Rho GTPases as targets of bacterial protein toxins.
Several bacterial toxins target Rho GTPases, which constitute molecular switches in several signaling processes and master regulators of the actin cytoskeleton. The biological activities of Rho GTPases are blocked by C3-like transferases, which ADP-ribosylate Rho at Asn41, but not Rac or Cdc42. Large clostridial cytotoxins (e. g., Clostridium difficile toxin A and B) glucosylate Rho GTPases at Thr37 (Rho) or Thr35 (Rac/Cdc42), thereby inhibiting Rho functions by preventing effector coupling. The 'injected' toxins ExoS, YopE and SptP from Pseudomonas aeruginosa, Yersinia and Salmonella ssp., respectively, which are transferred into the eukaryotic target cells by the type-III secretion system, inhibit Rho functions by acting as Rho GAP proteins. Rho GTPases are activated by the cytotoxic necrotizing factors CNF1 and CNF2 from Escherichia coli and by the dermonecrotizing toxin DNT from B. bronchiseptica. These toxins deamidate/transglutaminate Gln63 of Rho to block the intrinsic and GAP-stimulated GTP hydrolysis, thereby constitutively activating the GTPases. Rho GTPases are also activated by SopE, a type-III system injected protein from Salmonella ssp., that acts as a GEF protein.


Drugs producing vitamin deficiencies.
Many drugs produce vitamin deficiencies. They belong to the most important and common therapeutical classes: analgesics, antianemics, antibacterial and antiblastic agents, antibiotics, antidiabetics, antimalarials, antiphlogistics, antipyretics, diuretics, laxatives and purgatives, tranquilizers and anticonvulsives, radiomimetics, hormones and vitamins themselves. The vitamin deprivation processes may be produced by a variety of mechanisms and may involve all vitamins. Recent experiments indicate that there is a competition for binding sites on proteins between vitamin C and salicylate and between dicoumarol and vitamin K. Usually a drug exerts a "devitaminizing" action with respect to only one vitamin. However there are examples of multiple vitamin deficiencies induced by a single drug, like salicylate which deprives the organism of vitamins C, K and pantothenate. These deficiencies may develop either all at the same time or successively. A direct and concomitant vitamin depriving action occurs when an antibiotic blocks the production of vitamins by the enteric flora. A different mode of action occurs in the drug induced folic acid deficiency, which in turn induces a deficiency of vitamin B12. It has been reported that a vitamin deficiency may result from intake of high pharmacological doses of other vitamins. These data need confirmation in patients treated with high doses of nicotinic acid. The drug induced vitamin deficiencies are studied with the same methodology employed for avitaminoses in general; hence they can be diagnosed using the same criteria.


Pseudoephedrine hydrochloride sustained-release pellets prepared by a combination of hot-melt subcoating and polymer coating.
Pseudoephedrine hydrochloride is an active very highly water soluble substance. In order to control release of a drug with this property, we developed the application of a combination of hot-melt subcoating and polymer coating was developed. The main objective was to investigate the influence of this combination on the release of highly water soluble drug and how it works. Hot-melt subcoating was achieved by using a coating pan. Subsequently, the outer polymer coating was prepared by fluidized bed, and the drug release was determined by high-performance liquid chromatograph (HPLC) method. Hot-melt subcoating can form a barrier between the drug-loaded pellets and the polymer coating layer, which prevents migration of the drug during film application. Consequently, the level of polymer coating can be reduced significantly, and the effectiveness of the polymer coating increased. In this study, the release profile of pellets with a 10% hot-melt subcoating and 5% polymer coating weight gain met the dissolution requirement of USP29 for pseudoephedrine hydrochloride extended-release capsules. Compared with pellets only polymer coated (10% level), the polymer coating level of pellets prepared by this technology was reduced by half due to hot-melt subcoating. By means of this hot-melt subcoating and polymer coating, sustained-release pellets containing pseudoephedrine hydrochloride were successfully prepared.


Sagittal joint instability in the cranial cruciate ligament insufficient canine stifle. Caudal slippage of the femur and not cranial tibial subluxation.
This in vivo study qualitatively describes the sagittal motion pattern of the cranial cruciate ligament (CrCL) insufficient canine stifle in operated and unoperated joints with cranio-caudal laxity on palpation. Sagittal stifle kinematics were recorded in vivo in dogs (> 15 kg BW) with unilateral (n = 7) or bilateral (n = 6) complete CrCL rupture and positive cranial drawer test as well as two sound control dogs using uniplanar fluoroscopic kinematography with the dogs walking on a treadmill. Stifle stability and sagittal motion pattern of the femur and the tibia were determined by visual inspection of the fluoroscopic video sequences. Control dogs showed no cranio-caudal instability, identical to the contralateral stifles of the dogs with unilateral rupture. All unoperated stifles with CrCL rupture (n = 6) showed caudal slippage of the femur at the beginning of the stance phase. Of the 13 operated stifles (TightRope: n = 1, tibial tuberosity advancement, TTA: n = 6, tibial plateau leveling osteotomy, TPLO: n = 5, cranial closing wedge osteotomy, CCWO: n = 1) nine were unstable, showing the same motion pattern as the unoperated stifles. In the CrCL insufficient stifle with in vivo cranio-caudal instability caudal slippage of the distal femur at tow touch is the predominant motion pattern. The discrepancy between in vivo motion pattern and in vitro simulation of CrCL insufficiency in which cranial tibial subluxation is the predominant sagittal motion pattern warrants further studies.


Oxygen inhalation improves survival time of mice with acute intra-abdominal hypertension and protects liver cells.
The objectives of this study were to establish a mouse model of sustainable, stable acute intra-abdominal hypertension (IAH), seeking to observe the survival time and liver functions at different intra-abdominal pressures (IAP), and to investigate changes after oxygen therapy. Sixty Kunming mice were assigned to 4 groups with an average of 15, 20, 30, or 40 cmH(2)O IAP. The 40 cmH(2)O group seemed to be appropriate for follow-up experiments. The 45 mice added to the cohort were assigned into 3 groups administered an average of 50%, 80%, and 100% O(2), respectively. Liver and blood samples were used to compare the rates of apoptosis using the TUNEL assay as well as alanine aminotransferase (ALT), aspartate aminotransferase (AST); Caspase-3, 9, MDA, and SOD concentrations. Most animals in the 15 and 20 cmH(2)O groups survived 8 hours. The average survival for the 30 and 40 cmH(2)O groups were 4.54 ± 0.54 hours and 2.04 ± 0.44 hours, respectively (P < .01). As the oxygen concentration increased, the survival time was prolonged among the 40 cmH(2)O IAP group (P < .01), and the member of apoptotic hepatic cells decreased (P < .01), with a concomittent decrease in caspase 3 and 9 as well as malondialdehyde, although superoxide dismutase showed the opposite results. The present work using a mouse model for acute IAH showed oxygen inhalation to improve host survival and protect liver cells.


[Chief types of placental trophism].
An attempt is made to represent a theory on two types of placental throphism. The first type--nutrition is mainly performed at the expense of maternal proteins, which in the placental chorion undergo proteolysis with formation of polypeptides and amino acids; embryospecific proteins are formed in the liver of the embryo. This type is characteristic for the animals with epithelio-chorionic acid and partly with desmo-chorionic placentas. The second type is characterized by resorption of amino acids from the maternal organism, from the latter embryospecific proteins are synthesized in the chorion. Thus, placental nutrition is specific for man and animals with hemochorionic and, partly, with endothelio-chorionic placentas.


Three-dimensional surface mapping of the caudate nucleus in late-life depression.
To compare the volumes of the caudate nucleus, using traditional volumetry and a three-dimensional brain mapping technique, in a group of individuals with late-life depression and a group of age- and education-equated nondepressed comparison subjects. Cross-sectional. University Medical Center. Twenty-three nondemented subjects with late-life depression and 15 age- and education-equated elderly comparison subjects (depressed mean years of age: 70.5 +/- 5.7 SD, comparison subjects = 69.9 years +/- 6.4) with no history of psychiatric or neurologic disease. Structural magnetic resonance imaging. Three-dimensional (3-D) surface models were created from manually traced outlines of the caudate nucleus from spoiled gradient echo images. Models were geometrically averaged across subjects and statistical maps created to localize any regional volume differences between groups. Relative to comparison subjects, depressed subjects had significantly lower mean volumes for both the left (p = 0.029) and right (p = 0.052) caudate nucleus as well as total caudate volume (p = 0.032). Total volumes were 13.1% less in the depressed group (13.5% on the left and 12.6% on the right). 3-D maps further localized these reductions to the caudate head. Volume reductions were correlated with depression severity, as measured by the 17-item Hamilton Depression Rating Scale. Late-life depression is associated with left and right caudate nucleus reduction especially in anterior portions. Among depressed subjects, greater caudate reduction was associated with more severe depression. These results are consistent with growing evidence that the anterior caudate nucleus, especially the head, may be structurally and functionally abnormal in affective disorders.


Positive and negative emotionality: trajectories across six years and relations with social competence.
The goals of the present study were to examine (1) the mean-level stability and differential stability of children's positive emotional intensity, negative emotional intensity, expressivity, and social competence from early elementary school-aged to early adolescence, and (2) the associations between the trajectories of children's emotionality and social functioning. Using four waves of longitudinal data (with assessments 2 years apart), parents and teachers of children (199 kindergarten through third grade children at the first assessment) rated children's emotion-related responding and social competence. For all constructs, there was evidence of mean-level decline with age and stability in individual differences in rank ordering. Based on age-centered growth-to-growth curve analyses, the results indicated that children who had a higher initial status on positive emotional intensity, negative emotional intensity, and expressivity had a steeper decline in their social skills across time. These findings provide insight into the stability and association of emotion-related constructs to social competence across the elementary and middle school years.


FIONA1-mediated methylation of the 3'UTR of FLC affects FLC transcript levels and flowering in Arabidopsis.
Adenosine bases of RNA can be transiently modified by the deposition of a methyl-group to form N6-methyladenosine (m6A). This adenosine-methylation is an ancient process and the enzymes involved are evolutionary highly conserved. A genetic screen designed to identify suppressors of late flowering transgenic Arabidopsis plants overexpressing the miP1a microProtein yielded a new allele of the FIONA1 (FIO1) m6A-methyltransferase. To characterize the early flowering phenotype of fio1 mutant plants we employed an integrative approach of mRNA-seq, Nanopore direct RNA-sequencing and meRIP-seq to identify differentially expressed transcripts as well as differentially methylated RNAs. We provide evidence that FIO1 is the elusive methyltransferase responsible for the 3'-end methylation of the FLOWERING LOCUS C (FLC) transcript. Furthermore, our genetic and biochemical data suggest that 3'-methylation stabilizes FLC mRNAs and non-methylated FLC is a target for rapid degradation.


