Medically controlled glaucoma patients show greater increase in intraocular pressure than surgically controlled patients with the water drinking test.
To evaluate whether patients who have undergone trabeculectomy with mitomycin C have a different intraocular pressure (IOP) response profile after the water drinking test (WDT) from that of patients who are medically managed with a similar baseline IOP and level of visual field (VF) damage. Prospective observational study. Thirty glaucoma patients with IOP controlled by trabeculectomy and 30 with medically treated glaucoma matched for level of VF damage and IOP at baseline (7-14 mmHg). All patients underwent the WDT, which involved drinking 1000 ml of water in 15 minutes. The IOP was measured before the WDT and subsequently at 15-minute intervals for 1 hour. The peak IOP with the WDT was compared between both groups using Tukey post hoc multiple comparison and paired t tests. Baseline IOPs were 10.4+/-2.3 mmHg in surgically treated and 11.1+/-1.8 mmHg (P = 0.07) in medically treated eyes. After the WDT, mean IOPs in the surgically and medically treated groups were 10.7+/-2.3 mmHg and 14.6+/-2.2 mmHg. Mean maximum IOPs were 11.7+/-2.6 mmHg and 17.3+/-2.7 mmHg in the surgically and medically treated groups, respectively (P<0.0001), increases of 12.5% and 56%. Ranges of IOP during the WDT were 2.2+/-1.3 mmHg and 5.6+/-1.9 mmHg in the surgically and medically treated patients (P<0.0001). Patients with advanced glaucoma who are medically controlled show greater IOP elevation and peak IOP after the WDT than eyes that have undergone trabeculectomy.


Two-dimensional maps in soft immobilized pH gradient gels: a new approach to the proteome of the Third Millennium.
Same major improvements in proteome analysis of cytosolic and membrane proteins by two-dimensional mapping are here reported. A much improved transfer of proteins from the first to the second dimensional sodium dodecyl sulfate (SDS)-gel is obtained by simply diluting the gel matrix, normally composed of 4%T polyacrylamide in all commercially available Immobiline strips down to as low as 3%T. In the analysis of total lysates of platelets, this augmented transfer has been evaluated as being 2-3 times higher than in standard 4%T gels. A second major improvement, in the case of analysis of membrane protein preparations, has been demonstrated to consist in a delipidation step in a tertiary solvent mixture composed of tri-n-butyl phosphate:acetone:methanol in a 1:12:1 ratio. By adopting this protocol, large amounts of spectrins (240-220 kDa, filamentous proteins of the red blood cell membranes) could be transferred vs. essentially none when delipidation was omitted. The present report also confirms the importance of a reduction and alkylation step of the protein sample prior to all electrophoretic steps, including focusing in the Immobiline gel, as recently reported by Herbert et al.


Influence of culture time on the expression of drug-metabolizing enzymes in primary human hepatocytes and hepatoma cell line HepG2.
Primary cultures of human hepatocytes and hepatoma cell line HepG2 are frequently used to evaluate the hepatic disposition of drugs and other xenobiotics. To check the variability of the expression of drug-metabolizing enzymes in these in vitro models, expression of genes coding for several cytochrome P450 isoforms and phase II enzymes was quantified during culture time by real-time RT-PCR. Gene expression was determined daily for primary hepatocytes maintained in a sandwich culture over 1 week and for HepG2, during the first 10 passages. In primary hepatocytes characteristic expression trends were observed which could be abstracted into three major classes of time curves. Genes of the first and the second class had an expression maximum around day 6 and day 4 in culture, respectively. The third class of genes had two expression peaks: at day 1 and 5 in culture. Surprisingly, also the cell line HepG2 showed significant expression changes during passages. For example, gene expression of cytochrome 1A1 varied 8-fold, that of cytochrome 2B6 30-fold, and that of NADP-quinone reductase 1 more than 200-fold within the first 10 passages. In conclusion, neither primary hepatocytes nor HepG2 cell line display a model for constant expression of drug-metabolizing enzymes.


Annual cycle of energy and time expenditure in a golden-mantled ground squirrel population.
We have analyzed seasonal shifts of energy and time allocation in a population of golden-mantled ground squirrels (Spermophilus saturatus) by directly measuring total daily energy expenditure (DEE) with an isotopic technique ("doubly labeled water"=dlw), and by estimating components of total DEE through an integration of field behavioral observations with laboratory-measured rates of energy expenditure (oxygen consumption) associated with major behavioral and physiological states. Hibernation laster about 7 1/2 months, and the 4 1/2-month activity season consisted of mating, a 28-d gestation of 3-5 young, 5 1/2 weeks of postnatal growth building to a peak in lactation just before the young emerged above ground, an additional 2-3-week period of maternal care before dispersal, and finally restoration of body mass preceding hibernation. Although the hibernation season comprised nearly two-thirds of the year, it involved only 13-17% of annual energy expenditure, leaving about 85% of energy expenditure for the active season. Ground squirrels were actually present on the surface for only about 11% of the year's time, and the foraging time required to obtain the total annual energy supply amounted to only about 2% of the year's time. The squirrels fed mainly on herbs in the early season and hypogeous fungi later; both were used extensively during peak lactation when female energy expenditure and demand were maximal. Average daily foraging time increased steadily throughout the season to a maximum of 28% of aboveground time as availability of greens diminished and fungus predominated in the diet; time availability did not limit foraging since the animals sat on average for 65% of the daily surface time of about 7 h. Timing of reproduction is apparently optimized such that peak reproductive energy demands are matched with maximal food availability and moderate thermal conditions that minimize energy demand. Despite the greater body mass of males, the greatest total DEE (measured by dlw) of any squirrels at any time of year was that of females during peak lactation. For production of young and lactation through above-ground emergence of an average litter of 2.7, females required a total energy increase of 24% above annual nonreproductive metabolism. Yearling females all bred and performed similarly to older females, yet some costs were greater because the yearlings began and ended hibernation at smaller mass, compensated by giving birth later, and finally showed a greater absolute increase in body mass over the active season than older females. Annual metabolic energy expenditure of breeding males was about 18% greater than that of females, due to greater male body mass. Yet the annual energy intake requirement for both sexes was essentially identical (about 42MJ) due to the greater reproductive export by females in the form of newborn and milk. During the mating season males showed wide-ranging exploratory behavior and social interactions, including aggression, that involved considerable locomotory energy expenditures. Although we did not directly account for the energetics of these specific reproductive behaviors, they are critical to male reproductive success and on a daily basis they probably involved much greater energy expenditure than sperm production. Some yearling males avoided these costs by foregoing testicular development, yet they allocated four times as much energy to growth as older males, thereby increasing somatic condition for the future.


