.72 %). The optimized method was found to be specific, reproducible, robust. This study combines for the first time a fast, cheap quantification of six tetracyclines via HPLC-UV with a reliable microextraction applicable to various milk products using only standard laboratory equipment.


Nature of Alkali- and Coinage-Metal Bonds versus Hydrogen Bonds.
We have quantum chemically studied the structure and nature of alkali- and coinage-metal bonds (M-bonds) versus that of hydrogen bonds between A-M and B- in archetypal [A-M⋅⋅⋅B]- model systems (A, B=F, Cl and M=H, Li, Na, Cu, Ag, Au), using relativistic density functional theory at ZORA-BP86-D3/TZ2P. We find that coinage-metal bonds are stronger than alkali-metal bonds which are stronger than the corresponding hydrogen bonds. Our main purpose is to understand how and why the structure, stability and nature of such bonds are affected if the monovalent central atom H of hydrogen bonds is replaced by an isoelectronic alkali- or coinage-metal atom. To this end, we have analyzed the bonds between A-M and B- using the activation strain model, quantitative Kohn-Sham molecular orbital (MO) theory, energy decomposition analysis (EDA), and Voronoi deformation density (VDD) analysis of the charge distribution.


Contribution of dual fluorescein and indocyanine green angiography to the appraisal of posterior involvement in birdshot retinochoroiditis and Vogt-Koyanagi-Harada disease.
To assess the levels of retinal and choroidal involvement in initial-onset birdshot retinochoroiditis (BRC) and Vogt-Koyanagi-Harada (VKH) disease, two stromal choroiditis entities. This retrospective study included patients diagnosed with BRC and VKH, seen during initial-onset disease at the Centre for Ophthalmic Specialized Care, Lausanne, Switzerland. Angiographic signs were quantified, using an established dual fluorescein angiography (FA) and indocyanine green angiography (ICGA) scoring system for uveitis, and the FA/ICGA score ratios were compared between diseases. Among 1793 patients with uveitis seen from 1995 to 2015, 7 newly diagnosed BRC patients and 4 patients with newly diagnosed VKH disease had sufficient data for study inclusion. Patients with BRC and VKH at initial onset had mean FA angiographic scores of 16.91 ± 3.42 and 4.06 ± 1.87; mean ICGA angiographic scores of 21.34 ± 3.49 and 25.75 ± 3.88; and mean FA/ICGA ratios of 0.79 ± 0.21 and 0.16 ± 0.09, respectively. This study showed the differential involvements of the retina and choroid in BRC and VKH. The choroid was preponderantly involved in both diseases; thus, ICGA is essential for disease assessment and follow-up. However, these diseases also differed substantially. The origin of inflammation was primarily in the choroid in VKH and in both the choroid and retina in BRC. We recommend dual FA and ICGA for evaluating posterior uveitis, when choroiditis is suspected.


Metabolomic networks in plants: Transitions from pattern recognition to biological interpretation.
Nowadays techniques for non-targeted metabolite profiling allow for the generation of huge amounts of relevant data essential for the construction of dynamic metabolomic networks. Thus, metabolomics, besides transcriptomics or proteomics, provides a major tool for the characterization of postgenomic processes. In this work, we introduce comparative correlation analysis as a complementary approach to characterize the physiological states of various organs of diverse plant species with focus on specific participation of metabolites in different reaction networks. The correlations observed are induced by diminutive fluctuations in environmental conditions, which propagate through the system and induce specific patterns depending on the genomic background. In order to examine this hypothesis, numeric examples of such fluctuations are computed and compared with experimentally obtained metabolite data.


