Determinants of health insurance status for children of Latino immigrant and other US farm workers: findings from the National Agricultural Workers Survey.
To characterize the health insurance status of farm workers' children, an understudied topic. A population-based multistage survey. Employer-based study conducted in the continental United States from 2000 to 2002. A total of 3136 parents with children younger than 18 years and no children residing outside of the United States who participated in the US Department of Labor's National Agricultural Workers Survey, which is administered to a national probability sample of US farm workers. Children's parent-reported health insurance status. Of the farm workers who were parents, 87% were Latino, 81% were foreign born, 15% were migrant workers, 55% had less than a sixth-grade education, and 68% reported little to no English language proficiency. Thirty-two percent of all farm-worker parents, including 45% of migrant-worker parents, reported that their children were uninsured. In multivariate analyses, older parental age (odds ratio [OR] for parents aged 30-39 years, 1.71; 95% confidence interval [CI], 1.16-2.50; OR for parents aged >/=40 years, 3.07; 95% CI, 1.99-4.74), less parental education (OR, 1.52; 95% CI, 1.09-2.10), less time in the United States (OR, 0.68; 95% CI, 0.56-0.91 per 10 years in the United States), being a migrant worker (OR, 1.96; 95% CI, 1.31-2.93), and living in the Southeast (OR, 3.17; 95% CI, 2.00-5.02) or Southwest (OR, 3.91; 95% CI, 2.32-6.57) were significantly associated with having uninsured children. Farm workers' children were uninsured at roughly 3 times the rate of all other children and almost twice the rate of those at or near the federal poverty level. Programs aimed at extending insurance coverage for children should consider the unique social barriers that characterize this vulnerable population of US children. Moreover, there is significant regional variation that may reflect varying levels of insurance resources and eligibility from state to state.


Comparative transcriptomic responses to chronic cadmium, fluoranthene, and atrazine exposure in Lumbricus rubellus.
Transcriptional responses of a soil-dwelling organism (the earthworm Lumbricus rubellus) to three chemicals, cadmium (Cd), fluoranthene (FA), and atrazine (AZ), were measured following chronic exposure, with the aim of identifying the nature of any shared transcriptional response. Principal component analysis indicated full or partial separation of control and exposed samples for each compound but not for the composite set of all control and exposed samples. Partial least-squares discriminant analysis allowed separation of the control and exposed samples for each chemical and also for the composite data set, suggesting a common transcriptional response to exposure. Genes identified as changing in expression level (by the least stringent test for significance) following exposure to two chemicals indicated a substantial number of common genes (> 127). The three compound overlapping gene set, however, comprised only 25 genes. We suggest that the low commonality in transcriptional response may be linked to the chronic concentrations (approximately 10% EC50) and chronic duration (28 days) used. Annotations of the three compound overlapping gene set indicated that genes from pathways most often associated with responses to environmental stress, such as heat shock, phase I and II metabolism, antioxidant defense, and cation balance, were not represented. The strongest annotation signature was for genes important in mitochondrial function and energy metabolism.


Sex pheromone of a coccoid insect with sexual and asexual lineages: fate of an ancestrally essential sexual signal in parthenogenetic females.
Sex pheromones play a central role in intersexual communication for reproduction in many organisms. Particularly in insects, reproductive isolation that leads to speciation is often achieved by shifts of pheromone chemistries. However, the divergence and evolution of pheromones remain largely unknown. This study reveals a unique evolutionary consequence for terpenoid pheromones in coccoid insects. Coccoids, such as mealybugs, show clear sexual dimorphism: males are dwarf and short-lived, whereas females are wingless and almost immobile. Female pheromones are therefore indispensable for males to navigate for sexual reproduction, but some females can reproduce asexually. Interestingly, a derived asexual lineage that reproduces by parthenogenesis coexists with its ancestral lineage that reproduces sexually in a population of the pineapple mealybug, Dysmicoccus brevipes Here, we isolated, characterized and synthesized a novel monoterpene, (-)-(anti-1,2-dimethyl-3-methylenecyclopentyl)acetaldehyde, as a pheromone of the sexual females of Dbrevipes This monoterpene aldehyde, with an irregular linkage of isoprene units, is notable, because all mealybug pheromones previously reported are carboxylic esters of terpenols. This compound was, however, never produced by the asexual females. As a consequence of acquiring parthenogenetic reproduction, the asexual females appear to have abandoned the production of the sex pheromone, which had been essential to attracting males in their ancestors.


Evidence-based alternative medicine?
The validity of evidence-based medicine (EBM) is the subject of ongoing controversy. The EBM movement has proposed a "hierarchy of evidence," according to which randomized controlled trials (RCTs) and meta-analyses of RCTs provide the most reliable evidence concerning the efficacy of medical interventions. The evaluation of alternative medicine therapies highlights problems with the EBM hierarchy. Alternative medical researchers-like those in mainstream medicine-wish to evaluate their therapies using methods that are rigorous and that are consistent with their philosophies of medicine and healing. These investigators have three ways to relate their work to EBM. They can accept the EBM hierarchy and carry out RCTs when possible; they can accept the EBM standards but argue that the special characteristics of alternative medicine warrant the acceptance of "lower" forms of evidence; or they can challenge the EBM approach and work to develop new research designs and new standards of evidence that reflect their approach to medical care. For several reasons, this last option is preferable. First, it will best meet the needs of alternative medicine practitioners. Moreover, because similar problems beset the evaluation of mainstream medical therapies, reevaluation of standards of evidence will benefit everyone in the medical community--including, most importantly, patients.


Elevated concentrations of nerve growth factor in heart and mesenteric arteries of spontaneously hypertensive rats.
Considerable evidence indicates an enhanced sympathetic innervation of muscular resistance arteries in the spontaneously hypertensive rat (SHR) compared with its normotensive Wistar-Kyoto (WKY) control. Since nerve growth factor (NGF) is known to affect the growth of sympathetic nerves, we have utilized a sensitive two-site enzyme linked immunoassay for NGF to compare the NGF content of hearts and mesenteric arteries of developing SHR and WKY rats. NGF levels in hearts revealed similar, although not identical, patterns of expression. In both strains, NGF levels declined from postnatal day 15, the earliest age examined, to stabilize at adult levels by postnatal day 32. Adult SHR concentrations were similar to those in age matched WKY controls. In contrast, in the mesenteric vascular bed, NGF levels of SHR were greater than those of WKY controls at all ages above 15 days. Moreover, these changes in NGF occurred concomitantly with increases in vascular mass and medial smooth muscle hyperplasia in the SHR. Whether abnormal NGF levels are a cause or consequence of vascular smooth muscle growth has yet to be determined. These results are consistent with the hypothesis that the hyper-noradrenergic innervation of SHR vascular tissues results from an early elevation of NGF gene expression.


Precision medicine in diabetes: an opportunity for clinical translation.
Metabolic disorders present a public health challenge of staggering proportions. In diabetes, there is an urgent need to better understand disease heterogeneity, clinical trajectories, and related comorbidities. A pressing and timely question is whether we are ready for precision medicine in diabetes. Some biological insights that have emerged during the last decade have already been used to direct clinical decision making, especially in monogenic forms of diabetes. However, much work is necessary to integrate high-dimensional explorations into complex disease architectures, less penetrant biological alterations, and broader phenotypes, such as type 2 diabetes. In addition, for precision medicine to take hold in diabetes, reproducibility, interpretability, and actionability remain key guiding objectives. In this review, we examine how mounting data sets generated during the last decade to understand biological variability are now inspiring new venues to clarify diabetes nosology and ultimately translate findings into more effective prevention and treatment strategies.


Characterization of a Propionibacterium acnes Surface Protein as a Fibrinogen-Binding Protein.
Propionibacterium acnes (P. acnes) is a major skin-associated bacterium that was long considered commensal, until several studies revealed it to be an opportunistic pathogen. We investigated the ability of P. acnes surface proteins to recognize ECM proteins and showed that a 58 kDa P. acnes surface protein was specifically recognized by human fibrinogen (hFg). The 58 kDa protein was further characterized by two-dimensional (2-D) electrophoresis and MALDI-ToF as a P. acnes host cell-surface attachment protein, PA25957, recognizing dermatan sulfate (DsA1). This protein sequence contains 432 amino acids with the presence of three structurally different domains: an N-terminal signal peptide, a C-terminal LPXTG motif, and a PT repeat region. DsA1 is mostly produced during stationary phase. It appears to be highly glycosylated, containing GalNAc residues. Purified DsA1 strongly recognizes the Aα and Bβ subunits of hFg, and specific enzymatic deglycosylation of hFg demonstrated the involvement of the protein backbone in the recognition process. The Bβ subunit of hFg was cloned in four peptide fractions (Fg1-Fg4). The N-terminal Fg1 peptide of hFg was recognized by DsA1, and priming DsA1 with Fg1 inhibited DsA1/hFg recognition. We describe here for the first time, the characterization of a P. acnes surface glycoprotein recognizing human fibrinogen.


2-Deoxy-d-glucose reduces epileptiform activity by presynaptic mechanisms.
2-Deoxy-d-glucose (2DG), a glucose analog that inhibits glycolysis, has acute and chronic antiepileptic effects. We evaluated 2DG's acute effects on synaptic and membrane properties of CA3 pyramidal neurons in vitro. 2DG (10 mM) had no effects on spontaneously occurring postsynaptic currents (PSCs) in 3.5 mM extracellular potassium concentration ([K+]o). In 7.5 mM [K+]o, 2DG significantly reduced the frequency of epileptiform bursting and the charge carried by postsynaptic currents (PSCs) with a greater effect on inward excitatory compared with outward inhibitory charge (71% vs. 40%). In 7.5 mM [K+]o and bicuculline, 2DG reduced significantly the excitatory charge by 67% and decreased the frequency but not amplitude of excitatory PSCs between bursts. In 7.5 mM [K+]o, 2DG reduced pharmacologically isolated inhibitory PSC frequency without a change in amplitude. The frequency but not amplitude of inward miniature PSCs was reduced when 2DG was applied in 7.5 mM [K+]o before bath application of TTX, but there was no effect when 2DG was applied after TTX, indicating a use-dependent uptake of 2DG was required for its actions at a presynaptic locus. 2DG did not alter membrane properties of CA3 neurons except for reducing the slow afterhyperpolarization in 3.5 but not 7.5 mM [K+]o. The reduction in frequency of spontaneous and inward miniature PSCs in elevated [K+]o indicates a presynaptic mechanism of action. 2DG effects required use-dependent uptake and suggest an important role for glycolysis in neuronal metabolism and energetics in states of high neural activity as occur during abnormal network synchronization and seizures. NEW & NOTEWORTHY 2-Deoxy-d-glucose (2DG) is a glycolytic inhibitor and suppresses epileptiform activity acutely and has chronic antiepileptic effects. The mechanisms of the acute effects are not well delineated. In this study, we show 2DG suppressed abnormal network epileptiform activity without effecting normal synaptic network activity or membrane properties. The effects appear to be use dependent and have a presynaptic locus of action. Inhibition of glycolysis is a novel presynaptic mechanism to limit abnormal neuronal network activity and seizures.