Fraught relations in the letters of Laura Cereta: marriage, friendship, and humanist epistolarity.
Laura Cereta is unique among Quattrocento female humanists in directly addressing the position of women as wives and as friends in her substantial corpus of erudite Latin epistolary prose. Questioning the ideals that governed intellectual, social, and personal expectations of matrimony, Cereta's letters reflect her self-consciously double status as humanist and spouse. Her fierce critique of marriage as a site of female oppression and complicity implies an alternative that requires of humanists, husbands, and wives a radical rethinking of marriage in terms of friendship, as well as of the very project of humanist epistolarity.


Characterization of Cestrum yellow leaf curling virus: a new member of the family Caulimoviridae.
Cestrum yellow leaf curling virus (CmYLCV) has been characterized as the aetiological agent of the Cestrum parqui mosaic disease. The virus genome was cloned and the clone was proven to be infectious to C. parqui. The presence of typical viroplasms in virus-infected plant tissue and the information obtained from the complete genomic sequence confirmed CmYLCV as a member of the Caulimoviridae family. All characteristic domains conserved in plant pararetroviruses were found in CmYLCV. Its genome is 8253 bp long and contains seven open reading frames (ORFs). Phylogenetic analysis of the relationships with other members of the Caulimoviridae revealed that CmYLCV is closely related to the Soybean chlorotic mottle virus (SbCMV)-like genus and particularly to SbCMV. However, in contrast to the other members of this genus, the primer-binding site is located in the intercistronic region following ORF Ib rather than within this ORF, and an ORF corresponding to ORF VII is missing.


Blood pressure measurement in different outpatient clinics.
The objective of this study is to investigate how blood pressure is measured in different outpatient clinics. This is a retrospective study performed with 1,000 patients, randomly selected among patients followed in different outpatient clinics in Fortaleza, Brazil. Among the 1,000 patients, 962 had available records. The older the patient, the greater the possibility of having had blood pressure measured. In subsequent visits, blood pressure was measured more often than in the first visits. Blood pressure was measured more often among patients with chronic disease. The specialty with the highest proportion of blood pressure measurement was cardiology (85%). At the surgery, dermatology and traumatology clinics, blood pressure was measured in less than 5% of visits, and at the ophthalmology, sexually transmitted diseases, psychiatry, proctology and otolaryngology clinics, blood pressure was not measured in any visit. Cardiology, endocrinology, nephrology, geriatrics and internal medicine were the specialties where blood pressure was more frequently measured. It is important to measure blood pressure in any medical specialty in order to give early diagnosis of hypertension and provide adequate blood pressure control.


A new osteotomy for cubitus varus.
From 1974 to 1986, a step-cut technique of distal humerus valgus osteotomy using one cortical screw for fixation was used to correct cubitus varus deformity in 11 patients. The results were graded as excellent, eight patients, good, two, and poor, one. The poor result was secondary to persistent varus. The average humeral-elbow-wrist angle in the ten patients with valgus correction roentgenographically measured 9.3 degrees. The average amount of correction was 28.4 degrees. All patients retained their preoperative level of elbow motion. There were no radial or ulnar nerve injuries, nonunions, infections, or hypertrophic scars. The osteotomy requires careful preoperative planning and special attention to surgical detail. Large amounts of deformity may be corrected safely with a low complication rate.


Survey of simple sequence repeats in completed fungal genomes.
The use of simple sequence repeats or microsatellites as genetic markers has become very popular because of their abundance and length variation between different individuals. SSRs are tandem repeat units of 1 to 6 base pairs that are found abundantly in many prokaryotic and eukaryotic genomes. This is the first study examining and comparing SSRs in completely sequenced fungal genomes. We analyzed and compared the occurrences, relative abundance, relative density, most common, and longest SSRs in nine taxonomically different fungal species: Aspergillus nidulans, Cryptococcus neoformans, Encephalitozoon cuniculi, Fusarium graminearum, Magnaporthe grisea, Neurospora crassa, Saccharomyces cerevisiae, Schizosaccharomyces pombe, and Ustilago maydis. Our analysis revealed that, in all of the genomes studied, the occurrence, abundance, and relative density of SSRs varied and was not influenced by the genome sizes. No correlation between relative abundance and the genome sizes was observed, but it was shown that N. crassa, the largest genome analyzed had the highest relative abundance of SSRs. In most genomes, mononucleotide, dinucleotide, and trinucleotide repeats were more abundant than the longer repeated SSRs. Generally, in each organism, the occurrence, relative abundance, and relative density of SSRs decreased as the repeat unit increased. Furthermore, each organism had its own common and longest SSRs. Our analysis showed that the relative abundance of SSRs in fungi is low compared with the human genome and that longer SSRs in fungi are rare. In addition to providing new information concerning the abundance of SSRs for each of these fungi, the results provide a general source of molecular markers that could be useful for a variety of applications such as population genetics and strain identification of fungal organisms.


The Special Measures for Quality and Challenged Provider Regimes in the English NHS: A Rapid Evaluation of a National Improvement Initiative for Failing Healthcare Organisations.
There is limited knowledge about interventions used for the improvement of low-performing healthcare organisations and their unintended consequences. Our evaluation sought to understand how healthcare organisations in the National Health Service (NHS) in England responded to a national improvement initiative (the Special Measures for Quality [SMQ] and challenged provider [CP] regimes) and its perceived impact on achieving quality improvements (QIs). Our evaluation included national-level interviews with key stakeholders involved in the delivery of SMQ (n=6); documentary analysis (n=20); and a qualitative study based on interviews (n=60), observations (n=8) and documentary analysis (n=291) in eight NHS case study sites. The analysis was informed by literature on failure, turnaround and QI in organisations in the public sector. At the policy level, SMQ/CP regimes were intended to be "support" programmes, but perceptions of the interventions at hospital level were mixed. The SMQ/CP regimes tended to consider failure at an organisational level and turnaround was visualised as a linear process. There was a negative emotional impact reported by staff, especially in the short-term. Key drivers of change included: engaged senior leadership teams, strong clinical input and supportive external partnerships within local health systems. Trusts focused efforts to improve across multiple domains with particular investment in improving overall staff engagement, developing an open, listening organisational culture and better governance to ensure clinical safety and reporting. Organisational improvement in healthcare requires substantial time to embed and requires investment in staff to drive change and cultivate QI capabilities at different tiers. The time this takes may be underestimated by external 'turn-around' interventions and performance regimes designed to improve quality in the short-term and which come at an emotional cost for staff. Shifting an improvement focus to the health system or regional level may promote sustainable improvement across multiple organisations over the long-term.


Prolonged recovery of memory functioning after mild traumatic brain injury in adolescent athletes.
The current body of sports-related concussion literature is hampered by a lack of research conducted in high school athletes. Accordingly, the authors sought to examine the neuropsychological deficits and recovery patterns after concussive injuries in this population. Participants included 419 male and female athletes with a mean age of 15.69 years who underwent baseline testing of their neuropsychological functioning prior to their sports season. Fourteen participants sustained an in-season concussion and were serially reassessed at ~2.5, 6, and 10 days postinjury. Fourteen uninjured matched control participants were also reassessed at the end of the school year. Individuals who sustained in-season concussions demonstrated impairments in reaction time, processing speed, and had delayed memory functioning. Although reaction time and processing speed deficits returned to baseline levels by ~6 days postinjury, participants continued to show memory impairments up to 7 days postinjury. Memory impairments were found to resolve by Day 10, however. The results of the present study suggest that high school athletes demonstrate prolonged memory dysfunction compared with college athletes, and should therefore be treated more conservatively.


Influence of Loading Conditions and Temperature on Static Friction and Contact Aging of Hydrogels with Modulated Microstructures.
Biological tribosystems enable diverse functions of the human body by maintaining extremely low coefficients of friction via hydrogel-like surface layers and a water-based lubricant. Although stiction has been proposed as a precursor to damage, there is still a lack of knowledge about its origin and its relation to the hydrogel's microstructure, which impairs the design of soft matter as replacement biomaterials. In this work, the static friction of poly(acrylamide) hydrogels with modulated composition was investigated by colloidal probe lateral force microscopy as a function of load, temperature, and loading time. Temperature-dependent studies enable to build a phase diagram for hydrogel's static friction, which explains stiction via (polymer) viscoelastic and poroelastic relaxation, and a subtle transition from solid- to liquid-like interfacial behavior. At room temperature, the static friction increases with loading time, a phenomenon called contact aging, which stems from the adhesion of the polymer to the colloid and from the drainage-induced increase in contact area. Contact aging is shown to gradually vanish with increase in temperature, but this behavior strongly depends on the hydrogel's composition. This work scrutinizes the relation between the microstructure of hydrogel-like soft matter and interfacial behavior, with implications for diverse areas of inquiry, not only in biolubrication and biomedical applications but also in soft robotics and microelectromechanical devices, where the processes occurring at the migrating hydrogel interface are of relevance. The results support that modulating both the hydrogel's mesh size and the structure of the near-surface region is a means to control static friction and adhesion. This conceptual framework for static friction will foster further understanding of the wear of hydrogel-like materials.


Long-read assembly and comparative evidence-based reanalysis of Cryptosporidium genome sequences reveal expanded transporter repertoire and duplication of entire chromosome ends including subtelomeric regions.
Cryptosporidiosis is a leading cause of waterborne diarrheal disease globally and an important contributor to mortality in infants and the immunosuppressed. Despite its importance, the Cryptosporidium community has only had access to a good, but incomplete, Cryptosporidium parvum IOWA reference genome sequence. Incomplete reference sequences hamper annotation, experimental design, and interpretation. We have generated a new C. parvum IOWA genome assembly supported by Pacific Biosciences (PacBio) and Oxford Nanopore long-read technologies and a new comparative and consistent genome annotation for three closely related species: C. parvum, Cryptosporidium hominis, and Cryptosporidium tyzzeri We made 1926 C. parvum annotation updates based on experimental evidence. They include new transporters, ncRNAs, introns, and altered gene structures. The new assembly and annotation revealed a complete Dnmt2 methylase ortholog. Comparative annotation between C. parvum, C. hominis, and C. tyzzeri revealed that most "missing" orthologs are found, suggesting that the biological differences between the species must result from gene copy number variation, differences in gene regulation, and single-nucleotide variants (SNVs). Using the new assembly and annotation as reference, 190 genes are identified as evolving under positive selection, including many not detected previously. The new C. parvum IOWA reference genome assembly is larger, gap free, and lacks ambiguous bases. This chromosomal assembly recovers all 16 chromosome ends, 13 of which are contiguously assembled. The three remaining chromosome ends are provisionally placed. These ends represent duplication of entire chromosome ends including subtelomeric regions revealing a new level of genome plasticity that will both inform and impact future research.