Molecular basis of film formation from a soybean protein: comparison between the conformation of glycinin in aqueous solution and in films.
Fourier transform infrared spectroscopy has been used to investigate the conformational changes of glycinin. a major storage protein of soybean seeds, upon film-forming. The results show that the secondary structure of glycinin is mainly composed of a beta-sheet (48%) and unordered (49%) structures. The amide I band of glycinin in film-forming conditions, i.e. in alkaline media and in the presence of plasticizing agent, reveals the conversion of 18% of the secondary structure of the protein from the beta-sheet (6%) and random coil (12%) to the alpha-helical conformation due to the helicogenic effect of the ethylene glycol used as the plasticizing agent. Conformational changes also occur upon the film-forming process leading to the formation of intermolecular hydrogen-bonded beta-sheet structures. Results obtained from other plant families indicate that, whatever the origin and conformation of protein, formation of films leads to the appearance of intermolecular hydrogen-bonded beta-sheet structures, suggesting that this type of structure might be essential for the network formation in films. Thus, it is hypothesized that, in the film state, intermolecular hydrogen bonding between segments of beta-sheet may act as junction zones in the film network. This study reveals for the first time that there is a close relationship between the conformation of proteins and the mechanical properties of films.


A Geographical Analysis of the African COVID-19 Paradox: Putting the Poverty-as-a-Vaccine Hypothesis to the Test.
The poverty-as-a-vaccine hypothesis came to light following the wide circulation of the controversial British Broadcasting Corporation (BBC) World Service post on the internet and social media. It was a theoretical response to what this paper has termed as "the African COVID-19 paradox" or what some have characterised as the "African COVID-19 anomaly" whose thesis is though Africa is the poorest continent in the world, yet it has some of the lowest COVID-19 infection and mortality rates globally. This paradoxical profile apparently contradicts earlier and grim projections by several international bodies on the fate of Africa in this global health crisis. Given this background, we specifically tested the validity of the hypothesis from a geographic perspective within the spatial framework of Africa. Data came from secondary sources. Evidence truly points out a significant negative relationship between COVID-19 and poverty in Africa and thus statistically supports the poverty-as-a-vaccine hypothesis. However, this does not confirm that poverty confers immunity against COVID-19 but it implicitly shows there are complex factors responsible for the anomaly. The main conclusion of the paper is that poverty has no protective immunity against COVID-19 in Africa and is therefore not tenable.


Network-based approach highlighting interplay among anti-hypertensives: target coding-genes: diseases.
Elucidating the relation between the medicines: targets, targets: diseases and diseases: diseases are of fundamental significance as-is for societal benefit. Hypertension is one of the dangerous health conditions prevalent in society, is a risk factor for several other diseases if left untreated and anti-hypertensives (AHs) are the approved drugs to treat it. The goal of the study is to decipher the connection between hypertension with other health conditions, however, is challenging due to the large interactome. To fulfill the aim, the strategy involves prior clustering of the AHs into groups as per our previous method, followed by the analyzing functional association of the target coding-genes (tc-genes) and health conditions for each group. Following our recently published work where the AHs are clustered into six groups such that molecules having similar patterns come together, here, the distribution of molecular functions and the cellular components adopted by the tc-genes of each group are analyzed. The analyses indicate that kidney, heart, brain or lung related ailments are commonly associated with the tc-genes. The association of selective tc-genes to health conditions suggests a preference for certain health conditions despite many possibilities. Analyses of experimentally validated drug-drug combinations indicate the trend in successful AHs combinations. Clinically validated combinations bind different targets. Our study provides a promising methodology in a network-based approach that considers the influence of structural diversity of AHs to the functional perspective of tc-genes concerning the health conditions. The method could be extended to explore disease-disease relationships.


Plasmodium falciparum: genetic polymorphism in apical membrane antigen-1 gene from Indian isolates.
A number of stage-specific antigens have been characterized for vaccine development against Plasmodium falciparum malaria. This study presents a comprehensive analysis of the sequence polymorphism in Plasmodium falciparum apical membrane antigen-1 (PfAMA-1) in population samples from the eastern and western parts of India. This is the first study of its kind for the nearly full length PfAMA-1 gene from these regions in India. Our observations confirmed that sequence diversity of PfAMA-1 confines only to point mutations and shows 4-8% variation as compared to the prototypes. As opposed to the previous studies on PfAMA-1, our study revealed a greater degree of polymorphism in the Domain II region of PfAMA-1 protein, though signature for diversifying selection is seen throughout the gene. Our present investigation also indicates a very high degree of variation in the reported T- and B-cell epitopes of PfAMA-1. Few noteworthy and unique observations made in this study are the substitution of Cysteine residues responsible for the disulfide bond structure of the protein and the presence of premature termination after 595 amino acids in 3 of the 13 isolates under consideration. These crucial findings add new perspectives to the future of AMA-1 research and could have major implications in establishing AMA-1 as a vaccine candidate.