Clinical outcomes after 4.5 years of eliglustat therapy for Gaucher disease type 1: Phase 3 ENGAGE trial final results.
Eliglustat, an oral substrate reduction therapy, is approved for eligible adults with Gaucher disease type 1. In the Phase 3 ENGAGE trial of previously untreated adults with Gaucher disease type 1, eliglustat-treated patients had statistically significant improvements in organ volumes and hematologic parameters compared with placebo in the 9-month primary analysis. We report final outcomes by time on eliglustat among all patients who participated in the ENGAGE trial and extension. No patient deteriorated clinically or withdrew due to adverse events; 39/40 patients entered the open-label extension period and 34/40 (85%) remained in the trial until completion or switching to commercial eliglustat after its approval (2.3-6 years). Clinically meaningful improvements in Gaucher disease manifestations were seen in all patients concomitant with reductions in pathological lipid substrate levels (glucosylceramide and glucosylsphingosine). Among patients with 4.5 years of eliglustat exposure, mean spleen volume decreased by 66% (from 17.1 to 5.8 multiples of normal [MN], n = 13), mean liver volume decreased by 23% (from 1.5 to 1.1 MN, n = 13), mean hemoglobin increased 1.4 g/dl (from 11.9 to 13.4 g/dl, n = 12), mean platelet count increased by 87% (from 67.6 to 122.6 × 109 /L, n = 12), median chitotriosidase decreased by 82% (from 13 394 to 2312 nmol/h/ml, n = 11), median glucosylceramide decreased by 79% (from 11.5 to 2.4 μg/ml, n = 11), median glucosylsphingosine decreased by 84% (from 518.5 to 72.1 ng/ml, n = 10), and mean spine T-score increased from -1.07 (osteopenia) to -0.53 (normal) (n = 9). The magnitude of improvement in Gaucher disease manifestations and biomarkers over time was similar among the full trial cohort. Eliglustat was well-tolerated and led to clinically significant improvements in previously untreated patients with Gaucher disease type 1 during 4.5 years of treatment.


Comparison of long-term outcomes between open and laparoscopic Thal fundoplication in children.
In recent years laparoscopic fundoplication is increasingly performed in pediatric surgery. The aim of this study was to compare the long-term outcomes between open and laparoscopic Thal fundoplication in children. This retrospective study includes children who underwent a Thal fundoplication between 3/1997 and 7/2009. The minimum follow-up time to enter the study was 2 years; the overall median follow-up was 77 months (range, 29-176 months). A total of 101 patients were included, of which 47 underwent an open and 54 a laparoscopic Thal. Intraoperative problems, early postoperative complications, time to establish enteral feeds and length of stay did not differ among both groups. The mean duration of surgery was significantly less in the open group (OPG) (108.0 (± 7.72) versus 144.1 (± 6.36) minutes; p=0.001) and this was mainly attributed to patients with neurological problems. Severe dysphagia requiring endoscopy was observed in 10 patients, but this did not differ significantly between groups (n=2 in the OPG vs. n=8 in the laparoscopic group (LAPG); p=0.10). Overall 12 patients (11.9%) (6 in each group) required a redo-fundoplication after a median of 18.7 months (range, 6-36 months). In the whole study group, 80 patients (79.2%) were classified as having surgical results being excellent, good or satisfactory and this did not differ significantly between groups. In the long-term open and laparoscopic Thal fundoplication have similarly good outcomes. The laparoscopic approach can be considered as an alternative, however there is not a clear superiority compared with the open counterpart.


Coconut Mesocarp-Based Lignocellulosic Waste as a Substrate for Cellulase Production from High Promising Multienzyme-Producing Bacillus amyloliquefaciens FW2 without Pretreatments.
Facing the crucial issue of high cost in cellulase production from commercial celluloses, inexpensive lignocellulosic materials from agricultural wastes have been attractive. Therefore, several studies have focused on increasing the efficiency of cellulase production by potential microorganisms capable of secreting a high and diversified amount of enzymes using agricultural waste as valuable substrates. Especially, extremophilic bacteria play an important role in biorefinery due to their high value catalytic enzymes that are active even under harsh environmental conditions. Therefore, in this study, we aim to investigate the ability to produce cellulase from coconut-mesocarp of the potential bacterial strain FW2 that was isolated from kitchen food waste in South Korea. This strain was tolerant in a wide range of temperature (-6-75 °C, pH range (4.5-12)) and at high salt concentration up to 35% NaCl. The molecular weight of the purified cellulase produced from strain FW2 was estimated to be 55 kDa. Optimal conditions for the enzyme activity using commercial substrates were found to be 40-50 °C, pH 7.0-7.5, and 0-10% NaCl observed in 920 U/mL of CMCase, 1300 U/mL of Avicelase, and 150 U/mL of FPase. It was achieved in 650 U/mL, 720 U/mL, and 140 U/mL of CMCase, Avicelase, and FPase using coconut-mesocarp, respectively. The results revealed that enzyme production by strain FW2 may have significant commercial values for industry, argo-waste treatment, and other potential applications.