Antiproliferative activity of Ontario grown onions against colorectal adenocarcinoma cells.
Cancer is the leading cause of mortality in Canada and other industrialized nations; the development of new/improved cancer therapies is desperately needed and continues to be a major focus of cancer research. Flavonoids, which are found in high levels in onions, have been shown to exert antiproliferative and potentially anti-cancer activities. To test their therapeutic potential, we assessed the antiproliferative, cytotoxic, apoptosis-inducing, and anti-migratory activities of five onion varieties grown in Ontario against human adenocarcinoma (Caco-2) cells. The properties of onion extracts were compared to pure extracts of flavonoids known to exhibit antiproliferative effects (quercetin, myricetin, and kaempferol). We compared more than one variety of onion, as agronomic and genetic factors influence the composition, as well as the quality of phytochemicals (e.g. flavonoids) in plant cultivars. We found that all onion varieties exhibited antiproliferative activity similar to purified flavonoids. The cytotoxic effects of the Stanley and Fortress onion varieties were strongest among the selected cultivars, as determined via lactate dehydrogenase (LDH) assays, while Safrane extracts showed the weakest activity. The Stanley and Lasalle cultivar extracts also had strong anti-migratory effects. Altogether these onion extracts may contain one or more compounds that may be effective anti-cancer therapies, while the Stanley extract showed the most comprehensive biological activities against Caco-2 cells.


[Epidermoid carcinoma of the scrotum].
To analyze the therapeutic approach in epidermoid or squamous cell carcinoma of the scrotum, a rare tumor that is of interest for clinical and historical reasons. A case of epidermoid carcinoma of the scrotum is described. The patient had undergone urethroplasty in 1972, using preputial free graft, according to the technique of Ben Johnson. Right scrotectomy and orchidectomy were performed and partial urethrectomy with inguinal lymphadenectomy 6 weeks later. At 24 months' follow-up, the patient has no tumor recurrence. Malignant tumors of the scrotum are rare. Squamous cell carcinoma is the most frequent tumor type. Radical surgery with inguinal lymphadenectomy is the treatment of choice. It must be distinguished from verrucous carcinoma, a tumor that requires a different therapeutic approach and for which radical surgery is not indicated.


Chlorido-containing ruthenium(II) and iridium(III) complexes as antimicrobial agents.
A series of polypyridyl-ruthenium(II) and -iridium(III) complexes that contain labile chlorido ligands, [{M(tpy)Cl}(2){μ-bb(n)}](2/4+) {Cl-Mbb(n); where M = Ru or Ir; tpy = 2,2':6',2''-terpyridine; and bb(n) = bis[4(4'-methyl-2,2'-bipyridyl)]-1,n-alkane (n = 7, 12 or 16)} have been synthesised and their potential as antimicrobial agents examined. The minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of the series of metal complexes against four strains of bacteria - Gram positive Staphylococcus aureus (S. aureus) and methicillin-resistant S. aureus (MRSA), and Gram negative Escherichia coli (E. coli) and Pseudomonas aeruginosa (P. aeruginosa) - have been determined. All the ruthenium complexes were highly active and bactericidal. In particular, the Cl-Rubb(12) complex showed excellent activity against all bacterial cell lines with MIC values of 1 μg mL(-1) against the Gram positive bacteria and 2 and 8 μg mL(-1) against E. coli and P. aeruginosa, respectively. The corresponding iridium(III) complexes also showed significant antimicrobial activity in terms of MIC values; however and surprisingly, the iridium complexes were bacteriostatic rather than bactericidal. The inert iridium(III) complex, [{Ir(phen)(2)}(2){μ-bb(12)}](6+) {where phen = 1,10-phenanthroline) exhibited no antimicrobial activity, suggesting that it could not cross the bacterial membrane. The mononuclear model complex, [Ir(tpy)(Me(2)bpy)Cl]Cl(2) (where Me(2)bpy = 4,4'-dimethyl-2,2'-bipyridine), was found to aquate very rapidly, with the pK(a) of the iridium-bound water in the corresponding aqua complex determined to be 6.0. This suggests the dinuclear complexes [Ir(tpy)Cl}(2){μ-bb(n)}](4+) aquate and deprotonate rapidly and enter the bacterial cells as 4+ charged hydroxo species.


[Effects of an alternative program to promote healthy eating in school canteen].
The objective of this study was to determine the effects on school feeding habits in an educational intervention alternative to the conventional model of feeding promotion to a center with school canteens and ecological garden project. Cross-sectional and comparative descriptive observational study of 111 boys and girls between 9 and 12 years old, in the 2013-15 period. A school with alternative intervention was compared with another one that only included contents of the Infant and Primary Education curriculum, by means of evaluation of adherence to the mediterranean diet of the students with a validated Kidmed questionnaire. The intervention center presented greater adherence to the mediterranean diet than the control center. In the intervention center, the percentage of students with optimal diet (64.52%) was higher than those who needed to improve their diet (33.87%), contrary to what occurs in control center (42.86% and 51.02% respectively). The students of the intervention center presented a higher percentage of affirmative answers in Kidmed test. The program of promotion of healthy alternative food of the intervention center could be effective for the acquisition of a diet of greater adherence to the mediterranean diet.


Pharmacology of emesis and gastrointestinal motility: implications for migraine.
This paper outlines the physiopharmacology of emesis and its gastrointestinal motor correlates in relation to the possible treatment strategies of a migraine attack. The gastrointestinal motor correlates of vomiting consist of a stereotyped motor response to diverse stimuli and basically include gastric relaxation and a series of events (intestinal motor inhibition and disruption of slow wave activity) in preparation for the occurrence of a retrograde giant contraction that propagates orally and empties intestinal contents into the stomach. These motor correlates always accompany vomiting, but may also appear as an independent phenomenon. Antidopaminergic agents, such as metoclopramide and domperidone, have antiemetic/prokinetic properties, block the gastrointestinal motor correlates of vomiting and may speed up absorption of antimigraine agents. The use of 5-HT4 receptor agonists may be an alternative strategy to achieve prokinesia during a migraine attack, although this issue still needs to be fully addressed by controlled studies. Sumatriptan was found to induce gastric relaxation and delay gastric emptying, but the impact of these effects during a migraine attack remains to be determined.


Structural Contour Map of the Iota Carbonic Anhydrase from the Diatom Thalassiosira pseudonana Using a Multiprong Approach.
Carbonic anhydrases (CAs) are a family of ubiquitous enzymes that catalyze the interconversion of CO2 and HCO3-. The "iota" class (ι-CA) was first found in the marine diatom Thalassiosira pseudonana (tpι-CA) and is widespread among photosynthetic microalgae and prokaryotes. The ι-CA has a domain COG4875 (or COG4337) that can be repeated from one to several times and resembles a calcium-calmodulin protein kinase II association domain (CaMKII-AD). The crystal structure of this domain in the ι-CA from a cyanobacterium and a chlorarachniophyte has been recently determined. However, the three-dimensional organization of the four domain-containing tpι-CA is unknown. Using biophysical techniques and 3-D modeling, we show that the homotetrameric tpι-CA in solution has a flat "drone-like" shape with a core formed by the association of the first two domains of each monomer, and four protruding arms formed by domains 3 and 4. We also observe that the short linker between domains 3 and 4 in each monomer confers high flexibility, allowing for different conformations to be adopted. We propose the possible 3-D structure of a truncated tpι-CA containing fewer domain repeats using experimental data and discuss the implications of this atypical shape on the activity and metal coordination of the ι-CA.


Association of Proton Pump Inhibitor Use With Serum Biomarkers of Inflammation, Insulin Resistance, Cardiovascular Risk, and Renal Function.
Proton pump inhibitor (PPI) use has been associated with cardiovascular disease, chronic kidney disease, and dementia. Prior studies did not account for key confounders and little is known about the association of PPIs with serum biomarkers of inflammation, insulin resistance, cardiovascular risk, and renal function. Our aims were to investigate differences in these biomarkers between PPI users and nonusers. Our data are from the National Health and Nutrition Examination Survey (NHANES), a complex cross-sectional multistage probability sample of the US civilian population. We used data on 5189 eligible adults aged 18 to 85 years. Appropriate survey commands were used and potential confounding variables (including BMI, duration of PPI use, use of other non-PPI medications, and health behaviors) were included in multivariable regression models assessing biomarker outcomes. PPI use was associated with differences in mean (±SE) fasting low-density lipoprotein (LDL) (by 11.7±3.7 mg/dL; P=0.006), and apolipoprotein B (by 7.6±2.6 mg/dL; P=0.01). PPI use was not associated with significant differences in total cholesterol (P=0.13), high-density lipoprotein (P=0.27), triglycerides (P=0.70), c-reactive protein (P=0.52), the homeostatic model assessment-insulin resistance (P=0.48), hemoglobin A1c (P=0.39), or homocysteine (P=0.87). PPI use was associated with a decrease in blood urea nitrogen (by 1.0±0.3 mg/dL; P=0.008) but not creatinine (P=0.38) or uric acid (P=0.34). PPI was not associated with clinically significant differences in serum biomarkers of inflammation, insulin resistance, cardiovascular risk, and renal function. Rather, increasing BMI was strongly associated with PPI use and clinically significant differences in these biomarkers.