The interaction effects of social support and four types of bullying on sleep quality in Chinese adolescents.
The aims of this study were to examine the association of verbal, social, physical, and cyber bullying victimizations with sleep quality while taking social support as a moderator and to further examine gender and grade differences in the moderating effects of social support on bullying-associated sleep quality among Chinese adolescents. An online cross-sectional survey was conducted in a province of northwestern China. A total of 20,320 students were included in our analyses. Multivariable linear regression analyses were conducted to examine the association between bullying victimization and sleep quality as well as the moderating effects of social support on these relationships. After adjusting for confounding variables, four types of bullying victimization were significantly associated with sleep quality. Social support only moderated the relationship of verbal, physical, and social bullying with sleep quality. Moreover, these positive moderating effects were found only for girls and, in terms of grade difference, only for primary students. Some reversed moderating effects of social support were also observed in the relationship of cyber, physical bullying with sleep quality. This was a cross-sectional study, limiting the causal inference. This study revealed that bullying is a risk factor for poor sleep quality among adolescents in northwestern China. Furthermore, social support moderated the relationship between bullying and sleep quality in different ways depending on grade, gender, and type of bullying. More efforts are needed to prevent bullying and improve both school climate and students' sleep quality.


[THE FEATURES OF SECRETION AND STOMACH MOTOR PROCESSES IN PATIENTS WITH DUODENAL ULCERS AS A CAUSE FOR REDUCING BIOAVAILABILITY OF PROTON PUMP INHIBITORS].
The article contains the modern data about the role of duodenal reflux in the development of acid-dependent diseases of stomach and duodenum. There are the results of studies in which the known quantitative parameters of duodenal reflux in the duodenal ulcer patients were simulated to identify their influence on the disintegration of enterosoluble dosage forms and the bioavailability of proton pump inhibitors (in healthy volunteers, patients with gasritis, exacerbation and remission of peptic ulcer disease the pharmacokinetics of lansoprazole, intragastric pH-metry with floating capsule for teleradio-pH-metry were investigated, and in an in vitro study the dissolution tests of proton pump inhibitors with pH 3 and pH = 4 were made). The high-amplitude oscillations of intragastric pH, observed during exacerbation of duodenal ulcer, can lead to an early disintegration of enteric-coated dosage form of proton pump inhibitors in the stomach, its destruction and decreased bioavailability.


Probing DNA structure and function with a multi-wavelength fluorescence confocal laser microscope.
Three levels of organization in DNA structure in the interphase cell nucleus are assessed by confocal laser scanning microscopy: (i) the conformational state of the double helix; (ii) the distribution of eu- and heterochromatin; and (iii) the localization of replication complexes throughout S phase. Multi-parameter measurements were carried out in each optical section using two laser sources and combined stereoscopic reconstructions were used to assess the co-localization of nuclear components. DNA is highly polymorphic and can adopt a variety of different helical conformations as well as unusual structures (curved, cruciform, multi-stranded). We have assessed by laser scanning microscopy the presence of left-handed Z-DNA in polytene chromosomes of Diptera as well as the spatio-temporal distribution of Z-DNA binding proteins in whole-mount Drosophila embryos and ovaries. We have determined the 3-D distribution of replication sites relative to heterochromatin regions, nucleoli and nuclear membrane by using short pulses of BrdU incorporation in synchronized mouse and human fibroblasts. Replication sites were visualized with a monoclonal anti-BrdU antibody combined with DNA fluorescent staining and antibody labelling of nuclear lamin. The implications of dynamic DNA movement and structural rearrangement to the organization of the nucleus in domains are discussed.


Procedural characteristics of pulmonary vein isolation using the novel third-generation cryoballoon.
A novel third-generation cryoballoon (CB3) to perform pulmonary vein isolation (PVI) has recently been released, featuring a shortened distal balloon tip when compared with the second-generation (CB2), possibly allowing for enhanced intra-ablation pulmonary vein (PV) signal mapping. We aimed to investigate procedural efficacy and safety of the CB3 as compared to the CB2. We studied 472 consecutive patients who underwent CB-PVI for paroxysmal or persistent atrial fibrillation (CB3: 49 patients; CB2: 423 patients). Detailed procedural data and in-hospital complications were registered in a prospective database. Complete PVI using the CB only was achieved in 98% of patients in each group. Single-freeze PVI was observed in 84/88% (CB2/CB3, P = n.s.) of the PVs. Time-to-PVI (TPVI) was 49 ± 32 (CB2) and 45 ± 27 s (CB3) (P = n.s.). Time-to-PVI determination rate was higher in the CB3 group (89.5 vs. 82.6%, P = 0.016). Signal noise due to ice formation on mapping electrodes occurred after 70 ± 46 s using CB3 and did not interfere with TPVI determination. Exchange of the spiral mapping catheter with a guide wire was more frequently required in the CB3 group (8.2 vs. 0.7% patients, P < 0.001). Balloon dislodgement during hockey stick manoeuvres occurred in 6.1% patients of the CB3 group only (P = 0.001). Complication rates were not different between the groups. The CB3 offers a higher TPVI determination rate, facilitating dosing schemes based on TPVI, with equally high single-freeze efficacy compared with the CB2. The shortened distal tip of the CB3 requires adaptation of standard catheter manoeuvers to avoid balloon dislodgement.