Discovery of a rapidly evolving yeast defense factor, KTD1, against the secreted killer toxin K28.
Secreted protein toxins are widely used weapons in conflicts between organisms. Elucidating how organisms genetically adapt to defend themselves against these toxins is fundamental to understanding the coevolutionary dynamics of competing organisms. Within yeast communities, "killer" toxins are secreted to kill nearby sensitive yeast, providing a fitness advantage in competitive growth environments. Natural yeast isolates vary in their sensitivity to these toxins, but to date, no polymorphic genetic factors contributing to defense have been identified. We investigated the variation in resistance to the killer toxin K28 across diverse natural isolates of the Saccharomyces cerevisiae population. Using large-scale linkage mapping, we discovered a novel defense factor, which we named KTD1. We identified many KTD1 alleles, which provided different levels of K28 resistance. KTD1 is a member of the DUP240 gene family of unknown function, which is rapidly evolving in a region spanning its two encoded transmembrane helices. We found that this domain is critical to KTD1's protective ability. Our findings implicate KTD1 as a key polymorphic factor in the defense against K28 toxin.


Sterolomics: State of the art, developments, limitations and challenges.
Sterolomics can be thought of as the quantitative determination of the entire complement of molecules based on the cyclopentanoperhydrophenanthrene skeleton in a system. Mass spectrometry is the dominant analytical technology employed. In this article we highlight some pitfalls in analysis, data interpretation and annotation. We give our opinion on how some of these pitfalls can best be avoided. This article is part of a Special Issue entitled: BBALIP_Lipidomics Opinion Articles edited by Sepp Kohlwein.


Radon risk mapping: A new geostatistical method based on Lorenz Curve and Gini index.
In confined spaces such as living environments and workplaces, the concentration levels of radon (Rn222) can be very high as compared to the external environment. Since Rn has been classified as the second leading cause of lung cancer after cigarette smoking, to apply efficient locally based risk reduction actions, dense maps of indoor radon concentration are needed. These maps would provide information about the areas prone to high radon concentrations and therefore more dangerous to human health. The soil is the primary source of the Rn, hence the risk assessment and reduction for the radon exposure cannot disregard the identification of the local geology. In this regard, we propose an innovative method, based on the Gini index computation, for the realization of interpolated maps (kriging) to describe the distribution of concentration of Rn. To validate the method, a tool that simulates sets of radon concentrations is used, whose variability is, to the first order, controlled by a priori imposed different lithologies. A systematic comparison is made between the results achieved by means of a classically used geostatistical method and the proposed Gini-based tool. We show how, by using this latter tool, the kriging solutions appear to be more robust to resolve the different geogenic radon sources independently from the number of the available measurements.


Nanoporous SiC: a candidate semi-permeable material for biomedical applications.
We have fabricated free-standing SiC nanoporous membranes in both p -type and n -type material. We showed that these membranes will permit the diffusion of proteins up to 29000 Daltons, while excluding larger proteins. By using radioactively labeled albumin, we also show that porous SiC has very low protein adsorption, comparable to the best commercially available polymer nanoporous membrane.