Regulation of gelatinase B production in corneal cells is independent of autocrine IL-1alpha.
The matrix metalloproteinase gelatinase B is synthesized by cells at the leading edge of the corneal epithelium migrating to heal a wound. Recent data from the authors' laboratory suggest that excessive synthesis contributes to repair defects. The goal of the study reported here was to investigate mechanisms controlling gelatinase B production by corneal epithelial cells. Freshly isolated cultures of corneal epithelial cells and early passage stromal fibroblasts from rabbit were used for these studies. In a previous study, it was found that the cytokine interleukin (IL)-1alpha is released into the culture medium of corneal epithelial cells more efficiently when they are plated at low density with limited cell-cell contact than when plated at high density. In this study, we show that production of gelatinase B by these cells is similarly affected by cell plating density. However, it is further demonstrated that these two events are not dependent on one another but occur in parallel: IL-1alpha does not regulate gelatinase B production (synthesis), nor was there evidence that any other secreted autocrine cytokine acts as mediator. Instead, our data suggest that gelatinase B production is downregulated directly by high cell density and indicate a connection to the level of protein kinase C activity. Nevertheless, the anticancer agent suramin, which blocks collagenase synthesis by interfering with autocrine cytokine-receptor interactions, still inhibits synthesis of gelatinase B. Unlike collagenase synthesis by corneal stromal fibroblasts, production (synthesis) of gelatinase B does not appear to be controlled by secreted autocrine cytokines but can still be inhibited by suramin. Suramin may make an effective therapeutic agent for controlling pathologic overproduction of gelatinase B in corneal ulcers.


Diabetes mellitus and pregnancy.
Diabetes mellitus is the most common medical complication of pregnancy and it carries a significant risk to the foetus and the mother. Congenital malformations and perinatal morbidity remain common compared with the offspring of non diabetic pregnancies. Diabetic mothers are at risk of progression of microvascular diabetic complications as well as early pregnancy loss, pre-eclampsia, polyhydramnios and premature labour. Glycaemic control before and during pregnancy is critical and the benefit may result in a viable, healthy off spring. Gestational diabetes mellitus (GDM) which manifests for the first time during pregnancy is common and on the increase, its proper management will reduce the risk of neonatal macrosomia and hypoglycaemia. Post-partum evaluation of glucose tolerance and appropriate counselling in women with GDM may help decrease the high risk of subsequent type 2 diabetes in the longterm.This article will briefly review the changes in the carbohydrate metabolism that characterise normal pregnancy and will focus on a practical approach to the care of patients with pre-existing diabetes as well as GDM.


Isolation and genetic analysis of amber uvrA and uvrB mutants.
Genetic properties of amber uvrA and uvrB mutants of Escherichia coli K-12 are described. The isolation of three amber uvrA and two amber uvrB mutants indicates that the products of these genes are proteins.


Association of Kidney Function Measures With Signs of Neurodegeneration and Small Vessel Disease on Brain Magnetic Resonance Imaging: The Atherosclerosis Risk in Communities (ARIC) Study.
Chronic kidney disease (CKD) is a risk factor for cognitive decline, but evidence is limited on its etiology and morphological manifestation in the brain. We evaluated the association of estimated glomerular filtration rate (eGFR) and urinary albumin-creatinine ratio (UACR) with structural brain abnormalities visible on magnetic resonance imaging (MRI). We also assessed whether this association was altered when different filtration markers were used to estimate GFR. Cross-sectional study nested in a cohort study. 1,527 participants in the Atherosclerosis Risk in Communities (ARIC) Study. Log(UACR) and eGFR based on cystatin C, creatinine, cystatin C and creatinine in combination, or β2-microglobulin (B2M). Brain volume reduction, infarcts, microhemorrhages, white matter lesions. Multivariable linear and logistic regression models fit separately for each predictor based on a 1-IQR difference in the predictor value. Each 1-IQR lower eGFR was associated with reduced cortex volume (regression coefficient: -0.07 [95% CI, -0.12 to-0.02]), greater white matter hyperintensity volume (logarithmically transformed; regression coefficient: 0.07 [95% CI, 0.01-0.15]), and lower white matter fractional anisotropy (regression coefficient: -0.08 [95% CI, -0.17 to-0.01]). The results were similar when eGFR was estimated with different equations based on cystatin C, creatinine, a combination of cystatin C and creatinine, or B2M. Higher log(UACR) was similarly associated with these outcomes as well as brain infarcts and microhemorrhages (odds ratios per 1-IQR-fold greater UACR of 1.31 [95% CI, 1.13-1.52] and 1.30 [95% CI, 1.12-1.51], respectively). The degree to which brain volume was lower in regions usually susceptible to Alzheimer disease and LATE (limbic-predominant age-related TDP-43 [Tar DNA binding protein 43] encephalopathy) was similar to that seen in the rest of the cortex. No inference about longitudinal effects due to cross-sectional design. We found eGFR and UACR are associated with structural brain damage across different domains of etiology, and eGFR- and UACR-related brain atrophy is not selective for regions typically affected by Alzheimer disease and LATE. Hence, Alzheimer disease or LATE may not be leading contributors to neurodegeneration associated with CKD.


Characterization of the methanotrophic bacterial community present in a trichloroethylene-contaminated subsurface groundwater site.
Groundwater, contaminated with trichloroethylene (TCE) and tetrachloroethylene (PCE), was collected from 13 monitoring wells at Area M on the U.S. Department of Energy Savannah River Site near Aiken, S.C. Filtered groundwater samples were enriched with methane, leading to the isolation of 25 methanotrophic isolates. The phospholipid fatty acid profiles of all the isolates were dominated by 18:1 omega 8c (60 to 80%), a signature lipid for group II methanotrophs. Subsequent phenotypic testing showed that most of the strains were members of the genus Methylosinus and one isolate was a member of the genus Methylocystis. Most of the methanotroph isolates exhibited soluble methane monooxygenase (sMMO) activity. This was presumptively indicated by the naphthalene oxidation assay and confirmed by hybridization with a gene probe encoding the mmoB gene and by cell extract assays. TCE was degraded at various rates by most of the sMMO-producing isolates, whereas PCE was not degraded. Savannah River Area M and other groundwaters, pristine and polluted, were found to support sMMO activity when supplemented with nutrients and then inoculated with Methylosinus trichosporium OB3b. The maximal sMMO-specific activity obtained in the various groundwaters ranged from 41 to 67% compared with maximal rates obtained in copper-free nitrate mineral salts media. This study partially supports the hypothesis that stimulation of indigenous methanotrophic communities can be efficacious for removal of chlorinated aliphatic hydrocarbons from subsurface sites and that the removal can be mediated by sMMO.


Who Deserves Help? Evolutionary Psychology, Social Emotions, and Public Opinion about Welfare.
Evidence suggests that our foraging ancestors engaged in the small-scale equivalent of social insurance as an essential tool of survival and evolved a sophisticated psychology of social exchange (involving the social emotions of compassion and anger) to regulate mutual assistance. Here, we hypothesize that political support for modern welfare policies are shaped by these evolved mental programs. In particular, the compassionate motivation to share with needy nonfamily could not have evolved without defenses against opportunists inclined to take without contributing. Cognitively, such parasitic strategies can be identified by the intentional avoidance of productive effort. When detected, this pattern should trigger anger and down-regulate support for assistance. We tested predictions derived from these hypotheses in four studies in two cultures, showing that subjects' perceptions of recipients' effort to find work drive welfare opinions; that such perceptions (and not related perceptions) regulate compassion and anger (and not related emotions); that the effects of perceptions of recipients' effort on opinions about welfare are mediated by anger and compassion, independently of political ideology; and that these emotions not only influence the content of welfare opinions but also how easily they are formed.


Clinical outcomes with neoadjuvant versus adjuvant chemotherapy for triple negative breast cancer: A report from the National Cancer Database.
Triple negative breast cancer (TNBC) patients frequently receive neoadjuvant chemotherapy (NAC). Only 50% will achieve pathological complete response (pCR). In this retrospective study, we evaluated TNBC outcomes with NAC vs. AC. Patients with stages II and III TNBC treated with NAC or AC between 2010 and 2013 were identified from the National Cancer Database. Baseline characteristics were compared with χ2 and two sample t tests. Kaplan-Meier survival analyses were computed in patients treated with NAC or AC, and log-rank tests used to examine differences. Unadjusted analyses of trends in proportions over time were performed using Cochran-Armitage tests. Log-binomial models were applied to estimate relative risks of non-pCR following NAC. Of 19,151 patients, 5,621 (29.4%) received NAC, 13,530 (70.6%) received AC. NAC treated patients had worse OS compared to AC treated patients (73.4% vs. 76.8%; p<0.0001). pCR rate following NAC was 47.4%, and was associated with improved 5 year OS compared to non-pCR (86.2% vs. 62.3%; p<0.0001). In patients who received NAC, age, black race, clinical stage, diagnosis year, and Charlson-Deyo comorbidity score predicted non-pCR status. Use of NAC increased over the study period from 2010 to 2013 (27.8% - 31.2%; p = 0.0002). NAC may be inferior to AC in TNBC, likely related to the high frequency of non-pCR following NAC. It is unclear if removing the primary tumor prior to chemotherapy will have a beneficial biologic impact on therapeutic efficacy. These data should be considered hypothesis-generating as it is possible that the findings are due to selection bias, as physicians may use NAC for TNBC patients with more advanced local disease. Although, NAC still has a role in TNBC, developing biomarkers to identify patients likely to achieve pCR and benefit from NAC is an urgent need.


The current status of medical marijuana in the United States.
Medical marijuana is currently a controversial issue in medicine. There are strong pro and con opinions but relatively little scientific data on which to base medical decisions. The unfortunate scheduling of marijuana in class I has limited research and only serves to fuel the controversy. This article will review the history of laws to regulate drugs in the United States in the 20th century to provide context for the current status of medical marijuana. It will include the rationale for opposing medical marijuana laws and the problem of the Schedule I inclusion of marijuana as well as other drugs. It will examine the problems associated with smoking raw marijuana and review other routes of administration. Finally, it examines the inadvisability of medicine's promotion of smoked marijuana.


The toxic shock syndrome revisited.
The association between toxic shock syndrome and the staphylococcal toxin TSST-1 is not entirely consistent. In vitro, the elaboration of the toxin occurs only under aerobic conditions, whereas the environment of the vagina is largely anaerobic. So far, moreover, the materials of which tampons are composed seem not especially to favour the growth of the causative micro-organisms. Further, the relationship between the disease and tampon absorbability is not entirely established. Nevertheless, the effect of particular fibres cannot be discounted.


Handling Ibuprofen increases pain tolerance and decreases perceived pain intensity in a cold pressor test.
Pain contributes to health care costs, missed work and school, and lower quality of life. Extant research on psychological interventions for pain has focused primarily on developing skills that individuals can apply to manage their pain. Rather than examining internal factors that influence pain tolerance (e.g., pain management skills), the current work examines factors external to an individual that can increase pain tolerance. Specifically, the current study examined the nonconscious influence of exposure to meaningful objects on the perception of pain. Participants (N = 54) completed a cold pressor test, examined either ibuprofen or a control object, then completed another cold pressor test. In the second test, participants who previously examined ibuprofen reported experiencing less intense pain and tolerated immersion longer (relative to baseline) than those who examined the control object. Theoretical and applied implications of these findings are discussed.