Taurine and its chloramine: modulators of immunity.
Taurine is a semiessential amino acid that is not incorporated into proteins. In mammalian tissues, taurine is ubiquitous and is the most abundant free amino acid in the heart, retina, skeletal muscle, and leukocytes. Taurine reaches up to 50 mM concentration in leukocytes. Taurine has been shown to be tissue-protective in many models of oxidant-induced injury. One possibility is that taurine reacts with HOCl, produced by the myeloperoxidase (MPO) pathway, to produce the more stable but less toxic taurine chloramine (Tau-Cl). However, data from several laboratories demonstrate that Tau-Cl is a powerful regulator of the immune system. Specifically, Tau-Cl has been shown to downregulate the production of proinflammatory mediators in both rodent and human leukocytes. Recent molecular studies on the function of taurine provide evidence that taurine is a constituent of biological macromolecules. Specifically, two novel taurine-containing modified uridines have been found in both human and bovine mitrochondria. In studies on mechanism of action, Tau-Cl inhibits the activation of NFkappaB, a potent signal transducer for inflammatory cytokines, by oxidation of IkappaB alpha at methionine45. Taurine transporter knockout mice show reduced taurine, reduced fertility, and loss of vision resulting from severe retinal degeneration, which was found to be due to apoptosis. Apoptosis induced by amino chloramines is a current and important finding because oxidants derived from leukocytes play a key role in killing pathogens. The fundamental importance of taurine in adaptive and acquired immunity will be revealed using genetic manipulation.


Large, single-center, single-operator experience with transvenous lead extraction: outcomes and changing indications.
Lead extraction is increasingly necessary given the exponential growth in cardiac device implantation. Meanwhile, the tools, indications, and outcomes of this procedure continue to change. The purpose of this study was to examine contemporary indications, outcomes, and complications of transvenous lead extraction in a large series of patients at a high-volume lead extraction center. We performed a retrospective cohort study of consecutive patients undergoing lead extraction at a single, high-volume center. Patient and lead characteristics and the indications, outcomes, and need for laser assistance were analyzed. From January 2000 to March 2007, a total of 975 chronic endovascular leads were removed from 498 patients. Median implant duration was 5.7 years (range 0.5-32.7 years). Indications were infection (60.3%), mechanical lead failure (29.3%), and upgrade of device system (8.8%). Over the study period, lead malfunction decreased relative to other indications. Laser assistance for extraction was more likely with leads implanted longer than 3.4 years compared to less than 3.4 years (odds ratio 6.15, 95% confidence interval 3.35-11.28) and with implantable cardioverter-defibrillator leads compared to pacemaker leads (odds ratio 3.44, 95% confidence interval 1.84-6.43). Overall, 97.5% of the leads were completely removed. Major complications occurred in 2 (0.4%) patients. Only one patient required cardiac surgery. No deaths occurred. In a high-volume center, lead extraction has a high success rate and low complication rate. Infection was the most common indication overall. Lead failure has decreased in relative proportion. Implantable cardioverter-defibrillator leads and longer lead implant time are associated with a requirement for laser lead extraction.


Aberrant DNA methylation in non-small cell lung cancer-associated fibroblasts.
Epigenetic changes through altered DNA methylation have been implicated in critical aspects of tumor progression, and have been extensively studied in a variety of cancer types. In contrast, our current knowledge of the aberrant genomic DNA methylation in tumor-associated fibroblasts (TAFs) or other stromal cells that act as critical coconspirators of tumor progression is very scarce. To address this gap of knowledge, we conducted genome-wide DNA methylation profiling on lung TAFs and paired control fibroblasts (CFs) from non-small cell lung cancer patients using the HumanMethylation450 microarray. We found widespread DNA hypomethylation concomitant with focal gain of DNA methylation in TAFs compared to CFs. The aberrant DNA methylation landscape of TAFs had a global impact on gene expression and a selective impact on the TGF-β pathway. The latter included promoter hypermethylation-associated SMAD3 silencing, which was associated with hyperresponsiveness to exogenous TGF-β1 in terms of contractility and extracellular matrix deposition. In turn, activation of CFs with exogenous TGF-β1 partially mimicked the epigenetic alterations observed in TAFs, suggesting that TGF-β1 may be necessary but not sufficient to elicit such alterations. Moreover, integrated pathway-enrichment analyses of the DNA methylation alterations revealed that a fraction of TAFs may be bone marrow-derived fibrocytes. Finally, survival analyses using DNA methylation and gene expression datasets identified aberrant DNA methylation on the EDARADD promoter sequence as a prognostic factor in non-small cell lung cancer patients. Our findings shed light on the unique origin and molecular alterations underlying the aberrant phenotype of lung TAFs, and identify a stromal biomarker with potential clinical relevance.


Folate supplementation during methotrexate therapy for patients with psoriasis.
Methotrexate is a folate antagonist that is a well-established therapy for autoimmune and inflammatory conditions. In some patients, methotrexate is associated with significant side effects and toxicity. Folate supplementation is often used to ameliorate methotrexate-associated side effects and toxicities. We sought to demonstrate that folate supplementation during methotrexate therapy reduces both toxicity and side effects without compromising efficacy. A MEDLINE search of the search terms "methotrexate," "folic acid," "folinic acid," and "leucovorin" was performed and literature relevant to the use of folates as a supplement to methotrexate was reviewed. According to studies reviewed, the use of folate supplements in patients treated with methotrexate reduces the incidence of hepatotoxicity and gastrointestinal intolerance without impairing the efficacy of methotrexate. Both folic acid and folinic acid are equally effective; however, folic acid is more cost effective. It must be noted that there are relatively few studies that have addressed folate supplementation with the use of methotrexate for the treatment of psoriasis. After examining the available data from the literature and drawing from clinical experience, we advise folate supplementation for every patient who receives methotrexate.