Continuous Suprascapular Nerve Block With a Perineural Catheter for Reverse Shoulder Arthroplasty Rescue Analgesia in a Patient With Severe Chronic Obstructive Pulmonary Disease.
Reverse open shoulder arthroplasty requires a comprehensive analgesic plan involving regional anesthesia. The commonly performed interscalene brachial plexus blockade confers a high likelihood of diaphragmatic paralysis via phrenic nerve palsy, making this option riskier in patients with limited pulmonary reserve. Continuous blockade of the suprascapular nerve, a more distal branch of the C5 and C6 nerve roots, may be a viable alternative. We report a successful case of the use of a suprascapular nerve block with continuous programmed intermittent bolus perineural analgesia in a patient with severe chronic obstructive pulmonary disease who underwent reverse open shoulder arthroplasty.


NZPrEP Demonstration Project: protocol for an open-label, single-arm trial of HIV pre-exposure prophylaxis (PrEP) to determine feasibility, acceptability, adverse and behavioural effects of PrEP provision to gay and bisexual men in publicly funded sexual health clinics in Auckland, New Zealand.
New Zealand has experienced a rise in HIV diagnoses in recent years and new interventions are required to address this. NZPrEP (A demonstration project of HIV preexposure prophylaxis in Aotearoa New Zealand) is an open-label, single-arm treatment evaluation study to investigate feasibility, retention, adherence, and clinical and behavioural outcomes of HIV pre-exposure prophylaxis (PrEP) provision to gay and bisexual men (GBM) in a publicly funded secondary sexual health service in Auckland, New Zealand. The sample size is 150 GBM. Inclusion criteria were specific behavioural risk factors indicating an increased risk of HIV infection. Exclusion criteria were hepatitis B infection, any medical contraindications to prescribing tenofovir/emtricitabine or factors limiting ability to adhere to the study protocol. Eligible participants will be screened for HIV and other sexually transmissible infections (STIs) and for any medical contraindications to PrEP, and enrolled for a maximum follow-up period of 96 weeks. They will be required to attend for 3-monthly testing for HIV and STIs and monitoring for renal and liver toxicity. Participants will also be required to complete an online behavioural survey after each study visit. The outcomes of interest are feasibility of PrEP provision in a sexual health clinic setting, PrEP acceptability, and adverse medical and behavioural effects of PrEP. The study sample is limited to 150 participants due to funding and service constraints. Statistical analysis of all primary and secondary outcomes will be performed using Stata V.14 at the University of Auckland. Results for primary and secondary endpoints will be reported after the conclusion of the study in March 2019. The study was approved by the Health and Disability Ethics Committee on 15 September 2016 (16/NTA/112). Key findings will be submitted to peer-reviewed journals. A summary report will be circulated to the study and community stakeholders, and to the Auckland District Health Board, Ministry of Health and Pharmac. ACTRN12616001387415; Pre-results.


Integrative Treatment of Personality Disorder. Part I: Psychotherapy.
In this paper, we outline the concept of integrative therapy of borderline personality, also referred to as fragmented personality, which we consider to be the core psychopathology underlying all clinical subtypes of personality disorder. Hence, the terms borderline personality, borderline disorder, fragmented personality, and personality disorder are used interchangeably, as synonyms. Our integrative approach combines pharmacotherapy and psychotherapy, each specifically tailored to accomplish a positive feedback modulation of their respective effects. We argue that pharmacotherapy and psychotherapy of personality disorder complement each other. Pharmacological control of disruptive affects clears the stage, in some cases builds the stage, for the psychotherapeutic process to take place. In turn, psychotherapy promotes integration of personality fragments into more cohesive structures of self and identity, ultimately establishing self-regulation of mood and anxiety. We introduce our original method of psychotherapy, called reconstructive interpersonal therapy (RIT). The RIT integrates humanistic-existential and psychodynamic paradigms, and is thereby designed to accomplish a deep reconstruction of core psychopathology within the setting of high structure. We review and comment the current literature on the strategies, goals, therapy process, priorities, and phases of psychotherapy of borderline disorders, and describe in detail the fundamental principles of RIT.


Cell cycle arrest in Metformin treated breast cancer cells involves activation of AMPK, downregulation of cyclin D1, and requires p27Kip1 or p21Cip1.
The antihyperglycemic drug metformin may have beneficial effects on the prevention and treatment of cancer. Metformin is known to activate AMP-activated protein kinase (AMPK). It has also been shown to inhibit cyclin D1 expression and proliferation of some cultured cancer cells. However, the mechanisms of action by which metformin mediates cell cycle arrest are not completely understood. In this study, metformin was found to inhibit proliferation of most cultured breast cancer cell lines. This was independent of estrogen receptor, HER2, or p53 status. Inhibition of cell proliferation was associated with arrest within G0/G1 phase of the cell cycle. As in previous studies, metformin treatment led to activation of (AMPK) and downregulation of cyclin D1. However, these events were not sufficient for cell cycle arrest because they were also observed in the MDA-MB-231 cell line, which is not sensitive to growth arrest by metformin. In sensitive breast cancer lines, the reduction in cyclin D1 led to release of sequestered CDK inhibitors, p27Kip1 and p21Cip1, and association of these inhibitors with cyclin E/CDK2 complexes. The metformin-resistant cell line MDA-MB-231 expresses significantly lower levels of p27Kip1 and p21Cip1 than the metformin-sensitive cell line, MCF7. When p27Kip1 or p21Cip1 were overexpressed in MDA-MB-231, the cells became sensitive to cell cycle arrest in response to metformin. Cell cycle arrest in response to metformin requires CDK inhibitors in addition to AMPK activation and cyclin D1 downregulation. This is of interest because many cancers are associated with loss or downregulation of CDK inhibitors and the results may be relevant to the development of anti-tumor reagents that target the AMPK pathway.


C=C π Bond Modified Graphitic Carbon Nitride Films for Enhanced Photoelectrochemical Cell Performance.
Applications of graphitic carbon nitride (g-CN) in photoelectrochemical and optoelectronic devices are still hindered due to the difficulties in synthesis of g-CN films with tunable chemical, physical and catalytic properties. Herein we present a general method to alter the electronic and photoelectrochemical properties of g-CN films by annealing. We found that N atoms can be removed from the g-CN networks after annealing treatment. Assisted by theoretical calculations, we confirm that upon appropriate N removal, the adjacent C atoms will form new C=C π bonds. Detailed calculations demonstrate that the highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) are located at the structure unit with C=C π bonds and the electrons are more delocalized. Valence band X-ray photoelectron spectroscopy spectra together with optical absorption spectra unveil that the structure changes result in the alteration of the g-CN energy levels and position of band edges. Our results show that the photocurrent density of the annealed g-CN film is doubled compared with the pristine one, thanks to the better charge separation and transport within the film induced by the new C=C π bonds. An ultrathin TiO2 film (2.2 nm) is further deposited on the g-CN film as stabilizer and the photocurrent density is kept at 0.05 mA cm-2 at 1.23 V versus reversible hydrogen electrode after two-cycle stability assessment. This work enables the applications of g-CN films in many electronic and optoelectronic devices.


Intricate Conformational Tunneling in Carbonic Acid Monomethyl Ester.
Disentangling internal and external effects is a key requirement for understanding conformational tunneling processes. Here we report the s- trans/ s- cis tunneling rotamerization of carbonic acid monomethyl ester (1) under matrix isolation conditions and make comparisons to its parent carbonic acid (3). The observed tunneling rate of 1 is temperature-independent in the 3-20 K range and accelerates when using argon instead of neon as the matrix material. The methyl group increases the effective half life (τeff) of the energetically disfavored s- trans-conformer from 3-5 h for 3 to 11-13 h for 1. Methyl group deuteration slows the rotamerization further (τeff ≈ 35 h). CCSD(T)/cc-pVQZ//MP2/aug-cc-pVTZ computations of the tunneling probability suggest that the rate should be almost unaffected by methyl substitution or its deuteration. Thus the observed relative rates are puzzling, and they disagree with previous explanations involving fast vibrational relaxation after the tunneling event facilitated by the alkyl rotor.


Independent variation in the number of coated pits and of coated vesicles in cultured fibroblasts.
When tissue culture cells were maintained at 37 degrees C in a serum-free medium for 4 hr no change in the number of coated pits could be detected using ultrastructural techniques. However, the number of coated vesicles was highly significantly increased, being 179% more than in the control cultures. If the cells were put back into a medium supplemented with 5% calf serum, the number of coated pits was unchanged, but the number of coated vesicles decreased and returned to the control level within a few minutes. The same results were obtained when using ligands such as Low Density Lipoprotein or alpha-2-macroglobulin which are known to be internalized via coated structures. It is concluded that coated pits appear and disappear at equal rates and that coated vesicles can accumulate independently. It is suggested that this could be due to the presence of a large reserve of soluble clathrin. This pool would have a low turnover rate because cycloheximide did not block coated vesicle accumulation over the period studied.


[Flow rates in the isolated perfused rat liver by pulse labeling with radioactive substrates and mathematical analysis of the wash-out kinetics].
The multiple indicator dilution technique (Goresky and Bach) is critically evaluated and its application to the isolated, hemoglobin-free perfused rat liver is described. From the results of pulse labelling experiments using indicator substances for the total aqueous space of the liver ([3H]water and [14C]urea) an for the extracellular space ([14C]sucrose, [3H]-inulin and [3H]dextrane), it is concluded that the mathematical model of the liver in situ according to Goresky and Bach is sufficient to also describe the hemodynamics of the isolated liver perfused with a saline solution. The data indicate that the multiple indicator dilution technique in combination with the available mathematical basis is applicable to the study of transport across the liver cell membrane. The method, however, is restricted to compounds which are metabolized very slowly, such as D-lactate. A possible extension of this method to metabolic processes is discussed in view of pulse labelling experiments with [3H]-and [1-14C]L-lactate. Since the involvement of metabolism in the available mathematical model is not differentiated and the release of products is not taken into consideration, the method in its present state is not applicable to studies of metabolism. Moreover, even the parameters of transport derived by this technique are of limited value, when the transported compounds are rapidly metabolized by reversible reactions. Despite these uncertainties, the present data indicate that the transport of L-lactate is ten times faster than that of D-lactate.