X-ray crystallographic analysis of the antiferromagnetic low-temperature phase of galvinoxyl: investigating magnetic duality in organic radicals.
Galvinoxyl, as one of the most extensively studied organic stable free radicals, exhibits a notable phase transition from a high-temperature (HT) phase with a ferromagnetic (FM) intermolecular interaction to a low-temperature (LT) phase with an antiferromagnetic (AFM) coupling at 85 K. Despite significant research efforts, the crystal structure of the AFM LT phase has remained elusive. This study successfully elucidates the crystal structure of the LT phase, which belongs to the P1̄ space group. The crystal structure of the LT phase is found to consist of a distorted dimer, wherein the distortion arises from the formation of short intermolecular distances between anti-node carbons in the singly-occupied molecular orbital (SOMO). Starting from the structure of the LT phase, wave function calculations show that the AFM coupling 2J/kB varies significantly from -1069 K to -54 K due to a parallel shift of the molecular planes within the dimer.


Integration of iron oxide nanoparticles and polyaspartamide biopolymer for MRI image contrast enhancement and an efficient drug-delivery system in cancer therapy.
We integrate superparamagnetic iron oxide nanoparticles with polyaspartamide (PA) biopolymer to form a biological construct that functions as a tracking, targeting and drug-delivery system for cancer diagnosis and therapy. Iron oxide nanoparticles with uniformly distributed average spherical diameters of around 10 nm and superparamagnetic characteristics play a key role in increasing the transverse 1/T 2 relaxation rate or darkening the T 2-weighted MR image for cancer diagnosis using MRI. In in vitro MRI testing on cancer cells, the MR images of samples with the bio-constructshow a much clearer contrast effect than those of controls. The PA biopolymer plays an essential role in enhancing the hydrophilicity and biocompatibility of the bio-construct. In addition, as a multifunctional polymer, PA is conjugated with biotin and doxorubicin (Dox) functional groups to enhance targeting and impairment of cancer cells. In in vivo testing on cancer tumors, injection with the bio-construct decreased the magnitude of cancer tumor volume growth by three times compared with that of uninjected controls. The physicochemical characteristics of the bio-construct and the roles of biotin and Dox functional groups are examined and discussed in detail.


Enhancing the flow field in parallel spiral-flow column photobioreactor to improve CO2 fixation with Spirulina sp.
A parallel spiral-flow column photobioreactor (PSCP) composed of eight spiral-flow columns, and two pipe headers was designed for scale-up cultivation of microalgae to capture CO2. To solve the disturbance of spiral flow fields among parallel columns, computational fluid dynamics (CFD) simulation was used to optimize the main structural parameters, such as the number and the height of microalgae solution outlet (MSO), to improve flow field structure and enhance the cells' light/dark cycle. The horizontal velocity in the direction of optical path and the turbulent kinetic energy (TKE) reached the peak values of 0.214 m/s and 5.28 m2/s2 when MSO number was four and MSO height was 1.05 m. Meanwhile, the disturbance of the spiral flow field among parallel columns are minimum, and microalgae light/dark cycle frequency was 33.3% higher than that of conventional bubble column photobioreactor. Therefore, the biomass yield and CO2 fixation rate of microalgae increased by 81.5% and 100.5%, respectively.