The fate of 8-methoxypsoralen-photoinduced DNA interstrand crosslinks in Fanconi's anemia cells of defined genetic complementation groups.
The fate of 8-methoxypsoralen (8-MOP)-photoinduced DNA interstrand crosslinks was followed by alkaline elution in Fanconi's anemia (FA) fibroblasts belonging to complementation groups A (FA 150 and FA 402) and B (FA 145) in comparison to a normal (1 BR/3) and a heterozygote (F 311) cell line. Clonogenic cell survival to 8-MOP photoaddition was established in parallel for all cell lines. In comparison to normal cells, group A FA cells demonstrated a higher photosensitivity than group B cells (sensitivity index 2.3 and 1.5, respectively), the heterozygote cell line being only slightly more sensitive. FA cells from both groups A and B demonstrated an incision capacity of crosslinks, the kinetics and extent of which being, however, different from that of normal or heterozygote cells. The incision is slower in FA cells and, at 24 h of post-treatment incubation, the amount of crosslinks incised is clearly lower than that observed in normal cells for group A cells, whereas in group B cells incision approaches the level of normal cells. These results correlate with survival as well as with rates of DNA semi-conservative synthesis after 8-MOP photoaddition.


Responses of stream algae to grazing minnows and nutrients: a field test for interactions.
Previous studies have shown that an algivorous grazing minnow (Campostoma anomalum) is the major herbivore in Brier Creek, a hardwater stream in south central Oklahoma. In summer and autumn schools of Campostoma virtually eliminate algae from substrate surfaces in deeper areas of some pools. The pool-to-pool distributions of algae and Campostoma reported for this stream could occur if nutrient limitation permits grazing by Campostoma to "outrun" algal growth. To test this hypothesis, mesh pens were built to exclude Campostoma from substrates in each of four typical Campostoma pools. N+P+K lawn fertilizer was added daily to two of the four pools; the other two, which received no fertilizer additions and which were not visibly affected by fertilizer transported downstream from the pools enriched with nutrients, served as controls. Algae accumulated rapidly on natural substrates and on unglazed ceramic tiles in grazer-exclusion pens in pools receiving N+P+K additions and more slowly in pens in both control pools. Periphyton biomass on grazed substrates in all four pools remained low throughout the experiment. Hence, Campostoma at normal densities were able to outrun algal growth even when nutrients were added. Eleven days after the experiment started, I determined biomass, biomass-specific net primary productivity, and areal net primary productivity of periphyton on substrates exposed to all combinations of grazer (+,0) and nutrient (+,0) treatments. Grazing increased biomass-specific primary productivity, prevented accumulation of biomass, and decreased areal primary productivity of periphyton. Additions of N+P+K increased biomass-specific net primary productivity of grazed and ungrazed periphyton and markedly increased biomass of periphyton on substrates protected from Campostoma. Although food supply for Campostoma appeared to be greater with nutrient additions, condition of Campostoma in pools receiving N+P+K was not significantly different from Campostoma collected from control pools 35 days after the experiment started. I conclude that although nutrient supply limits biomass-specific primary productivity of periphyton in Brier Creek, nutrient limitation in this stream exacerbates, rather than causes, the visually conspicuous pool-to-pool complimentary distribution of algae and Campostoma: in this stream, grazing by Campostoma at natural densities can outrun periphyton growth even when nutrients are added.


Human flexor tendon participation in the in vitro repair process.
This work demonstrates that human flexor tendon segments have the same capabilities to survive culturing and to mobilize an intrinsic healing response as previously described animal models. Segments of human flexor profundus tendons from the flexor tendon sheath were cultured in vitro and analyzed at 1, 2, and 4 weeks by scanning and transmission electron microscopy. The epitenon area thickened during the first week of culture. Fibroblasts migrated over the cut surface of the collagen bundles until they encapsulated the cut end by the second week when new collagen fibrils were seen in the extracellular spaces. By the fourth week, cells in multiple layers covered the cut end of the tendon, both collagen fibrils and new mature collagen bundles were seen. End encapsulation was accomplished by fibroblasts migrating from the outer surface and the interstices of the tendon. The encapsulating cells were active in phagocytosis and collagen synthesis during the entire incubation period.


The effect of appointment rescheduling on monitoring interval and patient attendance in the glaucoma outpatient clinic.
To assess the effect of appointment rescheduling on monitoring intervals and patient attendance in the glaucoma clinic. A retrospective review of 100 consecutive patients was conducted. Patients were categorised into groups based on risk of progressive visual loss. The monitoring interval requested by the clinician was compared with the interval recommended by national guidelines. One hundred consecutive patients who had had their appointment rescheduled were also examined to assess the effect of appointment rescheduling on the actual monitoring interval. Patient non-attendance rates and attendances at the emergency department were also examined. After excluding patients with secondary glaucoma, glaucoma suspects, and those with short-term factors affecting monitoring interval, 54 patients with chronic open-angle glaucoma (COAG) were included. Forty-eight (89%) of patients had a monitoring interval requested in accordance with national guidelines (2 had longer intervals by 1-2 months and 4 had shorter time intervals). The monitoring interval was not influenced by disease severity (Kruskal-Wallis test, P=0.16), but was significantly shorter if the intraocular pressure was above target (P<0.0001) or the patient showed structural or functional progression (P<0.0001). Hospital-initiated appointment rescheduling led to significant lengthening of monitoring interval. The mean difference between intended and actual monitoring interval was 5.6 months. Eight percent (8/100) of patients with rescheduled appointments did not attend compared with 15% (39/265) without rescheduled appointments. Clinical staff are selecting appropriate monitoring intervals for patients with COAG; however, hospital-initiated rescheduling of appointments is a major challenge to appropriate follow-up.