Introductory campaign.
When University of Maryland Medical Center was about to open its new orthopaedic acute-care unit, it used the news to recruit folks to join the staff.


Detection of air trapping on inspiratory and expiratory phase images obtained by 0.3-second cine CT in the lungs of free-breathing young children.
The objective of our study was to evaluate whether 0.3-second cine CT can be used to detect air trapping in the lungs of young children. In 30 children (mean age, 25 months), 0.3-second cine CT was performed at six levels during 3 seconds of quiet breathing. The study population was divided into an air trapping group (n = 24) and a no-air trapping group (n = 6). Lung density was measured at an abnormal area (with or without air trapping) and an adjacent normal area on inspiratory and expiratory phase images. Lung density differences between inspiration and expiration were calculated and compared in abnormal areas (with or without air trapping) and in normal areas. Their percentages were calculated and compared between the two groups. In addition, lung density differences between abnormal and adjacent normal areas were calculated and compared between the two groups. Lung density differences between inspiration and expiration were smaller in areas with air trapping (mean +/- SD, -19 +/- 34 H) than in abnormal areas without air trapping (138 +/- 36 H) (p < 0.001) or in normal areas (111 +/- 49 H) (p < 0.001). Their percentages were smaller in the group with air trapping (-27% +/- 54%) than in the group with no air trapping (120% +/- 87%) (p < 0.001). In the group with air trapping, lung density differences were larger at the expiratory phase (260 +/- 77 H) than at the inspiratory phase (129 +/- 69 H) (p < 0.001), but did not change through the respiratory cycle in the group with no air trapping (p > 0.05). Air trapping can be accurately detected in the lungs of free-breathing young children using 0.3-second cine CT.


[Perforation of a duodenal stress ulcer in a 7-year-old polytraumatized boy].
This is a report of a seven-year-old polytraumatised boy with perforation of a duodenal stress-ulcer. Aspects of the etiology and pathogenesis were discussed. The main therapeutical efforts by stress-ulcer risk patients consist in an early sufficient shock therapy and adequate stress-ulcer prophylaxis to prevent the origin of stress-ulcers and their complications like bleeding and perforation.


[Significance of p53 protein expression and p53 gene mutation in so-called pulmonary sclerosing hemangioma].
So-called pulmonary sclerosing hemangioma (PSH) is an uncommon tumor of the lung and its histogenesis and origin are uncertain to date. A general consensus appears to have been reached that PSH is a benign neoplasm, but a few PSHs are found to invade and metastasize. The expression of p53 protein and mutation of p53 gene are significant parameters which can reflect biological behavior of tumor cells. The aim of this study is to investigate the p53 protein expression and p53 gene mutation in PSH, and explore their significance. The expression of p53 protein and mutation of p53 gene were examined in polygonal cells and cuboidal cells of PSH by immunohistochemical method, laser capture microdissection (LCM), single-stranded conformation polymorphism (SSCP) and DNA sequencing. The positive rate of p53 protein was 21.1% (4/19) and the mutation rate of p53 gene was 26.3% (5/19) by SSCP and 42.1% (8/19) by DNA sequencing respectively. In 4 cases of immunopositive PSH tissues, 2 were missense mutations, and 1 was both missense and frameshift mutation. Out of 15 cases of immunonegative PSH tissues, 4 were frameshift mutations and 1 was missense mutation. Of 8 PSH tissues with p53 gene mutation, 5 were identified in only polygonal cells, and 2 in only cuboidal cells and 1 in both polygonal and cuboidal cells. The expression of p53 protein may not be indicative of p53 gene mutation in PSH. The alteration of p53 gene and the expression of p53 protein are identified in both polygonal and cuboidal cells. The high mutation rate of p53 gene may indicate that PSH has potentially malignant biological behavior.


Optimization of the porous structure and polarity of polymethacrylate-based monolithic capillary columns for the LC-MS separation of enzymatic digests.
The porous structure as well as the polarity of methacrylate ester-based monolithic stationary phases has been optimized to achieve the separation of various peptides originating from enzymatic digestion. The porous structure, determined by the size of both pores and microglobules, was varied through changes in the composition of porogenic solvents in the polymerization mixture, while the polarity was controlled through the incorporation of butyl, lauryl, or octadecyl methacrylate in the polymer backbone. Both the morphology and the chemistry of the monoliths had a significant effect on the retention and efficiency of the capillary columns. The best resolution of peptidic fragments obtained by digestion of Cytochrome c with trypsin in solution was obtained in a gradient LC-MS mode using a monolithic capillary column of poly(lauryl methacrylate-co-ethylene dimethacrylate) featuring small pores and small microglobules. Raising the temperature from 25 to 60 degrees C enabled separations to be carried out at 40% higher flow rates. Separations carried out at 60 degrees C with a steeper gradient proceeded without loss of performance in half the time required for a comparable separation at room temperature. Our preparation technique affords monolithic columns with excellent column-to-column and run-to-run repeatability of retention times and pressure drops.


Differential binding and activation of thyroid hormone response elements by TR alpha 1 and RXR alpha-trap heterodimers.
Thyroid hormone receptor (TR) forms homo- and heterodimers on various thyroid hormone response elements (TREs). We wished to clarify the relationship of homo- and heterodimer binding to TREs and their trans-activation. We investigated binding characteristics in gel mobility shift assays using synthetic direct repeat (DR) TREs having the consensus motifs separated by different oligonucleotide gaps, and we compared binding to trans-activation mediated via the direct repeat TRE. HTR alpha 1 purified from E. coli formed a monomer and homodimer on DR-TRE +0 to +5 but binding did not closely correlate with T3-dependent trans-activation. When RXR alpha expressed in COS 1 cell was added to purified TR alpha 1 in the gel shift assays, TR/RXR heterodimers were formed, and binding of heterodimers correlated highly with the level of trans-activation. These results strongly suggest that TR/TRAP heterodimers mediate the effect of thyroid hormone on DR-TREs. We also found T3-dependent disruption of homodimer formation on DR +0 to +2 and that T3 increased heterodimer formation on these TREs.


Assessment of CASP10 contact-assisted predictions.
In CASP10, for the first time, contact-assisted structure predictions have been assessed. Sets of pairs of contacting residues from target structures were provided to predictors for a second round of prediction after the initial round in which they were given only sequences. The objective of the experiment was to measure model quality improvement resulting from the added contact information and thereby assess and help develop so-called hybrid prediction methods--methods where some experimentally determined distance constraints are used to augment de novo computational prediction methods. The results of the experiment were, overall, quite promising.


Angiotensin-converting activity of tissue kallikrein.
This study examined the ability of tissue kallikreins, purified from both rat submandibular gland (SMG) and human urine, to form angiotensin II from synthetic angiotensin I. Both kallikreins converted angiotensin I to angiotensin II at neutral pH with the following kinetic constants: SMG kallikrein, Km = 9.43 x 10(-5) mol/l, Kcat = 1.58 mumols/mg protein/min; human urinary kallikrein, Km = 1.71 x 10(-4) mol/l, Kcat = 0.06 mumol/mg protein/min. These activities were not affected by angiotensin-converting enzyme (ACE) inhibitor. These results suggest that tissue kallikrein might participate in the formation of angiotensin II during administration of an ACE inhibitor.


M1/M2 macrophage polarity in normal and complicated pregnancy.
Tissue macrophages play an important role in all stages of pregnancy, including uterine stromal remodeling (decidualization) before embryo implantation, parturition, and post-partum uterine involution. The activation state and function of utero-placental macrophages are largely dependent on the local tissue microenvironment. Thus, macrophages are involved in a variety of activities such as regulation of immune cell activities, placental cell invasion, angiogenesis, and tissue remodeling. Disruption of the uterine microenvironment, particularly during the early stages of pregnancy (decidualization, implantation, and placentation) can have profound effects on macrophage activity and subsequently impact pregnancy outcome. In this review, we will provide an overview of the temporal and spatial regulation of utero-placental macrophage activation during normal pregnancy in human beings and rodents with a focus on more recent findings. We will also discuss the role of M1/M2 dysregulation within the intrauterine environment during adverse pregnancy outcomes.


The first description of a hormone-sensitive lipase from a basidiomycete: Structural insights and biochemical characterization revealed Bjerkandera adusta BaEstB as a novel esterase.
The heterologous expression and characterization of a Hormone-Sensitive Lipases (HSL) esterase (BaEstB) from the Basidiomycete fungus Bjerkandera adusta is reported for the first time. According to structural analysis, amino acid similarities and conservation of particular motifs, it was established that this enzyme belongs to the (HSL) family. The cDNA sequence consisted of 969 nucleotides, while the gene comprised 1133, including three introns of 57, 50, and 57 nucleotides. Through three-dimensional modeling and phylogenetic analysis, we conclude that BaEstB is an ortholog of the previously described RmEstB-HSL from the phylogenetically distant fungus Rhizomucor miehei. The purified BaEstB was characterized in terms of its specificity for the hydrolysis of different acyl substrates confirming its low lipolytic activity and a noticeable esterase activity. The biochemical characterization of BaEstB, the DLS analysis and the kinetic parameters determination revealed this enzyme as a true esterase, preferentially found in a dimeric state, displaying activity under alkaline conditions and relative low temperature (pH = 10, 20°C). Our data suggest that BaEstB is more active on substrates with short acyl chains and bulky aromatic moieties. Phylogenetic data allow us to suggest that a number of fungal hypothetical proteins could belong to the HSL family.


The Spin-Flip Variant of the Algebraic-Diagrammatic Construction Yields the Correct Topology of S1/S0 Conical Intersections.
While the conventional variants of the algebraic-diagrammatic construction (ADC) scheme for the polarization propagator are generally incapable of correctly describing the topology of S1/S0 conical intersections (CIs), its corresponding spin-flip (SF) variant of third-order ADC (ADC(3)) is herein demonstrated to successfully reproduce the S1/S0 minimum-energy CI (MECI) of twisted formaldinium (H2C═NH2+). Analytical nuclear excited-state gradients of ADC have been used in combination with the CIOpt program for the optimization of the MECI without the need for nonadiabatic-coupling vectors. For comparison, MS-CASPT2 calculations were performed via conventional CI optimization employing analytical nonadiabatic-coupling vectors. It is shown that SF-ADC(3) yields the correct dimensionality of the CI and overall compares very favorably to the MS-CASPT2 results.