Diabetic ketoacidosis in patients with SARS-CoV-2: a systematic review and meta-analysis.
One possible reason for increased mortality due to SARS-CoV-2 in patients with diabetes is from the complication of diabetic ketoacidosis (DKA). To re-evaluate the association of SARS-CoV-2 and development of DKA and analyse the demographic and biochemical parameters and the clinical outcomes in COVID-19 patients with DKA. A systematic review and meta-analysis. Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement was followed. Electronic databases (Proquest, Medline, Embase, Pubmed, CINAHL, Wiley online library, Scopus and Nature) were searched from 1 December 2019 to 30 June 2021 in the English language using the following keywords alone or in combination: COVID-19 OR SARS-CoV-2 AND diabetic ketoacidosis OR DKA OR ketosis OR ketonemia OR hyperglycaemic emergency OR hyperglycaemic crisis. We included studies in adults and children of all ages in all healthcare settings. Binary logistic regression model was used to explore the effect of various demographic and biochemical parameters variables on patient's final treatment outcome (survival or death). Of the 484 papers that were identified, 68 articles were included in the systematic review and meta-analysis (54 case report, 10 case series, and 4 cohort studies). Studies involving 639 DKA patients with confirmed SARS-CoV-2 [46 (7.2%) were children and 334 (52.3%) were adults] were analyzed. The median or mean patient age ranged from < 1 years to 66 years across studies. Most of the patients (n = 309, 48.3%) had pre-existing type 2 diabetes mellitus. The majority of the patients were male (n = 373, 58.4%) and belonged to Hispanic (n = 156, 24.4%) and black (n = 98, 15.3%) ethnicity. The median random blood glucose level, HbA1c, pH, bicarbonate, and anion gap in all included patients at presentation were 507 mg/dl [IQR 399-638 mg/dl], 11.4% [IQR 9.9-13.5%], 7.16 [IQR 7.00-7.22], 10 mmol/l [IQR 6.9-13 mmol/l], and 24.5 mEq/l [18-29.2 mEq/l]; respectively. Mortality rate was [63/243, 25.9%], with a majority of death in patients of Hispanic ethnicity (n = 17, 27%; p = 0.001). The odd ratios of death were significantly high in patients with pre-existing diabetes mellitus type 2 [OR 5.24, 95% CI 2.07-15.19; p = 0.001], old age (≥ 60 years) [OR 3.29, 95% CI 1.38-7.91; p = 0.007], and male gender [OR 2.61, 95% CI 1.37-5.17; p = 0.004] compared to those who survived. DKA is not uncommon in SARS-CoV-2 patients with diabetes mellitus and results in a mortality rate of 25.9%. Mortality key determinants in DKA patients with SARS-CoV-2 infection are individuals with pre-existing diabetes mellitus type 2, older age [≥ 60 years old], male gender, BMI ≥ 30, blood glucose level > 1000 mg/dl, and anion gap ≥ 30 mEq/l.


Cancer genetics and genomics of human FOX family genes.
Forkhead-box (FOX) family proteins, involved in cell growth and differentiation as well as embryogenesis and longevity, are DNA-binding proteins regulating transcription and DNA repair. The focus of this review is on the mechanisms of FOX-related human carcinogenesis. FOXA1 is overexpressed as a result of gene amplification in lung cancer, esophageal cancer, ER-positive breast cancer and anaplastic thyroid cancer and is point-mutated in prostate cancer. FOXA1 overexpression in breast cancer and prostate cancer is associated with good or poor prognosis, respectively. Single nucleotide polymorphism (SNP) within the 5'-UTR of the FOXE1 (TTF2) gene is associated with thyroid cancer risk. FOXF1 overexpression in breast cancer is associated with epithelial-to-mesenchymal transition (EMT). FOXM1 is overexpressed owing to gene amplification in basal-type breast cancer and diffuse large B-cell lymphoma (DLBCL), and it is transcriptionally upregulated owing to Hedgehog-GLI, hypoxia-HIF1α or YAP-TEAD signaling activation. FOXM1 overexpression leads to malignant phenotypes by directly upregulating CCNB1, AURKB, MYC and SKP2 and indirectly upregulating ZEB1 and ZEB2 via miR-200b downregulation. Tumor suppressor functions of FOXO transcription factors are lost in cancer cells as a result of chromosomal translocation, deletion, miRNA-mediated repression, AKT-mediated cytoplasmic sequestration or ubiquitination-mediated proteasomal degradation. FOXP1 is upregulated as a result of gene fusion or amplification in DLBCL and MALT lymphoma and also repression of miRNAs, such as miR-1, miR-34a and miR-504. FOXP1 overexpression is associated with poor prognosis in DLBCL, gastric MALT lymphoma and hepatocellular carcinoma but with good prognosis in breast cancer. In neuroblastoma, the entire coding region of the FOXR1 (FOXN5) gene is fused to the MLL or the PAFAH1B gene owing to interstitial deletions. FOXR1 fusion genes function as oncogenes that repress transcription of FOXO target genes. Whole-genome sequencing data from tens of thousands of human cancers will uncover the mutational landscape of FOX family genes themselves as well as FOX-binding sites, which will be ultimately applied for cancer diagnostics, prognostics, and therapeutics.