Psychoneuroimmunology and related mechanisms in understanding health disparities in vulnerable populations.
The nervous system as well as the endocrine system maintain extensive communication with the immune system through the influence of hormones and neurotransmitters and also by way of the hardwiring of sympathetic and parasympathetic nerves to the lymphoid organs. There is now convincing evidence that the communication between these three body systems is bidirectional. This chapter will provide a succinct review of how neuroendocrine and immune functions are affected in factors that impact vulnerability, such as aging, acute infection, and central nervous system injury. Given that the relevant literature on these topics is vast, the presentation in this chapter will serve to highlight primary references that reflect state of the science in these systems of focus.


Neodymium: YAG laser iridotomies--short-term comparison with capsulotomies and long-term follow-up.
This study addresses three aspects of anterior segment Nd:YAG laser treatment--acute endothelial damage, as assessed by endothelial specular photomicrography (ESP), acute and long-term intraocular pressure (IOP) changes, and long-term iridotomy patency. The acute ESP and IOP changes in 26 eyes (21 patients) after Nd:YAG laser iridotomies were compared to 39 eyes (37 patients) after Nd:YAG laser capsulotomy. Similar endothelial damage occurred in both groups, although less damage was noted in the group of 9 eyes in which capsulotomies were undertaken in the presence of an intraocular lens. In a parallel study 53 eyes (44 patients) were followed for a mean of 83 weeks (19 months) from the time of Nd:YAG iridotomy. There were no late closures and no late rises in IOP. The level of acute IOP rise after treatment did not predict long-term IOP. We conclude that Nd:YAG iridotomy is an effective procedure in the long-term, and that both iridotomy and capsulotomy are accompanied by noteworthy acute endothelial changes and intraocular pressure rises.


NFAT and NF-κB dynamically co-regulate TCR and CAR signaling responses in human T cells.
While it has been established that the responses of T cells to antigens are combinatorially regulated by multiple signaling pathways, it remains elusive what mechanisms cells utilize to quantitatively modulate T cell responses during pathway integration. Here, we show that two key pathways in T cell signaling, calcium/nuclear factor of activated T cells (NFAT) and protein kinase C (PKC)/nuclear factor κB (NF-κB), integrate through a dynamic and combinatorial strategy to fine-tune T cell response genes. At the cis-regulatory level, the two pathways integrate through co-binding of NFAT and NF-κB to immune response genes. Pathway integration is further regulated temporally, where T cell receptor (TCR) and chimeric antigen receptor (CAR) activation signals modulate the temporal relationships between the nuclear localization dynamics of NFAT and NF-κB. Such physical and temporal integrations together contribute to distinct modes of expression modulation for genes. Thus, the temporal relationships between regulators can be modulated to affect their co-targets during immune responses, underscoring the importance of dynamic combinatorial regulation in cellular signaling.


Research Techniques Made Simple: Optical Clearing and Three-Dimensional Volumetric Imaging of Skin Biopsies.
Skin histology is traditionally carried out using two-dimensional tissue sections, which allows for rapid staining, but these sections cannot accurately represent three-dimensional structures in skin such as nerves, vasculature, hair follicles, and sebaceous glands. Although it may be ideal to image skin in a three-dimensional manner, it is technically challenging to image deep into tissue because of light scattering from collagen fibrils in the dermis and refractive index mismatch owing to the presence of differing biological materials such as cytoplasm, and lipids in the skin. Different optical clearing methods have been developed recently, making it possible to render tissues transparent using different approaches. Here, we discuss the steps involved in tissue preparation for three-dimensional volumetric imaging and provide a brief overview of the different optical clearing methods as well as different imaging modalities for three-dimensional imaging.


In-vitro cytotoxicity assessment of carbon-nanodot-conjugated Fe-aminoclay (CD-FeAC) and its bio-imaging applications.
We have investigated the cytotoxic assay of Fe-aminoclay (FeAC) nanoparticles (NPs) and simultaneous imaging in HeLa cells by photoluminescent carbon nanodots (CD) conjugation. Non-cytotoxic, photostable, and CD NPs are conjugated with cationic FeAC NPs where CD NPs play a role in bio-imaging and FeAC NPs act as a substrate for CD conjugation and help to uptake of NPs into cancer cells due to positively charged surface of FeAC NPs in physiological media. As increase of CD-FeAC NPs loading in HeLa cell in vitro, it showed slight cytotoxicity at 1000 μg/mL but no cytotoxicity for normal cells up to concentration of 1000 μg/mL confirmed by two 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and neutral red (NR) assays, with further observations by 4',6-diamidino-2-phenylindole (DAPI) stained confocal microscopy images, possessing that CD-FeAC NPs can be used as potential drug delivery platforms in cancer cells with simultaneous imaging. Graphical abstract CD conjugation with organo-building blocks of delaminated FeAC NPs.