Predictions based on inflammatory cytokine profiling of Egyptian COVID-19 with 2 potential therapeutic effects of certain marine-derived compounds.
A worldwide coronavirus pandemic has affected many healthcare systems in 2019 (COVID-19). Following viral activation, cytokines and chemokines are released, causing inflammation and tissue death, particularly in the lungs, resulting in severe COVID-19 symptoms such as pneumonia and ARDS. COVID-19 induces the release of several chemokines and cytokines in different organs, such as the cardiovascular system and lungs. COVID-19 and its more severe effects, such as an elevated risk of death, are more common in patients with metabolic syndrome and the elderly. Cytokine storm and COVID-19 severity may be mitigated by immunomodulation targeting NF-κB activation in conjunction with TNF- α -inhibition. In severe cases of COVID-19, inhibiting the NF-κB/TNF- α, the pathway may be employed as a therapeutic option. The study will elaborate on the Egyptian pattern for COVID-19 patients in the first part of our study. An Egyptian patient with COVID-19 inflammatory profiling will be discussed in the second part of this article using approved marine drugs selected to inhabit the significant inflammatory signals. A biomarker profiling study is currently being performed on Egyptian patients with SARS-COV-2. According to the severity of the infection, participants were divided into four groups. The First Group was non-infected with SARS-CoV-2 (Control, n = 16), the Second Group was non-intensive care patients (non-ICU, n = 16), the Third Group was intensive care patients (ICU, n = 16), and the Fourth Group was ICU with endotracheal intubation (ICU + EI, n = 16). To investigate COVID-19 inflammatory biomarkers for Egyptian patients, several inflammatory, oxidative, antioxidant, and anti-inflammatory biomarkers were measured. The following are examples of blood tests: CRP, Ferritin, D-dimer, TNF-α, IL-8, IL-6., IL-Ib, CD8, NF-κB, MDA, and total antioxidants. The results of the current study revealed many logical findings, such as the elevation of CRP, Ferritin, D-dimer, TNF- α, CD8, IL-6, IL-, NF-κB, and MDA. Where a significant increase showed in ICU group results (23.05 ± 0.30, 2.35 ± 0.86, 433.4 ± 159.3, 26.67 ± 3.51, 7.52 ± 1.48, 7.49 ± 1.04, 5.76 ± 1.31, 7.41 ± 0.73) respectively, and also ICU group results (54.75 ± 3.44, 0.65 ± 0.13, 460.2 ± 121.42, 27.43 ± 2.52, 8.63 ± 2.68, 10.65 ± 2.75, 5.93 ± 1.4, 10.64 ± 0.86) respectively, as well as ICU + EI group results (117.63 ± 11.89, 1.22 ± 0.65, 918.8 ± 159.27, 26.68 ± 2.00, 6.68 ± 1.08, 11.68 ± 6.16, 6.23 ± 0.07, 22.41 ± 1.39),respectively.The elevation in laboratory biomarkers of cytokines storm in three infected groups with remarkable increases in the ICU + EI group was due to the elevation of oxidative stress and inflammatory storm molecules, which lead to highly inflammatory responses, specifically in severe patients of COVID-19. Another approach to be used in the current study is investigating new computational drug compounds for SARS-COV-2 protective agents from the marine environment. The results revealed that (Imatinib and Indinavir) had the highest affinity toward Inflammatory molecules and COVID-19 proteins (PDB ID: -7CZ4 and 7KJR), which may be used in the future as possible COVID-19 drug candidates. The investigated inflammatory biomarkers in Egyptian COVID-19 patients showed a strong correlation between IL6, TNF-α, NF-κB, CRB, DHL, and ferritin as COVID-19 biomarkers and determined the severity of the infection. Also, the oxidative /antioxidant showed good biomarkers for infection recovery and progression of the patients.


Time-dependent descending facilitation from the rostral ventromedial medulla maintains, but does not initiate, neuropathic pain.
Although injury-induced afferent discharge declines significantly over time, experimental neuropathic pain persists unchanged for long periods. These observations suggest that processes that initiate experimental neuropathic pain may differ from those that maintain such pain. Here, the role of descending facilitation arising from developing plasticity in the rostral ventromedial medulla (RVM) in the initiation and maintenance of experimental neuropathic pain was explored. Tactile and thermal hypersensitivity were induced in rats by spinal nerve ligation (SNL). RVM lidocaine blocked SNL-induced tactile and thermal hypersensitivity on post-SNL days 6-12 but not on post-SNL day 3. Lesion of RVM cells expressing mu-opioid receptors with dermorphin-saporin did not prevent the onset of SNL-induced tactile and thermal hypersensitivity, but these signs reversed to baseline levels beginning on post-SNL day 4. Similarly, lesions of the dorsolateral funiculus (DLF) did not prevent the onset of SNL-induced tactile and thermal hypersensitivity, but these signs reversed to baseline levels beginning on post-SNL day 4. Lesions of the DLF also blocked the SNL-induced increase in spinal dynorphin content, which has been suggested to promote neuropathic pain. These data distinguish mechanisms that initiate the neuropathic state as independent of descending supraspinal influences and additional mechanism(s) that require supraspinal facilitation to maintain such pain. In addition, the data indicate that these time-dependent descending influences can underlie some of the SNL-induced plasticity at the spinal level. Such time-dependent descending influences driving associated spinal changes, such as the upregulation of dynorphin, are key elements in the maintenance, but not initiation, of neuropathic states.


Anthophyllite exposure and endemic pleural plaques in Kumamoto, Japan.
This study explored the high prevalence of pleural plaques in the town of Matsubase in Kumamoto, Japan. Small-size chest X-ray film was used for screening, and all persons with pleural plaques were confirmed by computed tomography (CT). The prevalence rate of pleural plaques in the 4 districts of Matsubase and its surrounding towns and cities were also examined. The age-adjusted mortality rate for lung cancer in this town was compared with that of its surrounding towns and cities. Pleural plaques were found in 1357 persons (724 men and 633 women) among the inhabitants who were more than 20 years of age in Matsubase between 1988 and 1993. CT scans ascertained 938 cases with pleural plaques among the 11 14 persons who participated. Thus at least 9.5% of the inhabitants over 20 years of age in this town had pleural plaques. The neighboring towns had a higher rate than the more distant towns. A large-scale open-cast asbestos mine and mill had been in operation in Matsubase between 1883 and 1970. Mineral analysis revealed anthophyllite fibers. Most of the plaques were found in persons who had never worked in the mine or mill. The high prevalence of pleural plaques in Matsubase was due to anthophyllite exposure, mainly environmental. No mesotheliomas were found, however. These findings agree with those from an earlier study from Finland.


Aerosolization Performance of Jet Nebulizers and Biopharmaceutical Aspects.
In this work, 13 jet nebulizers, some of which in different configurations, were investigated in order to identify the biopharmaceutical constraints related to the quality attributes of the medicinal products, which affect their safety, efficiency, compliance, and effectiveness. The aerosolization parameters, including the aerosol output, aerosol output rate, mass median aerodynamic diameter, and fine particle fraction, were determined according to the European Standard EN 13544-1, using sodium fluoride as a reference formulation. A comparison between the aerosol output nebulization time and the fine particle fraction displayed a correlation between the aerosol quality and the nebulization rate. Indeed, the quality of the nebulization significantly increased when the rate of aerosol emission was reduced. Moreover, the performance of the nebulizers was analyzed in terms of respirable delivered dose and respirable dose delivery rate, which characterize nebulization as the rate and amount of respirable product that could be deposited into the lungs. Depending on which of these two latter parameters was used, the nebulizers showed different performances. The differences, in terms of the rate and amount of delivered aerosol, could provide relevant information for the appropriate choice of nebulizer as a function of drug product, therapy, and patient characteristics.


A case of enterolith small bowel obstruction and jejunal diverticulosis.
We reported a case of 79-year old woman with known large bowel diverticulosis presenting with small bowel obstruction due to stone impaction - found on plain abdominal X-ray. Contrast studies demonstrated small bowel diverticulosis. At laparotomy, the gall bladder was normal with no stones and no abnormal communication with small bowel - excluding the possibility of a gallstone ileus. Analysis of the stone revealed a composition of bile pigments and calcium oxalate. This was a rare case of small bowel obstruction due to enterolith formation - made distinctive by calcification (previously unreported in the proximal small bowel).


Large-scale pattern storage and retrieval using generalized brain-state-in-a-box neural networks.
In this paper, a generalized Brain-State-in-a-Box (gBSB)-based hybrid neural network is proposed for storing and retrieving pattern sequences. The hybrid network consists of autoassociative and heteroassociative parts. Then, a large-scale image storage and retrieval neural system is constructed using the gBSB-based hybrid neural network and the pattern decomposition concept. The notion of the deadbeat stability is employed to describe the stability property of the vertices of the hypercube to which the trajectories of the gBSB neural system are constrained. Extensive simulations of large scale pattern and image storing and retrieval are presented to illustrate the results obtained.


Androgen inhibits, while oestrogen enhances, restraint-induced activation of neuropeptide neurones in the paraventricular nucleus of the hypothalamus.
The hormonal response to stress is enhanced by oestrogen but inhibited by androgens. To determine underlying changes in activity of neuropeptide neurones in the paraventricular nucleus of the hypothalamus (PVN), we examined the effect of oestrogen and androgen treatment on restraint-induced c-fos mRNA, corticotropin-releasing hormone (CRH) heteronuclear RNA, and arginine vasopressin hnRNA expression in the PVN. Male rats were gonadectomized and injected with oestradiol benzoate (EB) or dihydrotestosterone propionate (DHTP; s.c., daily for 4 days). Rats were stressed by restraint for 10 min or 30 min before killing. Other rats were stressed for 30 min and then returned to their home cage for 20 min before killing. Corticosterone and adrenocorticotropic hormone responses to restraint stress were significantly greater in EB-treated rats and lower in DHTP-treated rats at the 30-min timepoint compared to controls. c-fos mRNA increases following stress were augmented by EB but inhibited by DHTP. CRH hnRNA expression increased significantly in the PVN in response to restraint stress, and this increase was augmented by EB treatment, but decreased by DHTP treatment. Vasopressin hnRNA expression was also increased in response to stress, and this increase was attenuated by DHTP. These findings indicate that gonadal hormones influence the reactivity of the hypothalamic-pituitary adrenal axis to stress.