Clinical Instructors' Perspectives on the Assessment of Clinical Knowledge of Undergraduate Nursing Students: A Descriptive Phenomenological Approach.
This study aimed to determine clinical instructors' perceptions of the assessments used to evaluate the clinical knowledge of undergraduate nursing students. This study uses a descriptive phenomenological approach. Purposive sampling was used to recruit sixteen clinical instructors for semi-structured interviews between August and December 2019. All interviews were audio recorded and transcribed verbatim. Data were analyzed using a modified Colaizzi's seven-step method. Four criteria were used to ensure the study's validity: credibility, transferability, dependability, and confirmability. Three themes were identified in the clinical instructors' views on evaluating the clinical performance of student nurses: familiarity with students, patchwork clinical learning, and differing perceptions of the same scoring system. The study results suggest a need for a reliable, valid, and consistent approach to evaluating students' clinical knowledge. If the use of patchwork clinical internships for student nurses is unavoidable, a method for assessing student nurses' clinical performance that requires instructor consensus is necessary.


Self-assembly of coiled-coil tetramers in the 1.40 A structure of a leucine-zipper mutant.
The hydrophobic core of the GCN4 leucine-zipper dimerization domain is formed by a parallel helical association between nonpolar side chains at the a and d positions of the heptad repeat. Here we report a self-assembling coiled-coil array formed by the GCN4-pAe peptide that differs from the wild-type GCN4 leucine zipper by alanine substitutions at three charged e positions. GCN4-pAe is incompletely folded in normal solution conditions yet self-assembles into an antiparallel tetraplex in crystals by formation of unanticipated hydrophobic seams linking the last two heptads of two parallel double-stranded coiled coils. The GCN4-pAe tetramers in the lattice associate laterally through the identical interactions to those in the intramolecular dimer-dimer interface. The van der Waals packing interaction in the solid state controls extended supramolecular assembly of the protein, providing an unusual atomic scale view of a mesostructure.


[Problems and solutions in the use of RAPD to the identification of the Chinese drugs "xi-xin" (Herba asari) and its substitutes].
The use of random amplified polymorphic DNAs (RAPD) on the identification of crude drugs has a few special problems, e.g., the influence of the degree of degradation of DNA templates on the amplified products, the reliability of results by one or two primers and the application range of RAPD, etc., so the problems were discussed through researches on Herba Asari. In this experiment, the DNA of twenty-one samples from different locations and collected at different time were extracted. The suitable concentrations of DNA templates and the suitable primers were selected. After this, the DNA of these samples were amplified by RAPD. The results showed that the concentration and the degree of degradation of DNA templates, and the different sources of crude drugs affected the result of RAPD assay. Some suggestions were made to solve these problems, such as the selection of DNA templates concentration, the screening of suitable primers, and the use of contrast groups and cluster analysis. Furthermore, the application range of RAPD to the identification of crude drugs was discussed. All of these will provide the guide to the use of RAPD for the identification of crude drugs.


Simultaneous targeting of CD14 and factor XIa by a fusion protein consisting of an anti-CD14 antibody and the modified second domain of bikunin improves survival in rabbit sepsis models.
Severe sepsis is a complex, multifactorial, and rapidly progressing disease characterized by excessive inflammation and coagulation following bacterial infection. To simultaneously suppress pro-inflammatory and pro-coagulant responses, we genetically engineered a novel fusion protein (MR1007) consisting of an anti-CD14 antibody and the modified second domain of bikunin, and evaluated the potential of MR1007 as an anti-sepsis agent. Suppressive effects of MR1007 on lipopolysaccharide (LPS)-induced inflammatory responses were assessed using peripheral blood mononuclear cells or endothelial cells. Its inhibitory activity against the coagulation factor XIa was assessed using a purified enzyme and a chromogenic substrate. Anticoagulant activity was assessed using human or rabbit plasma. Anti-inflammatory and anti-coagulant effects and/or survival benefits were evaluated in an endotoxemia model and a cecal ligation and puncture model. MR1007 inhibited LPS-induced cytokine production in peripheral blood mononuclear cells and endothelial cells, inhibited factor XIa, and exhibited anticoagulant activity. In an endotoxemia model, 0.3-3mg/kg MR1007 suppressed pro-inflammatory and pro-coagulant responses in a dose-dependent manner; at a dose of 3mg/kg, the protein improved survival even when administered 8h after the LPS injection. In addition, 10mg/kg MR1007 administered 2h post cecal ligation and puncture improved survival. However, MR1007 administered at doses up to 30mg/kg did not increase ear bleeding time or bacterial counts in the cecal ligation and puncture model. Thus, simultaneous targeting of CD14 and factor XIa improves survival in the rabbit endotoxemia and sepsis models and represents a promising approach for the treatment of severe sepsis.


Finite-size analysis in neural network classification of critical phenomena.
We analyze the problem of supervised learning of ferromagnetic phase transitions from the statistical physics perspective. We consider two systems in two universality classes, the two-dimensional Ising model and two-dimensional Baxter-Wu model, and perform careful finite-size analysis of the results of the supervised learning of the phases of each model. We find that the variance of the neural network (NN) output function (VOF) as a function of temperature has a peak in the critical region. Qualitatively, the VOF is related to the classification rate of the NN. We find that the width of the VOF peak displays the finite-size scaling governed by the correlation length exponent ν of the universality class of the model. We check this conclusion using several NN architectures-a fully connected NN, a convolutional NN, and several members of the ResNet family-and discuss the accuracy of the extracted critical exponents ν.


Dichlorobis(2-phenylazopyridine)ruthenium(II) complexes: characterisation, spectroscopic and structural properties of four isomers.
The didentate ligand 2-phenylazopyridine (azpy) can--in theory--give rise to five different isomeric complexes of the type [Ru(azpy)2Cl2], of which three have been known since 1980. The molecular structures of the cis-dichlorobis(2-phenylazopyridine) ruthenium(II) complexes alpha-[Ru(azpy)2Cl2] and beta-[Ru(azpy)2Cl2](in which the coordinating pyridine nitrogen atoms are in mutually trans and cis positions, respectively, whilst the azo nitrogen atoms are in mutually cis positions) were unambiguously determined in the early 1980s. The third isomer, gamma-[Ru(azpy)2Cl2], has for two decades, erroneously, been assumed to be the all-trans isomer. In a recent communication we have proven that for this gamma isomer the chloride ions are indeed in a trans geometry, but the pyridine nitrogen and azo nitrogen atoms of the two azpy ligands are in mutually cis geometries. In this paper the isolation of a fourth isomer is presented, the hitherto unknown delta-[Ru(azpy)2Cl2]. The isomeric structure of delta-[Ru(azpy)2Cl2] has been determined by 1H-NMR spectroscopy and single-crystal X-ray diffraction analysis, and is the all-trans isomer. The bis(azpy)-ruthenium(II) isomers are of interest because of the pronounced cytotoxicity they exhibit against tumour cell lines and could be very useful in the search for structure-activity relationships of antitumour-active ruthenium complexes, as among the isomers there is a significant difference in activity. It is of paramount importance to have a good understanding of the structural and spectroscopic properties of these complexes, which in this paper are compared and discussed, with a particular emphasis on 1D and 2D 1H NMR spectroscopies.


Improved survival of uncemented versus cemented femoral stems in patients aged < 70 years in a community total joint registry.
Aseptic loosening of the femoral stem remains a significant reason for revision in total hip arthroplasty (THA). Although stem fixation methods have changed over time, there is relatively little evidence supporting cemented or uncemented stems as more durable constructs. We examined whether there was a difference in survival to revision between cemented and uncemented THA stems (1) for any reason; (2) for aseptic loosening or loosening related to wear/osteolysis; (3) based on patient age groupings (as a proxy for patient activity level); and (4) based on procedural timeframe groupings between cemented and uncemented stems. A total of 6498 primary cemented and uncemented THAs were registered in our community total joint replacement registry between 1991 and 2011. Analysis was performed to compare age, sex, procedural timeframe, and diagnosis for both groups. Our primary outcome was revision of the stem component for aseptic loosening or loosening secondary to wear/osteolysis. Analyses were done using Wilcoxon rank sum tests, Pearson's chi-square tests, Kaplan Meier methods, and Cox regression. After adjusting for age, sex, primary diagnosis, and procedural timeframe as confounders, cemented femoral stem components were 1.63 times as likely as uncemented stems to be revised for any reason (p = 0.02) and 3.76 times as likely as uncemented stems to be revised for aseptic loosening or loosening related to wear/osteolysis (p < 0.001). When grouped by age, specifically in regard to revisions for aseptic loosening or loosening related to wear/osteolysis, uncemented stems had lower cumulative revision rates in patients aged < 70 years (p < 0.001) compared with cemented stems. There was a trend away from cemented fixation in our registry, which shifted from over 80% cemented stem use in 1996 to 3% in 2011. We found that uncemented stems were associated with fewer revisions for aseptic loosening in patients < 70 years old, but when all reasons for revision were considered, neither group demonstrated superior survival. With a mean followup of 6.5 years, longer followup is needed to verify these results over time.


Instabilities and pattern transformation in periodic, porous elastoplastic solid coatings.
Pattern transformation in periodic microporous elastoplastic solid coatings is caused by a buckling of the struts and a rotation of the nodes under compressive stresses. The results of a nonlinear numerical investigation confirm the critical role of the bifurcation of the periodic solid under compressive stresses. In striking contrast to the earlier observations of elastic instabilities in porous elastomeric solids, the elastic-plastic nature of the cross-linked periodic microstructure studied here provides the ability to lock in the transformed pattern with complete relaxation of the internal stresses. The study unveils a novel deformation mode in porous periodic solids in the form of organized buckling instability of weak strut elements.


Complex 3D bioprinting methods.
3D bioprinting technology is evolving in complexity to enable human-scale, high-resolution, and multi-cellular constructs to better mimic the native tissue microenvironment. The ultimate goal is to achieve necessary complexity in the bioprinting process to biomanufacture fully-functional tissues and organs to address organ shortage and lack of patient-specific disease models. In this Review, we presented an in-depth overview of complex 3D bioprinting approaches including evolution of complex bioprinting, from simple gel-casting approach to multi-material bioprinting to omnidirectional bioprinting approaches, and emerging bioprinting approaches, including 4D bioprinting and in situ bioprinting technologies.