Surgical procedure prediction using medical ontological information.
Predicting the duration of surgical procedures is an important step in scheduling operating rooms. Many factors have been shown to influence the duration of a procedure, in this research we aim to use medical ontological information to improve the predictions. This paper presents two methods for incorporating the medical information about a surgical procedure into the prediction of the duration of the procedure. The first method uses the Systematised Nomenclature of Medicine Clinical Terms to relate different procedures to each other. The second uses simple text fragments. The relationships between types of procedures are included in a regression model for the procedure duration. These methods are applied to data from New Zealand healthcare facilities and the accuracy of the estimations of the durations is compared. In addition a simulation of scheduling the procedures in an operating room is performed. It is shown that both of the methods provide an improvement in the prediction of procedure durations. When compared to a traditional categorical encoding, the ontological information provides an improvement in the continuous ranked probability scores of the prediction of procedure durations from 18.4 min to 17.1 min, and from 25.3 to 21.3 min for types of procedures that are not performed very often. Different methods for encoding medical ontological information in surgery procedure duration predictions are presented, and show an improvement over traditional models. The improvement in duration prediction is shown to improve the efficiency of scheduling in a simple simulation.


Characterization of cognitive function in survivors of diffuse gliomas using resting-state functional MRI (rs-fMRI).
As treatments for diffuse gliomas have advanced, survival for patients with gliomas has also increased. However, there remains limited knowledge on the relationships between brain connectivity and the lasting changes to cognitive function that glioma survivors often experience long after completing treatment. This resting-state functional magnetic resonance imaging (rs-fMRI) study explored functional connectivity (FC) alterations associated with cognitive function in survivors of gliomas. In this pilot study, 22 patients (mean age 43.8 ± 11.9) with diffuse gliomas who completed treatment within the past 10 years were evaluated using rs-fMRI and neuropsychological measures. Novel rs-fMRI analysis methods were used to account for missing brain in the resection cavity. FC relationships were assessed between cognitively impaired and non-impaired glioma patients, along with self-reported cognitive impairment, non-work daily functioning, and time with surgery. In the cognitively non-impaired patients, FC was stronger in the medial prefrontal cortex, rostral prefrontal cortex, and intraparietal sulcus compared to the impaired survivors. When examining non-work daily functioning, a positive correlation with FC was observed between the accumbens and the intracalcarine cortices, while a negative correlation with FC was observed between the parietal operculum cortex and the cerebellum. Additionally, worse self-reported cognitive impairment and worse non-work daily functioning were associated with increased FC between regions involved in cognition and sensorimotor processing. These preliminary findings suggest that neural correlates for cognitive and daily functioning in glioma patients can be revealed using rs-fMRI. Resting-state network alterations may serve as a biomarker for patients' cognition and functioning.


Age-dependent depressor effect of L-dopa in dogs with inhibition of extracerebral dopa decarboxylase.
The results of this study demonstrate that a carbidopa-L-dopa combination has a greater depressor effect in puppies than in adult dogs. These effects are believed to be due to the action of a metabolite of L-dopa within the CNS. These data should alert clinicians to the possibility that CNS cardiovascular control centers, which utilize catecholamines as neuromediators, may be more responsive to pharmacological stimulation in the young than in the adult.


Location of immunoglobulins in the skin of cattle and sheep.
The pattern of distribution of immunoglobulins (Ig) found within the skin of cattle and sheep was the same in both species. IgA was detected mainly in the sweat gland duct and in the intercellular spaces of the living epidermis. IgG was also located at the latter site but was found in greater amounts throughout the dermis and in the endothelium of the blood vessels. IgM was primarily located at the basement membrane of the epithelial structures, in the hair papilla and in the walls of the blood vessels. Lower concentrations of IgM were also detected in the upper dermis.