Comparison of patellar versus hamstring tendon autografts in arthroscopic anterior cruciate ligament reconstruction: A 6-month follow-up of a randomized clinical trial.
The purpose of this study was to compare the outcomes of anterior cruciate ligament (ACL) reconstruction using the patellar versus hamstring tendon (HT) autograft. In this randomized clinical trial, fifty patients undergoing arthroscopic ACL reconstruction were randomized into two equal groups: Those treated with either autogenous patellar tendon grafts (PT group) or HT group grafts. All patients were reviewed immediately after surgery, at 6 and 12 weeks after surgery, and then at 6 months using the International Knee Documentation Committee evaluation form. Infection, severity of pain (visual analog scale), duration of rehabilitation, and clinical and magnetic resonance imaging (MRI) findings were assessed at the 6-month follow-up. Positive pivot shift and Lachman test were considered clinical signs and symptoms of treatment failure. In addition, the absence of the ACL or transverse ACL rather than the posterior oblique ligament is an MRI finding that indicates treatment failure. Comparing changes in pain and range of motion (ROM) in patients first and 6 months after therapy show that pain had been relief significantly (P < 0.001) and ROM dramatically changes (P < 0.001). The average rehabilitation period in the PT group was 13.2 ± 2.08 weeks whereas in the HT group, it was 9.28 ± 2.26 weeks. A significant difference was seen between the two groups in terms of the rehabilitation period (P < 0.001). No significant difference was found in the normal ROM between the groups (P = 0.32). When the pain severity was considered, a significant difference was found between the PT group and the HT group (P < 0.001). The HT group patients had less knee pain than did the PT group patients. No significant difference in infection rates was seen between two groups (P = 0.66). Considering the better outcomes of HT reconstructions for the two parameters of pain severity and rehabilitation period, we consider HTs to be the ideal graft choice for ACL reconstructions.


Renal osteodystrophy. Radiological diagnosis compared with a new radioimmunoassay method of parathyroid hormone.
The parathyroid hormone concentration in the blood measured by the 'two-site radioimmunoassay', which is specific for the biologically active parathyroid molecule (parathyrine), was compared with the radiological manifestations of secondary hyperparathyroidism (HPT) on magnification radiographs of the phalanges in patients undergoing maintenance hemodialysis and in patients with renal allografts. Sensitivity of radiology for the diagnosis of HPT proved to be high (88%), whereas specificity was low (30%). Statistical analysis showed that there was a good correlation between the parathyrine levels and the intensity of radiological changes in the phalanges in patients with renal allografts (coefficient of Krueger-Spearman = 0.65). In patients undergoing hemodialysis the correlation between laboratory parameters and radiological changes was poor.


Deep gray matter T2 hypointensity is present in patients with clinically isolated syndromes suggestive of multiple sclerosis.
Gray matter (GM) magnetic resonance imaging (MRI) T2 hypointensity, a putative marker of iron deposition, is a frequent finding in patients with clinically definite (CD) multiple sclerosis (MS). The objective of this study was to assess: (a) how early deep GM T2 hypointensity occurs in MS, by studying patients with clinically isolated syndromes (CIS) suggestive of MS, and (b) whether they contribute to predict subsequent evolution to CDMS. Dual-echo scans using two different acquisition protocols were acquired from 47 CIS patients and 13 healthy controls (HC). Normalized T2-intensity of the basal ganglia and thalamus was quantified. Patients were assessed clinically at the time of MRI acquisition and after three years. During the observation period, 18 patients (38%) evolved to CDMS. At the baseline, only the GM T2-intensity of the left caudate nucleus was significantly reduced in CIS patients in comparison with the HC (p = 0.04). At the baseline, the T2 intensity of the left caudate nucleus was significantly lower (p = 0.01) in CIS patients with disease dissemination in space (DIS), but not in those without DIS, compared to the HC. The baseline T2 lesion volume, but not GM T2 hypointensity, was associated with evolution to CDMS (hazard ratio = 1.60, 95% confidence interval (CI) = 1.05-2.42; p = 0.02). In CIS patients, deep GM is not spared, suggesting that iron-related changes and neurodegeneration occurs early. The magnitude of such damage is only minor and not associated with an increased risk of evolution to CDMS.


A Feature Selection and Classification Algorithm Based on Randomized Extraction of Model Populations.
We here introduce a novel classification approach adopted from the nonlinear model identification framework, which jointly addresses the feature selection (FS) and classifier design tasks. The classifier is constructed as a polynomial expansion of the original features and a selection process is applied to find the relevant model terms. The selection method progressively refines a probability distribution defined on the model structure space, by extracting sample models from the current distribution and using the aggregate information obtained from the evaluation of the population of models to reinforce the probability of extracting the most important terms. To reduce the initial search space, distance correlation filtering is optionally applied as a preprocessing technique. The proposed method is compared to other well-known FS and classification methods on standard benchmark problems. Besides the favorable properties of the method regarding classification accuracy, the obtained models have a simple structure, easily amenable to interpretation and analysis.


Therapeutic options in locally defined or advanced prostate cancer.
Critical discussion of the available therapeutic options in locally defined or advanced prostate cancer (PCa) in the absence of standards or guidelines. Medline-based survey of the pertinent literature. The therapeutic options are presented stage-by-stage. Intracapsular PCa (T1a-T2bN0M0) is preferably managed by radical prostatectomy; 'insignificant' cancers may be treated expectantly. The outcome of irradiation is not as predictable as radical surgery. The following points regarding treatment of locally advanced PCa are still under debate: neoadjuvant treatment is probably not as efficient as believed; technical refinements of radical prostatectomy may be helpful; in some instances, the value of adjuvant treatment is undecided. The efficacy of irradiation in conjunction with androgen deprivation probably equals the efficacy of surgery. No consensus can be found in the presence of positive lymph nodes, although single institutions achieve remarkable results with surgery plus androgen deprivation. Prognostic factors beside the clinical stages and grades should be incorporated in the treatment decision, and quality-of-life measurement should gain more importance when the treatment outcome is assessed.


A microwell array platform to print and measure biomolecules produced by single cells.
Here we describe a combined method to monitor the secretion of molecules produced by single cells, followed by a method to isolate the individual cells that produced these molecules. The method is based on a self-sorting microwell chip that is connected to an activated membrane that collects the produced molecules. The produced molecules are printed by diffusion in small spots onto the membrane. The location of the printed spots can be correlated to the microwell number and the cell that produced these molecules. To demonstrate the method, we used the EpCAM antibody producing hybridoma cell line VU1D9 and a genetically engineered CHO cell-line producing Her2. VU1D9 cells produced 4.6 ± 5.6 pg (mean ± SD) of EpCAM antibody per 24 h and CHO cells 6.5 ± 8.2 pg per 24 h of Herceptin antibody.


Increased risk of genetic and epigenetic instability in human embryonic stem cells associated with specific culture conditions.
The self-renewal and differentiation capacities of human pluripotent stem cells (hPSCs) make them a promising source of material for cell transplantation therapy, drug development, and studies of cellular differentiation and development. However, the large numbers of cells necessary for many of these applications require extensive expansion of hPSC cultures, a process that has been associated with genetic and epigenetic alterations. We have performed a combinatorial study on both hESCs and hiPSCs to compare the effects of enzymatic vs. mechanical passaging, and feeder-free vs. mouse embryonic fibroblast feeder substrate, on the genetic and epigenetic stability and the phenotypic characteristics of hPSCs. In extensive experiments involving over 100 continuous passages, we observed that both enzymatic passaging and feeder-free culture were associated with genetic instability, higher rates of cell proliferation, and persistence of OCT4/POU5F1-positive cells in teratomas, with enzymatic passaging having the stronger effect. In all combinations of culture conditions except for mechanical passaging on feeder layers, we noted recurrent deletions in the genomic region containing the tumor suppressor gene TP53, which was associated with decreased mRNA expression of TP53, as well as alterations in the expression of several downstream genes consistent with a decrease in the activity of the TP53 pathway. Among the hESC cultures, we also observed culture-associated variations in global gene expression and DNA methylation. The effects of enzymatic passaging and feeder-free conditions were also observed in hiPSC cultures. Our results highlight the need for careful assessment of the effects of culture conditions on cells intended for clinical therapies.


Magnesium for the Management of Chronic Noncancer Pain in Adults: Protocol for a Systematic Review.
Chronic pain is a highly prevalent and complex health problem that is associated with a severe symptom burden, as well as substantial economic and social impact. Many patients with chronic pain still suffer from unrelieved or undertreated pain due to the incomplete efficacy and dose-limiting adverse effects of current therapies. Long-term and high-dose opioid use has considerably increased in the past 20 years despite limited evidence supporting its effectiveness in several chronic pain conditions, and serious concerns have emerged regarding adverse effects and potential misuse. Until recently, the steady increase in opioid prescribing rates has been associated with rising opioid-related mortality and other serious problems, emphasizing the need for better nonopioid therapies. Emerging evidence supports the safe use of magnesium in controlling chronic pain, but its overall efficacy and safety is still unclear. This paper aims to assess the efficacy and safety of magnesium compared with a placebo for the treatment of chronic noncancer pain. We will conduct a detailed search on Cochrane Central Register of Controlled Trials, MEDLINE, and EMBASE from their inception until the date the searches are run to identify relevant randomized controlled trials. The reference lists of retrieved studies as well as Web-based trial registries will also be searched. We will include randomized double-blind trials comparing magnesium (at any dose, frequency, or route of administration) with placebo using participant-reported pain assessment. Two reviewers will independently evaluate studies for eligibility, extract data, and assess trial quality and potential bias. Risk of bias will be assessed using criteria outlined in the Cochrane Handbook for Systematic Review of Interventions. Primary outcomes for this review will include any validated measure of pain intensity or pain relief. Dichotomous data will be used to calculate the risk ratio and number needed to treat or harm. The quality of evidence will be assessed using the Grading of Recommendations Assessment, Development and Evaluation approach. This protocol is grant-funded and has undergone a peer-review process through the Queen's University Department of Anesthesiology and Perioperative Medicine Vandewater Endowed Studentship. This project is also supported, in part, by the Chronic Pain Network of the Canadian Institutes of Health Research Strategy for Patient-Oriented Research. The electronic database search strategies are currently being developed and modified. The entire review is expected to be completed by January 1, 2019. The completion of this review is expected to identify available high-quality evidence describing the efficacy and safety of magnesium for the treatment of chronic noncancer pain. PRR1-10.2196/11654.