[E(μ-NBbp)]2 (E = P, As) - group 15 biradicals synthesized from acyclic precursors.
The synthesis of Bbp stabilized biradicals of the type [E(μ-NBbp)]2 (E = P, As) (Bbp = 2,6-bis[bis(trimethylsilyl)methyl]phenyl) was investigated. Contrary to the established synthetic protocol for terphenyl substituted biradicals [E(μ-NTer)]2 by reduction of [ClE(μ-NTer)]2, the analogous Bbp substituted precursor [ClE(μ-NBbp)]2 could only be obtained in low yields. For this reason, a new synthesis had to be found to generate [E(μ-NBbp)]2. Instead of using cyclic [ClE(μ-NBbp)]2 we prepared the acyclic Bbp-N(ECl2)2 species starting from Bbp-NH2 and ECl3 and studied the reduction with magnesium chips, which led to the formation of the desired biradicals [E(μ-NBbp)]2. All species along the reaction path were fully characterized.


High-level recombinant protein production in bioreactors using the baculovirus-insect cell expression system.
In order to develop an efficient process for large-scale production of recombinant protein, various factors were studied which affect the productivity of Sf-9 (Spodoptera frugiperda) insect cells when using the baculovirus expression system. It was shown that upon infection with the Bac-BRV6L recombinant baculovirus, the level per cell of VP6 (a bovine rotavirus nucleocapsid protein) would drop 10-fold when host cell density at the time of infection increased from 2 x 10(6) to 3 x 10(6) cells/mL. The decrease was found to be totally reversible by culture medium renewal after infection, even when cells were infected at the stationary phase. Recombinant protein production was 4-6 times higher using TNMFH medium supplemented with 10% fetal bovine serum (FBS) than in IPL/41 serum-free medium. Fine-tuning of infection parameters in a 4-L surface-aerated bioreactor resulted in the production of typically 350 mg/L of VP6 protein, representing more than 25% of total cell proteins.


Gas6 Attenuates Sepsis-Induced Tight Junction Injury and Vascular Endothelial Hyperpermeability via the Axl/NF-κB Signaling Pathway.
Vascular endothelial functional dysregulation and barrier disruption are involved the initiation and development of sepsis. Growth arrest-specific protein 6 (Gas6), one of the endogenous ligands of TAM receptors (Tyro3, Axl, and Mertk), is confirmed to have beneficial functions in hemostasis, inflammation, and cancer growth. Here, we demonstrated the protective effects of Gas6 on multi-organ dysfunction syndrome (MODS) in sepsis and the underlying mechanisms. We investigated Gas6-ameliorated MODS by inhibiting vascular endothelial hyperpermeability in a mouse model of sepsis. Additionally, in vitro, under lipopolysaccharide (LPS) stimulation in vascular endothelial cells, Gas6 attenuated vascular endothelial hyperpermeability by reinforcing the tight junction proteins occludin, zonula occludens-1 (ZO-1), and claudin5. Furthermore, Gas6 substantially suppressed NF-κB p65 activation. In addition, blocking the Gas6 receptor, Axl, partially reduced the protective effect of Gas6 on the vascular endothelial barrier and diminished the inhibitive effect of Gas6 on NF-κB p65 activation. Taken together, this study suggests that Gas6 has a protective effect on MODS in sepsis by inhibiting the vascular endothelial hyperpermeability and alteration of tight junction and that the Axl/NF-κB signaling pathway underlies these effects.


[Vaginal delivery in breech presentation does not increase early newborn morbidity. Results of 423 infants with breech presentation 1988-1992].
423 deliveries by breech presentation (1988-1992), delivered by vaginally or abdominally management, were analysed. 72 preterm (32nd to 36th gestational week) and 351 term labours were studied. In 239/423 (56.5%) cases neonates were delivered vaginally, in 54/423 (12.8%) and 130/423 (30.7%) cases primary or secondary cesarean section was necessary. In 120/423 (51.9%) cases of all primaparous spontaneously delivery were realized. There was no difference in early morbidity of vaginal und abdominal delivered neonates (pHNA, APGAR-Score, intracranially bleeding). Three intracranially bleedings (I degrees, II degrees, III degrees) observed in the vaginally delivery group (n = 239), were not the result of breech presentation or vaginally management. No correlation between vaginally management, acidosis and intracranially bleeding were observed. In patients presented with preterm labour (32nd to 36th gestational week) and/or fetal intrauterin growth retardation, cesarean section could be indicated. In postnatal sonographic screening of all neonates hip joint disorders were twice more frequently in female neonates (n = 51 vs. n = 26) without correlation to delivery mode. In conclusion, if certain personally and equipmently conditions are considered, vaginally delivery mode does not correlate to an increase of early neonatal morbidity.


Hepatic encephalopathy is related to anemia and fat-free mass depletion in liver transplant candidates with cirrhosis.
Although muscle wasting may lead to decreased ammonia detoxification in cirrhosis, the potential role of lean mass depletion in hepatic encephalopathy (HE) has not been explored. Anemia, hormonal abnormalities, and psychological distress may contribute to cognitive dysfunction, but data on their potential relation to HE are limited. Data on 108 cirrhotic liver transplant candidates enrolled in a prospective study on fatigue were retrospectively analyzed. HE was assessed clinically and with the number connection tests (NCT) A and B. Psychosocial distress was assessed with a validated questionnaire. Fasting serum glucose, insulin, ammonia, and the glomerular filtration rate (GFR) were measured. Fat and fat-free mass was evaluated with dual-energy X-ray absorptiometry. Serum cortisol, testosterone, dehydroepiandrosterone, thyroid function tests, interleukin-6, and tumor necrosis factor-α (TNF-α) were measured in a subgroup of 80 patients. A total of 28% of patients had (overt or minimal) HE. Anemia was present in 59%, diabetes in 29%, renal impairment in 16%, and fat-free mass depletion in 14%. In multivariate analysis, fat-free mass depletion was an independent predictor of HE and NCT-A; renal impairment of NCT-A and -B; and anemia of NCT-B (p < 0.05 for all). HE was also independently related to international normalized ratio and TNF-α (p < 0.05 for both), but not to other hormonal abnormalities or psychological distress. Plasma ammonia was independently associated to anemia (beta = 15.24, p = 0.049), fasting insulin (beta = 0.26, p < 0.05), and GFR (beta = -0.43, p = 0.003). Anemia and fat-free mass depletion are predictors of HE in cirrhotic liver transplant candidates along with liver failure, renal impairment, and systemic inflammation.


In vitro study of silica nanoparticle-induced cytotoxicity based on real-time cell electronic sensing system.
The biological fate and potential toxicity of nanoparticles have become hot topics since the use of nanoparticles was extended to the biomedical and biotechnological field. In this paper, an in vitro study of silica (SiO2) nanoparticle-induced cytotoxicity was presented mainly by real-time cell electronic sensing (RT-CES) system, which could be used for label-free, dynamic measurement of cell responses to cytotoxicants. The cytotoxicity of both 20-nm silica nanoparticles and microscale SiO2 powder in cultured RAW264.7 cells at dosage levels from 31.25 to 8000 microg/ml was evaluated comparatively. The experimental results showed that the cell index (CI) decrease induced by SiO2 nanoparticles appeared earlier than that by the microscale SiO2 powder at the same dose. The nanoparticles in the dose of 500 micdrog/ml had the toxic effect by the Cell Index Curves of RT-CES, while the microscale SiO2 powders in the same dose did not show obvious effect. The above experimental results were compared with the experiments of a tetrazolium compound-based colorimetric method (MTT assay) and LDH activity in medium. The results showed that the RT-CES assay displayed equal sensitivity and correlation to MTT assay. LDH activity caused by SiO2 nanoparticles at dose of 250 and 500 microg/ml was higher than that by microscale SiO2 particles, which indicated that the exposure to SiO2 nanoparticles could result in a dose-dependent cytotoxicity in cultured RAW264.7 cells due to cell membrane injury. The RT-CES system allows a realtime, continuous monitoring and quantitative recording of the whole assay process and provides a new insight into the cell-toxicant interaction.


Hypoglycemia and aggression: a review.
The popular notion that a tendency to develop low levels of blood glucose is the cause of a range of behavioral problems is reviewed. It is concluded that it is inappropriate to use the glucose tolerance test as a test of the tendency to develop reactive hypoglycemia. Instead, a meal tolerance test, in which glucose is administered in the presence of fat and protein, should be the method of choice. The use of a meal tolerance test strongly suggests that reactive hypoglycemia rarely results, except in a few exceptional individuals. Three situations are described in which a correlation between a tendency to develop moderately low levels of blood glucose during a glucose tolerance test (not hypoglycemic values) and the tendency to act aggressively have been reported. The significance of these data is unclear but several possible mechanisms by which glucose may influence behavior are discussed.


Autologous bone marrow transplantation in acute lymphoblastic leukemia following in vitro treatment with monoclonal antibodies and with complement: analyses for two cases of failure.
Two children with acute lymphoblastic leukemia (ALL) received autologous bone marrow transplantation (BMT) using remission bone marrow treated in vitro with the monoclonal antibodies, CD24 (BA-1), CD9 (BA-2) and CD10 (BA-3), and with rabbit complement. In one child with second remission ALL, hematopoietic recovery after BMT was prompt but, 81 days after BMT, isolated central nervous system (CNS) relapse occurred. Bone marrow relapse developed three months later, and she died 11 months after BMT. In patients with CNS leukemia prior to BMT, as in the present case, more intensive pretransplant CNS treatment and/or a conditioning regimen may reduce the risk of relapse. In the other patient, with primary refractory ALL in first remission, marrow reconstitution was slower. The patient developed interstitial pneumonitis with pleural effusion, and died 54 days after BMT. No infectious causes could be detected by culture or from serological studies of the pleural effusion. The rationale for applying autologous BMT to children with second remission ALL and first remission refractory ALL is discussed.


Coronary heart disease and its risk factors in a group of 5,000 middle aged men in urban environment.
The results obtained at entry in the subjects included in a pilot study (Institute of Internal Medicine, Bucharest) for the detection and prevention of coronary heart disease and hypertension, are presented. These data are the prevalences of the risk factors of coronary heart disease (high serum cholesterol, hypertension, smoking, overweight, diabetes, nonspecific minor ECG signs, family history), as well as the prevalences of the various forms of coronary heart disease. The study of the frequency distribution of biologic parameters likely to become risk factors showed that in middle aged subjects the upper limit of the normal should be lowered from the 95th percentile to the 76th one.