Optimization of acquisition parameters for cortical inhomogeneous magnetization transfer (ihMT) imaging using a rapid gradient echo readout.
Imaging biomarkers with increased myelin specificity are needed to better understand the complex progression of neurological disorders. Inhomogeneous magnetization transfer (ihMT) imaging is an emergent technique that has a high degree of specificity for myelin content but suffers from low signal to-noise ratio (SNR). This study used simulations to determine optimal sequence parameters for ihMT imaging for use in high-resolution cortical mapping. MT-weighted cortical image intensity and ihMT SNR were simulated using modified Bloch equations for a range of sequence parameters. The acquisition time was limited to 4.5 min/volume. A custom MT-weighted RAGE sequence with center-out k-space encoding was used to enhance SNR at 3 T. Pulsed MT imaging was studied over a range of saturation parameters, and the impact of the turbo factor on the effective ihMT resolution was investigated. 1 mm isotropic ihMTsat maps were generated in 25 healthy adults. Greater SNR was observed for larger number of bursts consisting of 6-8 saturation pulses each, combined with a high readout turbo factor. However, that protocol suffered from a point spread function that was more than twice the nominal resolution. For high-resolution cortical imaging, we selected a protocol with a higher effective resolution at the cost of a lower SNR. We present the first group-average ihMTsat whole-brain map at 1 mm isotropic resolution. This study presents the impact of saturation and excitation parameters on ihMTsat SNR and resolution. We demonstrate the feasibility of high-resolution cortical myelin imaging using ihMTsat in less than 20 min.


Patellofemoral contact pressures exceed the compressive yield strength of UHMWPE in total knee arthroplasties.
To address mechanisms involved in wear and permanent deformation of patellofemoral components in total knee arthroplasties, a previously reported knee joint loading model and pressure-sensitive film were used to measure patellofemoral contact areas and pressures in human cadaver knee joints after implantation with six different total knee joint designs. The joints were tested at three different Q angles (physiologic, -10 degrees, and +10 degrees) and four different flexion angles (30 degrees, 60 degrees, 90 degrees, and 120 degrees). Patellofemoral contact areas at normal Q angles ranged from 0.13 to 0.68 cm2 and increased with flexion angle up to 90 degrees. These contact areas differed significantly with flexion angle but not with Q angle. Variations in contact area with type of knee system were only marginally significant (P < .04), and post hoc tests showed no significant differences between individual knee designs. Contact pressures at normal Q angle also increased with flexion angle and ranged from 10 MPa to more than 49 MPa. Contact pressures at flexion angles greater than 60 degrees were, for all systems, well in excess of the compressive yield strength of ultrahigh-molecular-weight polyethylene and at least three to four times greater than the recommended maximum compressive stress level of 10 MPa.


Is aortic wall degeneration related to bicuspid aortic valve anatomy in patients with valvular disease?
Patients with bicuspid aortic valve are at increased risk for aortic complications. A total of 115 consecutive patients with bicuspid aortic valve disease underwent surgery of the ascending aorta. We classified the cusp configuration by 3 types: fusion of left coronary and right coronary cusps (type A), fusion of right coronary and noncoronary cusps (type B), and fusion of left coronary and noncoronary cusps (type C). Histopathologic changes in the ascending aortic wall were graded (aortic wall score). We observed type A fusion in 85 patients (73.9%), type B fusion in 28 patients (24.3%), and type C fusion in 2 patients (1.8%). Patients with type A fusion were younger at operation than patients with type B fusion (51.3 +/- 15.5 years vs 58.7 +/- 7.6 years, respectively; P = .034). The mean ascending aorta diameter was 48.9 +/- 5.0 mm and 48.7 +/- 5.7 mm in type A and type B fusion groups, respectively (P = .34). The mean aortic root diameter was significantly larger in type A fusion (4.9 +/- 6.7 mm vs 32.7 +/- 2.8 mm; P < .0001). The aortic wall score was significantly higher in type A fusion than in type B fusion (P = .02). The prevalence of aortic wall histopathologic changes was significantly higher in type A fusion. Moreover, there were no statistically significant differences between type A and type B fusion in terms of prevalence of bicuspid aortic valve stenosis, regurgitation, or mixed disease. In diseased bicuspid aortic valves, there was a statistically significant association between type A valve anatomy and a more severe degree of wall degeneration in the ascending aorta and dilatation of the aortic root at younger age compared with type B valve anatomy.


Effectiveness of a comprehensive tele-empowerment programme on self-care behaviours, uncertainty and readmission of patients with heart failure: protocol for a randomised controlled trial.
Heart failure (HF) as a long-term clinical syndrome is associated with inadequate self-care behaviours, a feeling of uncertainty and frequent hospitalisation. In recent years, empowerment has evolved for improving chronic disease management. Nevertheless, there is a lack of studies investigating remote care interventions such as a tele-empowerment programme in patients with HF. Therefore, this protocol proposes a randomised controlled trial which aims to evaluate the effectiveness of a comprehensive tele-empowerment programme on self-care behaviours, uncertainty and readmission in patients with HF. The study is a double-arm and parallel-group randomised controlled trial in which a 10-week intervention, including 6 weeks of a comprehensive tele-empowerment programme and 4 weeks of follow-up, will be compared with usual care. A total of 96 eligible patients with HF will be recruited and randomly assigned to the intervention or control group. The patients in the intervention group will join virtual groups and receive the five-step tele-empowerment programme through the internet. The primary outcomes include self-care behaviours and uncertainty which will be measured with valid instruments at baseline and 10th week. The secondary outcome is the number of patients' hospital readmissions and will be assessed at the end of the study. Descriptive statistics will be used to describe variables. According to the types of variables, appropriate statistical tests including two-sample t-tests, Χ2, analysis of covariance or linear regression will be performed. In addition, standardised intervention effect sizes will be calculated for each outcome. The trial has been approved by the Research Ethics Committee of School of Nursing and Midwifery & Rehabilitation at Tehran University of Medical Sciences. In this study, written consent will be obtained from all participants. The results will be presented to representative groups and published in peer-reviewed journals. Iranian Registry of Clinical Trials (IRCT20100725004443N30).