Impact of thrombosis on pulmonary endothelial injury and repair following sepsis.
The prevailing morbidity and mortality in sepsis are largely due to multiple organ dysfunction (MOD), most commonly lung injury, as well as renal and cardiac dysfunction. Despite recent advances in defining many aspects of the pathogenesis of sepsis-related MOD, including acute respiratory distress syndrome (ARDS), there are currently no effective pharmacological or cell-based treatments for the disease. Human and animal studies have shown that pulmonary thrombosis is common in sepsis-induced ARDS, and preclinical studies have shown that anticoagulation may improve outcome following sepsis challenge. The potential beneficial effect of anticoagulation on outcome is unconvincing in clinical studies, however, and these discrepancies may arise from the multiple and sometimes opposing actions of thrombosis on the pulmonary endothelium following sepsis. It has been suggested, for example, that mild pulmonary thrombosis prevents escape of bacterial infection into the circulation, while severe thrombosis causes hypoxia and results in pulmonary endothelial damage. Evidence from both human and animal studies has demonstrated the key role of microvascular leakage in determining the outcome of sepsis. In this review, we describe thrombosis-dependent mechanisms that regulate pulmonary endothelial injury and repair following sepsis, including activation of the coagulation cascade by tissue factor and stimulation of vascular repair by hypoxia-inducible factors. Targeting such mechanisms through anticoagulant, anti-inflammatory, and reparative methods may represent a novel approach for the treatment of septic patients.


Analgesic effect of perioperative escitalopram in high pain catastrophizing patients after total knee arthroplasty: a randomized, double-blind, placebo-controlled trial.
Sufficient pain treatment remains a challenge after total knee arthroplasty (TKA), especially in high pain catastrophizing patients. Serotonergic signaling may be involved in pain processing, but the effect of selective serotonin reuptake inhibitors on well-defined postoperative pain has not previously been investigated. The authors hypothesized that perioperative escitalopram would reduce pain after TKA in high pain catastrophizing patients. A total of 120 pain catastrophizing patients (selected using the pain catastrophizing scale as preoperative screening tool) scheduled for TKA were randomized in a double-blind manner to either 10 mg escitalopram or placebo daily from preanesthesia to postoperative day 6 in addition to a standardized analgesic regime. The primary outcome was pain upon ambulation 24 h after surgery. Secondary outcomes were overall pain during well-defined mobilizations and at rest from 2 to 48 h and from days 2 to 6, morphine equivalents, anxiety, depression, and side effects. Pain upon ambulation (mean [95% CI]) 24 h after surgery in the escitalopram versus placebo group was 58 (53 to 64) versus 64 (58 to 69), the mean difference being -5 (-13 to 3), P = 0.20. Overall pain upon ambulation and at rest from days 2 to 6 was lower in the escitalopram versus placebo group, as was depression score at day 6 (all P ≤ 0.01 in analyses uncorrected for multiple tests). Side effects were nonsignificant except for reduced tendency to sweat and prolonged sleep in the escitalopram group. No other between-group differences were observed. Escitalopram did not reduce pain upon ambulation 24 h after TKA in high pain catastrophizing patients. Future studies on optimal timing, dose, and duration of selective serotonin reuptake inhibitor treatment might be warranted.


Reaction of Candida albicans cells of different morphology index with monoclonal antibodies specific for the hyphal form.
Two monoclonal antibodies (MAbs), 3D9 with reported specificity for Candida albicans hyphae, and 3B7 with reported specificity for morphological forms of C. albicans found in vivo, were tested by indirect immunofluorescence with C. albicans cells that were grown in 12 different environments (four different culture media incubated at various temperatures) and whose cellular morphology was estimated in terms of morphology index (Mi). Both MAbs reacted strongly with cells with Mi greater than 3.0, i.e., with pseudohyphal and hyphal forms, but in Eagle's medium at 26 degrees C and in a modified Sabouraud's broth medium at 30 degrees C, some reactivity was also found with cells of lower Mi (i.e., yeast forms). Therefore, it was concluded that the hyphal phenotype and the epitopes reactive with the MAbs were co-expressed but that the epitopes could also be expressed independently of the hyphal phenotype. The results confirm the propensity of C. albicans for variation of its surface antigenic composition.


Intracellular Delivery of Proteins via Fusion Peptides in Intact Plants.
In current plant biotechnology, the introduction of exogenous DNA encoding desired traits is the most common approach used to modify plants. However, general plant transformation methods can cause random integration of exogenous DNA into the plant genome. To avoid these events, alternative methods, such as a direct protein delivery system, are needed to modify the plant. Although there have been reports of the delivery of proteins into cultured plant cells, there are currently no methods for the direct delivery of proteins into intact plants, owing to their hierarchical structures. Here, we demonstrate the efficient fusion-peptide-based delivery of proteins into intact Arabidopsis thaliana. Bovine serum albumin (BSA, 66 kDa) was selected as a model protein to optimize conditions for delivery into the cytosol. The general applicability of our method to large protein cargo was also demonstrated by the delivery of alcohol dehydrogenase (ADH, 150 kDa) into the cytosol. The compatibility of the fusion peptide system with the delivery of proteins to specific cellular organelles was also demonstrated using the fluorescent protein Citrine (27 kDa) conjugated to either a nuclear localization signal (NLS) or a peroxisomal targeting signal (PTS). In conclusion, our designed fusion peptide system can deliver proteins with a wide range of molecular weights (27 to 150 kDa) into the cells of intact A. thaliana without interfering with the organelle-targeting peptide conjugated to the protein. We expect that this efficient protein delivery system will be a powerful tool in plant biotechnology.


HIV-mediated phosphatidylinositol 3-kinase/serine-threonine kinase activation in APCs leads to programmed death-1 ligand upregulation and suppression of HIV-specific CD8 T cells.
Recent evidence demonstrates that HIV-1 infection leads to the attenuation of cellular immune responses, which has been correlated with the increased expression of programmed death (PD)-1 on virus-specific CD8(+) T cells. PD-1 is induced upon T cell activation, and its prolonged expression facilitates CD8(+) T cell inhibitory signals when bound to its B7 family ligands, PD-ligand (L)1/2, which are expressed on APCs. Importantly, early reports demonstrated that blockade of the PD-1/PD-L interaction by Abs may help to counter the development of immune exhaustion driven by HIV viral persistence. To better understand the regulation of the PD-1 pathway during HIV infection, we examined the ability of the virus to induce PD-L expression on macrophages and dendritic cells. We found a direct relationship between the infection of APCs and the expression of PD-L1 in which virus-mediated upregulation induced a state of nonresponsiveness in uninfected HIV-specific T cells. Furthermore, this exhaustion phenotype was revitalized by the blockade of PD-L1, after which T cells regained their capacity for proliferation and the secretion of proinflammatory cytokines IFN-γ, IL-2, and IL-12 upon restimulation. In addition, we identify a critical role for the PI3K/serine-threonine kinase signaling pathway in PD-L1 upregulation of APCs by HIV, because inhibition of these intracellular signal transducer enzymes significantly reduced PD-L1 induction by infection. These data identify a novel mechanism by which HIV exploits the immunosuppressive PD-1 pathway and suggest a new role for virus-infected cells in the local corruption of immune responses required for viral suppression.


Improved efficiency of genotype-dependent regeneration from protoplasts of important potato cultivars.
The regeneration of protoplasts from potato (Solanum tuberosum L.) cvs. Desiree and King Edward has been significantly improved. Different shoot culture media were required for the release of viable protoplasts from cvs. Maris Piper and Desiree, and the response of protoplasts to different culture conditions depended upon the cultivar genotype of the protoplast source. Using protoplast isolation media containing 6mM CaCl2 improved protoplast viability and culture in enriched media lead to the reproducible and relatively efficient recovery of colonies from protoplasts of these cultivars. Over 70% of protoplast-derived calli from King Edward and Desiree regenerated shoots. Many shoots were grown to mature plants in soil. This is the first report of the regeneration of mature Desiree plants from protoplasts.


Association of Positive Family Relationships With Mental Health Trajectories From Adolescence to Midlife.
National longitudinal studies that examine the linkages between early family experiences and sex-specific development of depression across the life course are lacking despite the urgent need for interventions in family settings to prevent adult depression. To examine whether positive adolescent family relationships are associated with reduced depressive symptoms among women and men as they enter midlife. This study analyzed data from the National Longitudinal Study of Adolescent to Adult Health, which used a multistage, stratified school-based design to select a prospective cohort of 20 745 adolescents in grades 7 to 12 from January 3, 1994, to December 26, 1995 (wave 1). Respondents were followed up during 4 additional waves from April 14 to September 9, 1996 (wave 2); April 2, 2001, to May 9, 2002 (wave 3); April 3, 2007, to February 1, 2009 (wave 4); and March 3, 2016, to May 8, 2017 (sample 1, wave 5), when the cohort was aged 32 to 42 years. The study sample of 8952 male adolescents and 9233 female adolescents that were analyzed was a US national representation of all population subgroups by sex, race/ethnicity, socioeconomic status, and geography. Adolescent family cohesion and low parent-child conflict. Levels of depressive symptoms (Center for Epidemiologic Studies-Depression Scale [CES-D]) from ages 12 to 42 years were used to estimate propensity score-weighted growth curve models to assess sex differences in trajectories of depression by levels of positive adolescent family relationships. A total of 18 185 individuals (mean [SD] age at wave 1, 15.42 [0.12] years; 9233 [50.8%] female) participated in the study. Females and males who experienced positive adolescent family relationships had significantly lower levels of depressive symptoms from early adolescence to midlife than did those who experienced less positive adolescent family relationships. For example, depressive symptoms were lower among those with high levels of family cohesion compared with those with low cohesion between 12 (1.26 lower CES-D score; 95% CI, 1.10-1.42) and 40 (0.78 lower CES-D score; 95% CI, 0.50-1.06) years of age among females and between 12 (0.72 lower CES-D score; 95% CI, 0.57-0.86) and 37 (0.21 lower CES-D score; 95% CI, 0.00-0.41) years of age among males. The reduction in depressive symptoms associated with positive adolescent family relationships was greater for females than males during the adolescent and early adulthood years (ie, early 20s) (eg, low-high cohesion difference in mean CES-D score, -1.26 [95% CI, -1.42 to -1.10] for females and -0.72 [95% CI, -0.86 to -0.57] for males at 12 years of age; low-high cohesion difference in mean CES-D score, -0.61 [95% CI, -0.69 to -0.53] for females and -0.40 [95% CI, -0.48 to -0.31] for males at 20 years of age), after which females and males benefited equally from positive adolescent relationships throughout young adulthood to midlife. The findings suggest that positive adolescent family relationships are associated with better mental health among females and males from early adolescence to midlife. Interventions in early family life to foster healthy mental development throughout the life course appear to be important.