Vascular endothelial growth factor pathway promotes osseointegration and CD31hiEMCNhi endothelium expansion in a mouse tibial implant model: an animal study.
It is increasingly appreciated that coordinated regulation of angiogenesis and osteogenesis is needed for bone formation. How this regulation is achieved during peri-implant bone healing, such as osseointegration, is largely unclear. This study examined the relationship between angiogenesis and osteogenesis in a unique model of osseointegration of a mouse tibial implant by pharmacologically blocking the vascular endothelial growth factor (VEGF) pathway. An implant was inserted into the right tibia of 16-week-old female C57BL/6 mice (n = 38). Mice received anti-VEGF receptor-1 (VEGFR-1) antibody (25 mg/kg) and VEGF receptor-2 (VEGFR-2) antibody (25 mg/kg; n = 19) or an isotype control antibody (n = 19). Flow cytometric (n = 4/group) and immunofluorescent (n = 3/group) analyses were performed at two weeks post-implantation to detect the distribution and density of CD31hiEMCNhi endothelium. RNA sequencing analysis was performed using sorted CD31hiEMCNhi endothelial cells (n = 2/group). Osteoblast lineage cells expressing osterix (OSX) and osteopontin (OPN) were also detected with immunofluorescence. Mechanical pull-out testing (n = 12/group) was used at four weeks post-implantation to determine the strength of the bone-implant interface. After pull-out testing, the tissue attached to the implant surface was harvested. Whole mount immunofluorescent staining of OSX and OPN was performed to determine the amount of osteoblast lineage cells. Flow cytometry revealed that anti-VEGFR treatment decreased CD31hiEMCNhi vascular endothelium in the peri-implant bone versus controls at two weeks post-implantation. This was confirmed by the decrease of CD31 and endomucin (EMCN) double-positive cells detected with immunofluorescence. In addition, treated mice had more OPN-positive cells in both peri-implant bone and tissue on the implant surface at two weeks and four weeks, respectively. More OSX-positive cells were present in peri-implant bone at two weeks. More importantly, anti-VEGFR treatment decreased the maximum load of pull-out testing compared with the control. VEGF pathway controls the coupling of angiogenesis and osteogenesis in orthopaedic implant osseointegration by affecting the formation of CD31hiEMCNhi endothelium. Cite this article: Bone Joint J 2019;101-B(7 Supple C):108-114.


Specific synthetic peptides for detection of and discrimination between HIV-1 and HIV-2 infection.
A panel of highly purified synthetic oligopeptides representing defined parts of the gag and env proteins of HIV-1 and HIV-2 were used as antigens in ELISA for serodiagnosis of HIV-1 and HIV-2 infection. The analysis included sera from 321 HIV-infected patients and 201 healthy controls from the Ivory Coast, where the prevalence is high for both HIV-1 and HIV-2, and sera from European HIV-1-infected individuals. All sera from HIV-1-infected individuals reacted with a 20 amino acid (a.a.) peptide JB-4c (a.a. 594-613) derived from a highly immunogenic conserved region of the external part of gp41. An equally good response was seen in the HIV-2-infected individuals to a 20 a.a. peptide, JB-16c, from the corresponding part of HIV-2 gp36. Both HIV-1- and HIV-2-seropositive individuals responded well to a peptide, JB-8pc (a.a. 427-448), representing the C-terminal end of the putative CD4-binding site of gp120 of HIV-1. The frequency of reactivity to three selected HIV-1 gag peptides derived from p17 and p15 was 60-70% in both HIV-1 and HIV-2-positive sera. To distinguish between HIV-1 and HIV-2 infection, the sera were titrated against the peptides. Although there was a high degree of cross-reactivity at lower serum dilutions, it was possible to discriminate the infections at higher dilutions to the HIV-1 and HIV-2 gp41/gp36 peptides JB-4c and JB-16c. Analysis of serum reactivity to several selected peptides thus allowed the identification of HIV infection, and the discrimination between HIV-1 and HIV-2.(ABSTRACT TRUNCATED AT 250 WORDS)


A retrospective analysis of hypertension screening at a mass gathering in India: implications for non-communicable disease control strategies.
Cardiovascular disease is the leading case of mortality from non-communicable diseases (NCD) in India. The government's National Programme for Prevention and Control of Cancer, Diabetes, Cardiovascular Diseases and Stroke seeks to increase capacity building, screening, referral and management of NCDs across India, and includes community-based outreach and screening programmes. The government in India routinely provides basic care at religious mass gatherings. However, in 2015, at the Kumbh Mela in Nashik and Trimbakeshwar, the state government extended its services to include a hypertension screening programme. We examine here the value and implications of such opportunistic screening at mass gatherings. At the Kumbh, 5760 persons voluntarily opted for hypertension screening, and received a single blood pressure measurement. In all, 1783 (33.6%) screened positive, of whom, 1580 were previously unaware of their diagnosis. Of the 303 that had previously known hypertension, 240 (79%) were prescribed medications, and 160 were compliant (that is, 52.8% under treatment). Fifty-five (18%) had normal blood pressure readings (BP under control). The data also demonstrated higher prevalence (39%) of hypertension among tobacco users compared to non-users (28%) (P<0.001). Poor recording of phone numbers (0.01%) precluded any phone-based follow-up. The low rates of hypertension awareness, treatment and control underscore the ongoing challenge of both hypertension screening and management in India.


Medicolite-Machine Learning-Based Patient Care Model.
This paper discusses the machine learning effect on healthcare and the development of an application named "Medicolite" in which various modules have been developed for convenience with health-related problems like issues with diet. It also provides online doctor appointments from home and medication through the phone. A healthcare system is "Smart" when it can decide on its own and can prescribe patients life-saving drugs. Machine learning helps in capturing data that are large and contain sensitive information about the patients, so data security is one of the important aspects of this system. It is a health system that uses trending technologies and mobile internet to connect people and healthcare institutions to make them aware of their health condition by intelligently responding to their questions. It perceives information through machine learning and processes this information using cloud computing. With the new technologies, the system decreases the manual intervention in healthcare. Every single piece of information has been saved in the system and the user can access it any time. Furthermore, users can take appointments at any time without standing in a queue. In this paper, the authors proposed a CNN-based classifier. This CNN-based classifier is faster than SVM-based classifier. When these two classifiers are compared based on training and testing sessions, it has been found that the CNN has taken less time (30 seconds) compared to SVM (58 seconds).


Massive sequence perturbation of a small protein.
Most protein topologies rarely occur in nature, thus limiting our ability to extract sequence information that could be used to predict structure, function, and evolutionary constraints on protein folds. In principle, the sequence diversity explored by a given protein topology could be expanded by introducing sequence perturbations and selecting variant proteins that fold correctly. However, our capacity to explore sequence space is intrinsically limited by the enormous number of sequences generated from the 20 amino acids and the limited number of variants likely to fold. Here we sought to test whether the sequence space for naturally existing proteins can be explored by simple, sequential degeneration of a complete set of short sequence segments of a model protein, without long-range covariation. Using the Raf ras binding domain as a model of a small protein capable of autonomous folding, we degenerated 72 of 76 positions of the primary structure for the 20 amino acids in segments of four to seven residues defined by secondary structure and selected the folded species for interaction with h-ras by using an in vivo survival-selection assay. The methodology presented allowed for rigorous statistical analysis and comparison of sequence diversity. The ensemble of sequence variants of Raf ras binding domain obtained have recaptured the diversity observed for the ubiquitin-roll topology. A signature sequence for this fold and the implication of this strategy to protein design and structure prediction are discussed.


Involvement of integrins in fimbriae-mediated binding and invasion by Porphyromonas gingivalis.
Interaction between the major fimbriae of Porphyromonas gingivalis and gingival epithelial cells is important for bacterial adhesion and invasion. In this study, we identified integrins as an epithelial cell cognate receptor for P. gingivalis fimbriae. Immunoprecipitation and direct binding assays revealed a physical association between recombinant fimbrillin and beta1 integrins. In vitro adhesion and invasion assays demonstrated inhibition of binding and invasion of P. gingivalis by beta1 integrin antibodies. In contrast, invasion of a fimbriae-deficient mutant of P. gingivalis was not affected by integrin antibodies. Infection of gingival epithelial cells with wild-type P. gingivalis induced tyrosine phosphorylation of the 68 kDa focal adhesion protein paxillin, whereas the fimbriae-deficient mutant failed to evoke similar changes. Interestingly, activation of paxillin was not accompanied by an increase in the phosphorylation of focal adhesion kinase (FAK). These results provide evidence that P. gingivalis fimbriae promote adhesion to gingival epithelial cells through interaction with beta1 integrins, and this association represents a key step in the induction of the invasive process and subsequent cell responses to P. gingivalis infection.


Proteome profiling of bovine follicular fluid-specific proteins and their effect on in vitro embryo development.
The objective of this study was to determine the effect of bovine follicular fluid proteins (bFF) and their differently charged groups as maturation media supplements for in vitro embryo development. bFF was obtained by aspiration from large healthy follicles (4-10 mm in diameter) and was precipitated by 30-50% (NH4)2SO4. The precipitated protein was fractionated into basic and acidic fractions by ion-exchanger columns. In experiment 1, the oocytes were matured in TCM-199 with 1) FBS+hormones (control) and 2) 10% bFF. The oocyte maturation rate, the development to the blastocyst stage rate and blastocyst cell number were not significantly different between the groups. However, the INFα and IGF-2r expression levels in the 10% bFF were higher than in the control (P<0.05). In experiment 2, the specific charge proteins of bFF (basic and acidic) were also used as media supplements in the maturation medium. The basic fraction had higher oocyte maturation rate and blastocyst cell number when compared with addition of acidic fraction (P<0.05). The expression levels for almost all developmentally important genes in the basic fraction were greater than those in the acidic fraction, particularly INFα (P<0.05). Most of the protein in the basic fraction was associated with the immune response and mRNA processing. In conclusion, supplementation of 10% bFF alone in maturation medium can support oocyte maturation and embryo development. The basic fraction in bFF seemed to have effect on oocyte maturation rate and blastocyst cell number.


Variation of skeletal muscle ultrasound imaging intensity in horses after treadmill exercise: a proof of concept for glycogen content estimation.
Glycogen in skeletal muscle is a major source of energy during exercise and an important determinant of endurance capacity, so that its measurement may provide a meaningful marker of athletes' preparation and a possible predictor of performance, both in humans and in equines. Gold standard of glycogen concentration measurement is the histochemical and biochemical analysis of biopsy-derived muscle tissue, an invasive and potentially injuring procedure. Recently, high-frequency ultrasound (US) technology is being exploited in human sports medicine to estimate muscle glycogen content. Therefore, aim of the present study is to evaluate the feasibility of US assessment of muscle glycogen in equines. US images of gluteus medius (GL) and semitendinosus (ST) muscles were obtained on eight healthy horses (3-10 years) before and after a steady-state exercise on treadmill (velocity: 4.0-12.5 m/s; duration: 2-20 min; heart rate: 137-218 b/min). Average image greyscale intensity was significantly different between GL and ST, both before and after exercise (p < 0.001). Comparing baseline and post-exercise US images, significant increase in greyscale intensity has been observed in ST (p < 0.001), but not in GL (p = 0.129). The volume of the exercise was significantly correlated with exercise-dependent change in image intensity (R2 = 0.891), consistent with a reduction of glycogen muscle stores resulting from aerobic activity. US technique evidences also in horses muscle changes possibly associated to glycogen utilisation during exercise. Present results on a small sample need to be further confirmed and provide preliminary data warranting future validation by direct glycogen measurement through biopsy technique.