Achievable outcomes in the management of proximal cholangiocarcinoma: an update prepared using "evidence-based practice" techniques.
The purpose of this study is to evaluate recently reported outcomes for treatment options for proximal cholangiocarcinoma (CCA). Standard evidence based practice techniques were used to formulate a question, search, appraise and evaluate the retrieved literature. Our question was "In patients with CCA, how do stenting alone, stenting in addition to brachytherapy (BT) or photodynamic therapy (PDT), resection and orthotopic liver transplantation with neoadjuvant chemoradiation (OLT) compare for long-term survival? Level 1b survival data was available for stenting alone (179 days), BT and metal stenting (388 days) and PDT with plastic stenting (493 days) and no survival difference was evident with metal vs. plastic stenting or unilateral vs. bilateral stenting. Five year survival data (level 3) was available for OLT (80%), formally curative trisegmentectomy with or without portal vein resection (72% and 52%) and hepatectomy (18%-23%). All patients with proximal CCA should be reviewed by a multidisciplinary team to determine appropriate treatment. For unresectable CCA, patients should be assessed for OLT with neoadjuvant chemoradiation, while those who are unsuitable would appear to have the longest survival with PDT. Extended resection in operable candidates may improve survival over right or left hepatectomy but increased perioperative mortality is a consideration.


PUVA treatment of psoriasis in the United Kingdom.
Information concerning the operation of 134 PUVA units in 96 dermatology centres in the U.K. was collected by a postal questionnaire. The most common protocol was to give PUVA three times weekly in the treatment of psoriasis, and to choose the initial dose of UVA according to skin type. However, the initial UVA dose encompassed a range of five or more within each skin type.


Photoreactivity of 5-geranoxypsoralen and lack of photoreaction with DNA.
5-Geranoxypsoralen, commonly called bergamottin, a major furocoumarin contained in bergamot oil, is reported in vitro as a highly photoreactive psoralen. In ethanol, it exhibits quite a high triplet state quantum yield (approximately 0.37). The triplet state is involved in subsequent photochemistry which depends on the initial concentration and on the presence of oxygen. In contrast to most psoralens, absorption and fluorescence data suggest that 5-geranoxypsoralen does not interact with DNA in the dark. No UVA-induced interstrand cross-links in DNA were shown.


A robust mucin-containing poly(vinyl alcohol) hydrogel model for the in vitro characterization of mucoadhesion of solid dosage forms.
Mucoadhesion testing at macroscopic scale needs a robust, convenient in vitro method as ex vivo methods suffer from poor reproducibility and ethical problems. Here we synthesized mucin-free poly(vinyl alcohol) (PVA) and mucin-containing PVA hydrogel substrates (Muc/PVA) to measure adhesion of polymer tablets. Freezing-thawing method was used for gelation to avoid chemical cross-linking and to preserve the functionality of mucin. The adhesion of first generation mucoadhesive polymers, poly(acrylic acid) (PAA) and hydroxypropylmethylcellulose (HPMC) was tested with outstanding reproducibility on individual batches of hydrogels and qualitative agreement with ex vivo literature data. Negatively charged PAA was less adhesive on Muc/PVA surface than on mucin-free PVA whereas HPMC as a neutral polymer displayed similar adhesion strength on both surfaces. Chitosan as a positively charged polymer showed enhanced adhesion on Muc/PVA substrate compared to mucin-free PVA. These results are corroborated by turbidimetric titration which indicated attractive electrostatic interactions between chitosan and mucin in contrast to the lack of attractive interactions for PAA and HPMC. These results prove the role of electronic theory in macroscopic mucoadhesion.


Hormonal control of fat cells adenylate cyclase.
Fat cells were preincubated for 2 h in the presence and absence of growth hormone (GH) and Dexamethasone (Dex) before the addition of increasing concentrations of either epinephrine, theophylline or glucagon and final incubation of the cells for an additional 5 minutes. GH and Dex increased by 85%, 28% and 72%, respectively, the cAMP levels reached in the sole presence of 10(-5)M epinephrine, 10(-2)M theophylline or 5 X 10(-5)M glucagon. An adenylate cyclase particulate preparation shows that epinephrine decreases Km from 2mM to 0.6 MM and increases Vmax and the strength of interaction value (n) from 0.91 to 1.75.