Therapeutic considerations in the podiatric care of ulcerations.
In this excellent article, Dr. McCarthy presents some of the most current concepts in the treatment of lower extremity ulcerations. This common clinical finding is frequently frustrating and difficult to treat effectively. It also requires a high degree of suspicion as to the primary underlying cause and many hours of careful treatment and follow-up to prevent serious secondary complications, such as infection and gangrene. The emphasis on therapeutic approaches lends much useful information in our fight with this condition and its sequelae.


Effectiveness of exercise-referral schemes to promote physical activity in adults: systematic review.
Despite the health benefits of physical activity, most adults do not take the recommended amount of exercise. To assess whether exercise-referral schemes are effective in improving exercise participation in sedentary adults. Systematic review. Studies were identified by searching MEDLINE, CINAHL, EMBASE, AMED, PsycINFO, SPORTDiscus, The Cochrane Library and SIGLE until March 2007. Randomised controlled trials (RCTs), observational studies, process evaluations and qualitative studies of exercise-referral schemes, defined as referral by a primary care clinician to a programme that encouraged physical activity or exercise were included. RCT results were combined in a meta-analysis where there was sufficient homogeneity. Eighteen studies were included in the review. These comprised six RCTs, one non-randomised controlled study, four observational studies, six process evaluations and one qualitative study. In addition, two of the RCTs and two of the process evaluations incorporated a qualitative component. Results from five RCTs were combined in a meta-analysis. There was a statistically significant increase in the numbers of participants doing moderate exercise with a combined relative risk of 1.20 (95% confidence intervals = 1.06 to 1.35). This means that 17 sedentary adults would need to be referred for one to become moderately active. This small effect may be at least partly due to poor rates of uptake and adherence to the exercise schemes. Exercise-referral schemes have a small effect on increasing physical activity in sedentary people. The key challenge, if future exercise-referral schemes are to be commissioned by the NHS, is to increase uptake and improve adherence by addressing the barriers described in these studies.


Time course of vascular reactivity to contracting and relaxing agents after endothelial denudation by balloon angioplasty in rat carotid artery.
Endothelial injuries induced by different stimuli lead to proliferation of intimal vascular smooth muscle cells with formation of neointima. In this functional study, we evaluated the reactivity to contracting and vasorelaxing agents in Wistar rat carotid artery at different times (1, 7, 14, 21 and 28 days) after endothelial denudation with angioplastic balloon technique. Injured (IC) and uninjured carotid artery rings (UC) were placed in an isolated organ bath for isometric force displacement. IC collected at 1, 7, 14, 21 and 28 days showed a reduction in contraction to phenylephrine (0.3 microM), angiotensin II (0.1 microM), U46619 (0.1 microM), KCl (60 mM) and A23187 (1microM) at any experimental time compared to rings obtained from UC. The evaluation of endothelial-derived relaxing or hyperpolarizing factor (EDRF or EDHF), induced by acetylcholine (0.001-1 microM) in presence of indomethacin (10 microM) or indomethacin and Nomega-nitro-L-arginine methyl ester hydrochloride (L-NAME) (10 and 100 microM, respectively), was carried out at 14, 21 and 28 days. The EDRF-induced relaxation was significantly (P < 0.0001) reduced at 14 days and it improved through out the observation time, indeed at 28 days it was indistinguishable from UC relaxation curve. In contrast, the EDHF-induced relaxation was significantly (P < 0.0001) reduced at all experimental time. A significant reduction in nitric oxide-induced relaxation, sodium nitroprusside (0.001-10 microM), was observed at 7, 14 and 21 days, but not at 28 days. The relaxation induced by diazoxide (3-300 microM), an opener of KATP channels, was significantly reduced only at 7 days but not at 14, 21 and 28 days. Western blot analysis of myosin heavy chain revealed that up to 28 days the re-differentiation (maturity state) of smooth muscle cells was not yet reached. In conclusion, our data showed that hyporeactivity to contracting and relaxing agents in endothelial denuded carotid of rats could be linked to a multifactorial condition in which reduction of receptors and alterations in post-receptor transductions in neointima may produce modification of protein expression and/or variation in ion flux where calcium could have a pivotal role.


Response of plasma LH and FSH to synthetic LH-RH in children at various pubertal stages.
Fifty normal boys and 47 normal girls, aged 5 to 18 years, underwent a standard luteinizing hormone-releasing hormone (LH-RH) test (50 mug/sq m by rapid intravenous injection), and the plasma levels of LH and follicle-stimulating hormone (FSH) were determined. The results were analyzed separately according to the pubertal stages for each sex. A wide range of distribution of the individual measurements of the plasma gonadotropins throughout the LH-RH tests was found, but the mean values of the basal and peak levels showed a definite pattern for each sex at the different pubertal stages. Of particular interest was the sharp rise in basal plasma FSH level and its marked response to LH-RH in girls at the onset of puberty. The girls at this stage had the highest basal and peak FSH levels obtained at any pubertal stage in both sexes. It is concluded that the establishment of norms of LH and FSH response to a standard dose of LH-RH will be useful in evaluating normal and abnormal pubertal states in both sexes.


Circular PVT1 regulates cell proliferation and invasion via miR-149-5p/FOXM1 axis in ovarian cancer.
Long non-coding RNA plasmacytoma variant translocation 1 (PVT1) is a dysregulated gene in malignancy and is associated with oncogenesis. In this study, we found PVT1 RNA was an ovarian specific expressing gene, and overexpressed in multiple cancer types, including ovarian cancer (OV). Higher expression levels of PVT1 are related to shorter survival time in OV patients, especially in patients with advanced stage and grade. Recent studies indicated circular PVT1 also had an important role in cancer progression, whose roles in OV remain unclear. Knockdown of circular PVT1 significantly suppressed OV cell proliferation, migration and invasion. Bioinformatics analysis demonstrated that circular PVT1 was involved in regulating angiogenesis, osteoblast differentiation, regulation of cell growth, type B pancreatic cell proliferation, negative regulation of apoptotic process, phospholipid homeostasis, regulation of neurogenesis, definitive hemopoiesis, cell migration, regulation of glucose metabolic process, central nervous system development and type 2 immune response. Our data showed miR-149-5p targeted FOXM1, which was regulated by circular PVT1. Forkhead Box M1 (FOXM1) expression in ovarian cancer exhibited high level when compared with normal tissues, and had relation with relatively poor survival. FOXM1 promoted cell viability and reduced FOXM1 could rescue circular influence of circular PVT1-caused carcinoma induction. In conclusion, circular PVT1 increased FOXM1 level via binding to miR-149-5p and thus affected ovarian cancer cell viability and migration.


Investigating Dynamic High-Order Interactions in Physiological Networks through Predictive Information Decomposition.
We present an approach to assess redundant and synergistic interactions in network systems via the information-theoretic analysis of multivariate physiological processes. The approach sets up a strategy to decompose the information shared between the present states of a group of random processes and their own past states into unique contributions arising from the past of subgroups of processes and redundant and synergistic contributions arising from the dynamic interaction among the subgroups. The method is illustrated in a theoretical example of linearly interacting Gaussian processes, showing that redundancy and synergy are related mostly to unidirectional coupling and to bidirectional coupling with internal dynamics. It is then applied to the network of short-term heart period, arterial pressure and respiratory variability probed in healthy subjects, showing that redundancy and synergy prevail respectively in cardiorespiratory interactions and in cardiovascular interactions in the resting state, and that postural stress increases the predictive information and the redundancy of physiological interactions.


[Characteristics of lipid composition, lipid peroxidation intensity and ATPase activity of lymphocytes in children with acute respiratory viral infections].
Content of total lipids, intensity of lipid peroxidation and activity of ATPases were studied in lymphocytes of children with acute respiratory-viral infection (ARVI). Decrease in the content of total lipids correlated with lipid peroxidation in ARVI. At the same time, a decrease in the activity of ATPases occurred due to a decrease in the content of total lipids and alteration in the phospholipid composition of lymphocyte membranes. At the period of convalescence the patterns studied tended to normalization only in children with non-complicated forms of ARVI. Active and slowly-developed lipid peroxidation in lymphocytes of various children groups suggest dissimilar alterations in the structure and composition of the cellular membranes, which increases our knowledge on pathogenesis of ARVI.


Spectroscopic study of the transmembrane domain of a rhodopsin-phosphodiesterase fusion protein from a unicellular eukaryote.
The choanoflagellate Salpingoeca rosetta contains a chimeric rhodopsin protein composed of an N-terminal rhodopsin (Rh) domain and a C-terminal cyclic nucleotide phosphodiesterase (PDE) domain. The Rh-PDE enzyme light-dependently decreases the concentrations of cyclic nucleotides such as cGMP and cAMP. Photoexcitation of purified full-length Rh-PDE yields an "M" intermediate with a deprotonated Schiff base, and its recovery is much faster than that of the enzyme domain. To gain structural and mechanistic insights into the Rh domain, here we expressed and purified the transmembrane domain of Rh-PDE, Rh-PDE(TMD), and analyzed it with transient absorption, light-induced difference UV-visible, and FTIR spectroscopy methods. These analyses revealed that the "K" intermediate forms within 0.005 ms and converts into the M intermediate with a time constant of 4 ms, with the latter returning to the original state within 4 s. FTIR spectroscopy revealed that all-trans to 13-cis photoisomerization occurs as the primary event during which chromophore distortion is located at the middle of the polyene chain, allowing the Schiff base to form a stronger hydrogen bond. We also noted that the peptide backbone of the α-helix becomes deformed upon M intermediate formation. Results from site-directed mutagenesis suggested that Glu-164 is protonated and that Asp-292 acts as the only Schiff base counterion in Rh-PDE. A strong reduction of enzymatic activity in a D292N variant, but not in an E164Q variant, indicated an important catalytic role of the negative charge at Asp-292. Our findings provide further mechanistic insights into rhodopsin-mediated, light-dependent regulation of second-messenger levels in eukaryotic microbes.