Binding of fluorescently labeled cholera toxin subunit B to glycolipids in the human submandibular gland and inhibition of binding by periodate oxidation and by galactose.
FITC-labeled cholera toxin subunit B (CTB) stained the surfaces of cells of mucous acini in the submandibular gland. CTB, also called choleragenoid, binds to the GM1 glycolipid in the cell membrane. The binding in most acini was inhibited by periodic acid oxidation of the sections, while some acini remained unaffected even after increased oxidation. Staining with the subunit was also reduced significantly by adding galactose to the incubation medium. Binding of CTB to cell surfaces apparently requires intact sialic groups on most, but not all, cell surfaces. Oxidation of the sialic acid residues may influence the structure of the sialylated GM1 molecules on the cell surface in different ways. It is possible that both the sialic acid residue and the terminal galactose are oxidized. Alternatively, the sialic acid may be resistant to acid hydrolysis in gangliosides in which the sialic acid is attached to the internal galactose residue linked to GalNAc, as in the GM1 glycolipid. Inhibition of the GM1 receptor binding to cholera toxin has potential for protection of humans against cholera. Galactose and agents that modify sialic acid inhibit the accessibility of the toxin to the GM1 carbohydrate receptor. Human milk contains high levels of sialic acid glycoconjugates that may provide defense mechanisms.


Metronidazole and Demodex folliculorum.
In vitro survival of Demodex folliculorum was tested in the presence of various concentrations of metronidazole (Flagyl). Demodex was found to survive in concentrations of as much as 1 mg/ml. Thus, it seems unlikely that the effect of metronidazole on rosacea is attributable to its direct activity on the mite.


Atherogenic factors and their epigenetic relationships.
Hypercholesterolemia, homocysteine, oxidative stress, and hyperglycemia have been recognized as the major risk factors for atherogenesis. Their impact on the physiology and biochemistry of vascular cells has been widely demonstrated for the last century. However, the recent discovery of the role of epigenetics in human disease has opened up a new field in the study of atherogenic factors. Thus, epigenetic tags in endothelial, smooth muscle, and immune cells seem to be differentially affected by similar atherogenic stimuli. This paper summarizes some recent works on expression of histone-modifying enzymes and DNA methylation directly linked to the presence of risk factors that could lead to the development or prevention of the atherosclerotic process.


Virtual screening for potential inhibitors of Mcl-1 conformations sampled by normal modes, molecular dynamics, and nuclear magnetic resonance.
Myeloid cell leukemia-1 (Mcl-1) is often overexpressed in human cancer and is an important target for developing antineoplastic drugs. In this study, a data set containing 2.3 million lead-like molecules and a data set of all the US Food and Drug Administration (FDA)-approved drugs are virtually screened for potential Mcl-1 ligands using Protein Data Bank (PDB) ID 2MHS. The potential Mcl-1 ligands are evaluated and computationally docked on to three conformation ensembles generated by normal mode analysis (NMA), molecular dynamics (MD), and nuclear magnetic resonance (NMR), respectively. The evaluated potential Mcl-1 ligands are then compared with their clinical use. Remarkably, half of the top 30 potential drugs are used clinically to treat cancer, thus partially validating our virtual screen. The partial validation also favors the idea that the other half of the top 30 potential drugs could be used in the treatment of cancer. The normal mode-, MD-, and NMR-based conformation greatly expand the conformational sampling used herein for in silico identification of potential Mcl-1 inhibitors.


Receptor-Based Discovery of a Plasmalemmal Monoamine Transporter Inhibitor via High Throughput Docking and Pharmacophore Modeling.
Recognition of psychostimulants such as cocaine and the amphetamines by the dopamine transporter (DAT) protein is principally responsible for the euphoria and addiction associated with these drugs. Using as a template the crystal structure of a distantly related bacterial leucine transporter, 3-D DAT computer molecular models have been generated. Ligand docking to such models has revealed potential substrate and inhibitor binding pockets, subsequently confirmed by in vitro pharmacology. An inhibitor pocket defined by the DAT model to be within the "extracellular vestibule", just to the extracellular side of the external gate of the primary substrate pocket, was used for virtual screening of a structural library of compounds. High-throughput docking and application of pharmacophore constraints within this vestibular inhibitor pocket identified a compound structurally dissimilar to the classic monoamine (dopamine, norepinephrine and serotonin) transporter (MAT) inhibitors. The compound displaced binding of radiolabeled cocaine analogs at all three MATs, usually with nanomolar K(i) values and within two fold of cocaine's affinity at the norepinephrine transporter. Although a very weak dopamine uptake inhibitor itself, this compound reduced by three fold the potency of cocaine in inhibiting DAT-mediated cellular uptake of dopamine. To our knowledge, the present findings are the first to successfully employ "receptor-based" computer modeling to identify moderate-to-high affinity MAT ligands. In silico ligand screening using MAT models provides a rapid, low cost discovery process that should accelerate identification of novel ligand scaffolds and provide lead compounds in combating psychostimulant addiction and in treating other monoamine-related CNS diseases.


The proteome of Mycoplasma pneumoniae, a supposedly "simple" cell.
This review covers progress in proteome research on Mycoplasma pneumoniae made over the last 5 years. This bacterium is one of the smallest known self-replicating bacteria. With fewer than 700 proposed proteins, it is well suited to a comprehensive proteome analysis. While all of the proposed genes are transcribed, thus far 620 proteins, about 90% of the predicted proteome, have been identified experimentally. To study the proteome organization of M. pneumoniae, 178 soluble protein complexes were isolated under non-denaturing conditions by tandem affinity chromatography and their composition determined by SDS-PAGE and mass spectrometry. The 62 homomultimeric and 116 heteromultimeric protein complexes could be classified according to 12 different COG functional categories. The complexes interacted with each other to some extent, forming larger assemblies. Protein complexes that were large enough and had specific structures (e.g. ribosomes or DNA-dependent RNA polymerase) were visible and countable in their natural environment by cryo-electron tomography. In addition to characterization of the soluble complexes, the analysis of the Triton X-100 insoluble fraction has a major role in the elucidation of the cytoskeleton-like structure, because by analogy with eukaryotic cells, almost all of the structural proteins involved in its formation, and enriched sub-cellular structures, can be found in this fraction.


Pulsed light beams in vacuum with superluminal and negative group velocities.
We study the group velocity of pulsed light beams in vacuum. Gouy's phase associated with the diffraction of transversally limited pulses can create a strong anomalous dispersion in vacuum leading to highly superluminal and negative group velocities. As a consequence, a focusing pulse can diverge beyond the focus before converging into it. The experimental feasibility is discussed.


Retention of three fissure sealants and a dentin bonding system used as fissure sealant in caries prevention: 12-month follow-up results.
Bonding agents could be used as fissure sealants. This study compares the retention three fissure sealants (Delton, Delton Plus and Concise) and a filled dentin bonding system (Optibond Solo). METHODS. Fifty-six children aged 7-8 years received fissure sealants either in the four permanent first molars, in the four deciduous second molars, or in all eight of these teeth. Every child received a different sealing material in each quadrant on a random basis. Clinical evaluation at 12 months was performed by a single blind examiner, and the retention was classified as either a success (total retention) or a failure (partial retention or not present). There were no statistically significant differences among the four materials in permanent maxillary molars or deciduous molars. In permanent mandibular molars, Optibond Solo showed a lower percentage of retention (40.9%), significantly different (p=0.002) to that of Delton (89.5%), Delton Plus (87.5%) and Concise (76.5%). One bottle dentin bonding system used as a sealant does not improve the retention of conventional fissure sealants. Because of the scarcity of studies on the use of dental adhesives as sealants, further studies are warranted for the final support of that conclusion.


Antineutrophil cytoplasmic antibodies in rheumatoid arthritis. Characterization and clinical correlations.
To study the prevalence, interrelationships, and target antigens of antineutrophil cytoplasmic antibodies (ANCA) in rheumatoid arthritis (RA) and to relate their presence to disease duration and to the occurrence of extraarticular manifestations, including vasculitis. Sera from 94 patients with RA (31 with recent-onset disease, 35 with longstanding disease but without extraarticular manifestations, and 28 with extraarticular disease) were studied for the presence of ANCA by indirect immunofluorescence. All sera were tested by enzyme-linked immunosorbent assay (ELISA) for the presence of antibodies to proteinase 3, myeloperoxidase (MPO), elastase, lactoferrin (LF), and cathepsin G (CG), and by Western blotting for antibodies to neutrophil proteins. Seventy percent of the 94 sera showed staining of the nuclei of ethanol-fixed neutrophils; 32% of the 94 were proven to have ANCA, as manifested by their cytoplasmic staining pattern on paraformaldehyde-fixed neutrophils. In the ELISA, 19 sera reacted with LF, 1 with MPO, and 1 with CG. By Western blotting, 21 sera reacted with LF, and 15 reacted with previously unknown polypeptides (7 sera with a 67/66-kd doublet and 8 with a 63/54-kd doublet). Neither of these antibodies was associated with a particular subset of the disease, but the prevalence of the antibodies tended to increase among patients with longstanding disease. ANCA in RA patients are directed toward diverse cytoplasmic antigens of the neutrophil, in particular, LF and other, not yet fully characterized polypeptides. The antibodies are not a marker for a disease subset, but are probably a corollary of chronic inflammation.