Redesigning the T-probe for mass spectrometry analysis of online lysis of non-adherent single cells.
Single cell mass spectrometry (SCMS) allows for molecular analysis of individual cells while avoiding the inevitable drawbacks of using cell lysate prepared from populations of cells. Based on our previous design of the T-probe, a microscale sampling and ionization device for SCMS analysis, we further developed the device to perform online, and real time lysis of non-adherent live single cells for mass spectrometry (MS) analysis at ambient conditions. This redesigned T-probe includes three parts: a sampling probe with a small tip to withdraw a whole cell, a solvent-providing capillary to deliver lysis solution (i.e., acetonitrile), and a nano-ESI emitter in which rapid cell lysis and ionization occur followed by MS analysis. These three components are embedded between two polycarbonate slides and are jointed through a T-junction to form an integrated device. Colon cancer cells (HCT-116) under control and treatment (using anticancer drug irinotecan) conditions were analyzed. We detected a variety of intracellular species, and structural identification of selected ions was conducted using tandem MS (MS2). We further conducted statistical analysis (e.g., PLS-DA and t-test) to gain biological insights of cellular metabolism. Our results indicate that the influence of anticancer drugs on cellular metabolism of live non-adherent cells can be obtained using the SCMS experiments combined with statistical data analysis.


Transcriptional regulation by a RecQ helicase.
RecQ helicases participate in a variety of DNA metabolic processes through their multiple biochemical activities. In vitro characterization and cellular studies have suggested that RECQ1 (also known as RECQL or RECQL1) performs its diverse functions through specific interactions with DNA and protein partners. We have taken an unbiased approach to determine the contribution of RECQ1 in genome maintenance and as a putative susceptibility factor in breast cancer. Here, we provide methodology to map the genome-wide binding sites of RECQ1 together with the profiling of RECQ1-dependent transcriptome to investigate its role in gene regulation. The described approach will be helpful to develop a mechanistic framework for elucidating critical functions of RECQ1 and other RecQ homologs in distinct chromatin and biological contexts.


In Vitro Selection of Thioether-Closed Macrocyclic Peptide Ligands by Means of the RaPID System.
The Random nonstandard Peptides Integrated Discovery (RaPID) system enables efficient screening of macrocyclic peptides with high affinities against target molecules. Random peptide libraries are prepared by in vitro translation using the Flexible In vitro Translation (FIT) system, which allows for incorporation of diverse nonproteinogenic amino acids into peptides by genetic code reprogramming. By introducing an N-chloroacetyl amino acid at the N-terminus and a Cys at the downstream, macrocyclic peptide libraries can be readily generated via posttranslational thioether formation. Here, we describe how to prepare a thioether-closed macrocyclic peptide library, and its application to the RaPID screening.


[Navigated focal retinal laser therapy using the NAVILAS® system for diabetic macula edema].
The aim of this study was to investigate the accuracy of a navigated laser photocoagulator in clinically significant macular edema (CSME). Focal laser treatment for diabetic macular edema (DME) in 36 patients was digitally planned on fundus images and performed with navigation using NAVILAS® (OD-OS, Teltow, Germany). Treatment intensity was controlled visually during treatment so the laser spots applied were barely directly visible after treatment. Using color images (CI) and optical coherence tomography (OCT) 4,137 laser spots (mean 115 per eye) were analyzed at 1 month follow-up and accuracy of spot placement was determined. In total 79% of laser spots were visible on CI of which 96% were within 100 µm of the planned target position. On an intention-to-treat (ITT) basis, 76% of the laser spots were placed and visible within the 100 µm target and OCT confirmed that laser effects were limited to the outer retina. The mean time for focal treatment was < 7 min (±3 min). After NAVILAS treatment for DME a high percentage of laser effects could be visualized on post-treatment color images and the location showed high concordance with the preplanning target.