Establishment and Validation of a Nomogram Prediction Model for the Severe Acute Pancreatitis.
Severe acute pancreatitis (SAP) can progress to lung and kidney dysfunction, and blood clotting within 48 hours of its onset, and is associated with a high mortality rate. The aim of this study was to establish a reliable diagnostic prediction model for the early stage of severe pancreatitis. The clinical data of patients diagnosed with acute pancreatitis from October 2017 to June 2022 at the Shangluo Central Hospital were collected. The risk factors were screened by least absolute shrinkage and selection operator (LASSO) regression analysis. A novel nomogram model was then established by multivariable logistic regression analysis. The data of 436 patients with acute pancreatitis, 45 (10.3%) patients had progressed to SAP. Through univariate and LASSO regression analyses, the neutrophils (P <0.001), albumin (P < 0.001), blood glucose (P < 0.001), serum calcium (P < 0.001), serum creatinine (P < 0.001), blood urea nitrogen (P < 0.001) and procalcitonin (P = 0.005) were identified as independent predictive factors for SAP. The nomogram built on the basis of these factors predicted SAP with sensitivity of 0.733, specificity of 0.9, positive predictive value of 0.458 and negative predictive value of 0.967. Furthermore, the concordance index of the nomogram reached 0.889 (95% CI, 0.837-0.941), and the area under the curve (AUC) in receiver operating characteristic curve (ROC) analysis was significantly higher than that of the APACHEII and ABISAP scoring systems. The established model was validated by plotting the clinical decision curve analysis (DCA) and clinical impact curve (CIC). We established a nomogram to predict the progression of early acute pancreatitis to SAP with high discrimination and accuracy.


Platelet-activating factor in septic shock.
Septic shock induced by endotoxins of Gram-negative bacteria, or toxins of Gram-positive bacteria and fungi, deserves particular interest because of its high mortality rate. In experimental animals, treatment with bacterial lipopolysaccharide (endotoxin of Gram-negative bacteria) mimics the symptoms of septic shock. Thus, this treatment has become an important method in animal models of septic shock. Endotoxin induces release of platelet-activating factor and cytokines, such as tumor necrosis factor and interleukins. Platelet-activating factor derived from macrophages, polymorphonuclear leukocytes, and platelets is a potent phospholipid inflammatory mediator that increases cell adhesion and activates endothelial cells by direct effect or through formation of toxic oxygen species and arachidonic acid metabolites, such as thromboxane A2 and leukotriene B4. Platelet-activating factor interacts with cytokines, and this interaction leads to an autocatalytic amplification of inflammatory mediator release. The release of inflammatory mediators by interaction of platelet-activating factor with cytokines is characterized by bell-shaped concentration-effect curves. For example, in a certain concentration range, platelet-activating factor or cytokines induce a mediator release that is proportional to the stimulation. However, over-stimulation may lead to a decrease of mediator release or a prevalence of the release of a single mediator. Down-regulatory processes may be brought about by platelet-activating factor-induced prostacyclin or adenosine release that activates adenylate cyclase and increases intracellular cyclic adenosine 3'5'-monophosphate concentrations. Down-regulation may protect inflammatory and endothelial cells from overstimulation.(ABSTRACT TRUNCATED AT 250 WORDS)


Frequency of Epstein Barr Virus Type 1 Among Nasopharyngeal Carcinomas in Iranian Patients
Background: Around 95% of the world's population are infected with the Epstein-Barr virus (EBV), which can persist latent in B lymphocytes and epithelial cells life-long. EBV has been linked with lymphoid and epithelial cancers and persistence of EBV infection in lymphoid or epithelial cells may result in virus-associated B-cell tumors or nasopharyngeal carcinomas (NPC). This study was conducted to determine the frequency of EBV DNA in nasopharyngeal carcinoma tissue of Iranian patients. Materials and methods: A total of 50 blocks of formalin-fixed paraffin-embedded tissue of NPCs from 38 (76 %) male and 12 (24%) female patients were collected from archives of Ahvaz hospitals. Sections were cut at 5 μm and DNA was extracted for detection of EBV DNA and EBV typing by mested PCR. DNA sequencing was performed to confirm PCR results. The distribution of EBV DNA was compared among WHO histological subtypes of NPC. Results: Some 3 female and 11 (22%) male NPC samples showed positive for EBV DNA type 1, 2/14(22.2%)WHO histological type II and 12/41(29.3%) WHO histological type III. Conclusions: The frequency of EBV DNA among NPCs in Iranian patients was found to be 28%, EBV type I predominating. Both WHO histological type II and III NPC subtypes demonstrated approximately the same detection prevalence.


[Extralevator abdominoperineal excision versus traditional abdominoperineal excision in the treatment of low rectal cancer].
To compare the safety and efficacy between extralevator abdominoperineal excision(ELAPE) and traditional abdominoperineal excision(APE) in patients with low rectal cancer. From January 2011 to December 2013, 61 patients undergoing abdominoperineal excision for low rectal cancer at the Department of Gastrointestinal Surgery, Peking University People's Hospital were enrolled. The safety and efficacy of two procedure groups, ELAPE group (33 patients) and traditional APE group (28 patients) were reviewed retrospectively. Less intraoperative blood loss [(201 ± 147) ml vs. (343 ± 308) ml, P=0.022], shorter exhaust time [(3.8 ± 1.5) d vs. (4.6 ± 1.6) d, P=0.039] and lower perineal wound complication rate (9.1% vs. 25.0%, P=0.042) were observed in the ELAPE group as compared to the traditional APE group. However, longer operation time [(292 ± 46) min vs. (256 ± 28) min, P=0.008] and perineal drainage removal time [(11.1 ± 4.8) d vs. (7.1 ± 2.7) d, P<0.01] were noted in the ELAPE group than those in the traditional APE group. Number of lymph node retrieved and positive lymph node retrieved was not significantly different between two groups. The ELAPE group had lower rate of IOP (intraoperative perforation) (6.1% vs. 17.9%, P=0.055), but no significant difference was found. There were no significant differences in bowel movement, diet restoring time, average perineal drainage, postoperative hospitalization time and removing perineal stitches time between the two groups (all P>0.05). ELAPE possesses less intraoperative blood loss and lower perineal wound complication rate than traditional APE. ELAPE is associated with better safety and efficacy than traditional APE.


[Clinical and experimental investigations of the effect of dopamine on haemodynamics and function of kidney and liver (author's transl)].
Increased incidence of renal insufficiency is observed in severe damage of liver parenchyma such as fulminant hepatitis, decompensated cirrhosis of the liver, septic cholangitis and the different forms of obstructive jaundice. Functional circulatory disturbances of the kidney, especially of the renal cortex, are of importance in the aetiology of this condition. Dopamine, at a dosage as low as 3 gamma/kg/min leads to an improvement in renal blood flow and also to an increase in hepatic blood flow. These observations are of therapeutic importance. Some important circulatory and functional parameters of both these organs, which influence each other under normal and pathological conditions, were studied in the presence of dopamine and the following results were obtained: 1. An investigation of the intrarenal haemodynamics with 133 Xenon in patients with severe cirrhosis of the liver and in patients with obstructive jaundice resulted in an increase of 91% in the mean renal blood flow. The blood flow in the renal cortex increased by 36.2% and in the renal medulla 18.5%, whereas the renal fat tissue showed no change. Compartment I, which was diminished as compared with the control value, also increased. The percentage contribution of the mean renal blood flow and the blood flow of the renal cortex towards the cardiac output was greater under the influence of dopamine; hence a greater part of the cardiac output flows into the kidney under dopamine. 2. The glomerular filtrate and the renal plasma flow increased under dopamine (13.5% and 43.1%, respectively). The increase was greater in compensated than in decompensated cirrhosis. In patients with obstructive jaundice there was a smaller increase in both these parameters than in patients with cirrhosis in the presence of dopamine. No connection was found between the increase in renal plasma flow with dopamine and the blood levels of bilirubin, cholinesterase, GOT and the Normotest. 3. The urinary output of sodium increased by 191.4% with dopamine. Patients with an initial renal plasma flow value of over 300 ml/min had a higher sodium output. These patients also eliminated more sodium under the influence of dopamine than those with an initial renal plasma flow value of under 300 ml/min. 4. Blood flow determinations in the portal vein and the hepatic artery in man, obtained during operation, showed an increase in portal flow of 28.5% and hepatic artery flow of 6.3% in response to dopamine. The percentage contribution of portal blood flow towards the cardiac output increase on dopamine administration. The functional hepatic blood flow, analyzed with 131-J-BSP, did not change. The wedged hepatic vein pressure, which is a good measure of portal pressure, increased on average by only 7% with dopamine at a dosage of 3 gamma/kg/min, but by 20.3% with twice the dosage. Dopamine did not cause a change in hepatic blood volume; hence, blood sequestration in the liver can be excluded in response to the dopamine-evoked increase in portal blood flow. 5...


Protective effect of SIRT3 on acute lung injury by increasing manganese superoxide dismutase-mediated antioxidation.
Prolonged exposure to hyperoxia results in acute lung injury (ALI). Pulmonary damage caused by oxygen toxicity occurs due to the generation of reactive oxygen species and subsequent formation of more potent oxidants. The present study demonstrated that sirtuin 3 (SIRT3) may attenuate hyperoxia‑induced ALI due to its potential antioxidative effect. In the present study, a hyperoxia‑induced acute lung injury mouse model, reverse transcription‑quantitative polymerase chain reaction, western blotting, retroviral mediated gene over‑expression and knockdown assays revealed that the expression of SIRT3 in the lung tissue of mice with hyperoxia‑induced ALI was decreased and overexpression of SIRT3 may significantly reduce hyperoxia‑induced ALI, as reflected by decreases in protein concentration, infiltrated neutrophils in bronchoalveolar lavage (BAL) fluid and wet/dry ratio of lung tissues. Furthermore, overexpression of SIRT3 increased the protein levels and enzymatic activity of manganese superoxide dismutase (MnSOD), and inhibited oxidative stress in the lungs of ALI mice. Additionally, the current study demonstrated that SIRT3 promoted the expression of MnSOD, and this regulation was crucial for the protective effect of SIRT3 on hyperoxia‑induced ALI. In summary, the results of the current study indicated that SIRT3 overexpression may effectively ameliorate hyperoxia‑induced ALI in mice, which indicates a potential application for SIRT3‑based gene therapy to treat clinical adult respiratory distress syndrome.


Obsessive-compulsive disorder: defining the phenotype.
Characterizing the obsessive-compulsive disorder (OCD) phenotype is important for treatment and etiologic studies. This article describes a family-study approach for identifying the spectrum of conditions related to OCD and subtyping OCD cases into homogeneous subtypes.


Rasch Analysis of the Teachers' Knowledge and Use of Data and Assessment (tKUDA) Measure.
Teachers are expected to use data and assessments to drive their instruction. This is accomplished at a classroom level via the assessment process. The teachers Knowledge and Use of Data and Assessment (tKUDA) measure was created to capture teachers' knowledge and use of this assessment process. This paper explores the measure's utility using Rasch analysis. Evidence of reliability and validity was seen for both knowledge and use factors. Scale was used as expected and item analyses demonstrates good spread with a few items identified for future revision. Item difficulty and results are connected back to literature. Findings support use of this measure to identify teachers' knowledge and use of data and assessment in classroom practice.