Altered functional connectivity between cortical networks associated with inhibitory control in trauma-exposed females.
Post-traumatic stress disorder (PTSD) is associated with impaired inhibitory control and alterations in large-scale brain network connectivity. However, few studies to date have examined the construct of inhibitory control as it relates to resting-state functional connectivity (rsFC) in a population with PTSD or trauma-exposure. The present study investigated the relationship between impaired inhibitory control and rsFC within the default mode network (DMN), central executive network (CEN), and salience network (SN) in a sample of females exposed to interpersonal trauma with and without PTSD (n = 67). Participants completed a classic Color-Word Stroop task as a measure of inhibitory control and two resting-state fMRI scans. We conducted voxelwise rsFC analyses with seed regions in the DMN, CEN, and SN and voxelwise linear regression analyses to examine the relationship between inhibitory control and rsFC of these networks across the sample. Better Stroop performance was negatively associated with total self-reported PTSD symptoms. An analysis of PTSD symptom clusters indicated that better Stroop performance was also associated with re-experiencing and hyperarousal symptoms, but not avoidance PTSD symptoms. Decreased coupling between the CEN and the DMN was associated with better inhibitory control in this sample of trauma-exposed females. These findings lend support to the hypothesis that efficient switching between these networks may contribute to better performance on cognitive and attentional tasks in trauma-exposed individuals.


Who is attending? End-of-life decision making in the intensive care unit.
Traditional expectations of the single attending physician who manages a patient's care do not apply in today's intensive care units (ICUs). Although many physicians and other professionals have adapted to the complexity of multiple attendings, ICU patients and families often expect the traditional, single physician model, particularly at the time of end-of-life decision making (EOLDM). Our purpose was to examine the role of ICU attending physicians in different types of ICUs and the consequences of that role for clinicians, patients, and families in the context of EOLDM. Prospective ethnographic study in a university hospital, tertiary care center. We conducted 7 months of observations including 157 interviews in each of four adult critical care units. The term "attending physician" was understood by most patients and families to signify an individual accountable person. In practice, "the attending physician" was an ICU role, filled by multiple physicians on a rotating basis or by multiple physicians simultaneously. Clinicians noted that management of EOLDM varied in relation to these multiple and shifting attending responsibilities. The attending physician role in this practice context and in the EOLDM process created confusion for families and for some clinicians about who was making patient care decisions and with whom they should confer. Any intervention to improve the process of EOLDM in ICUs needs to reflect system changes that address clinician and patient/family confusion about EOLDM roles of the various attending physicians encountered in the ICU.


A clinical investigation of chronic pancreatitis--comparative study between alcoholic pancreatitis and non-alcoholic pancreatitis--.
An analysis of 89 cases with chronic pancreatitis revealed 56% of chronic alcoholic pancreatitis (AP). The sex ratio was 47 males to 3 females and the mean age at onset was 42.4 years in AP and 49.3 years in chronic non-alcoholic pancreatitis (NAP). The 'total amount' of alcohol consumption was correlated to the onset. In AP, the abdominal pain was apt to relapse and severe in nature, furthermore painless pancreatitis was seen in 6%. The association with diabetes or calcification (38%) were more frequently seen in AP. The calcifications in AP appeared to be smaller in size and distributed diffusely or localized in cephalic portion. A striking frequency of liver dysfunctions (39 cases) were demonstrated, however, cirrhosis was rare in AP. The P-S test dysunctions in NAP were frequently reversible in the follow up study, while even some of chronic asymptomatic alcoholics developed clinical signs of pancreatitis during the observed period and proceeded to definite AP, of whom pancreatic dysfunctions showed fluctuation and eventual progression. In other words, even pain free intervals the pancreatic inflammation in susceptible persons may proceed to ultimated destruction of the pancreas. The fatality from chronic pancreatitis was rare (12.3%), which was related diabetes mellitus.


Evaluation of phenolic profile and antioxidant capacity in gluten-free flours.
The characterization of phenolic fingerprints in common gluten-free flours is still scarce. Total phenolic and anthocyanin contents, antioxidant capacity and the entire phenolic profile were investigated for extracts from chickpea, sorghum, quinoa, black rice, lentil, amaranth, brown rice, oat and white rice flours, using soft wheat flour as a comparison. The highest phenolic content was found in black rice, followed by quinoa extracts (147.9 and 87.2mg gallic acid equivalents 100g-1, respectively). Consistently, antioxidant capacity was highest in black rice and quinoa flours (34mmol trolox equivalents 100g-1), while anthocyanins were highest in black rice flour. Data showed a high correlation between phenolic content and both reducing and scavenging activities, with Pearson's coefficient of 0.90 and 0.91, respectively. Although the entire phenolic profile was diverse and differed among flours, these represent a valuable source of health-promoting compounds, mainly belonging to flavonoids, phenolic acids and lignans.


Atrial Fibrillation Following Coronary Artery Bypass Graft: Where Do We Stand?
Atrial fibrillation (AF) is the most common atrial arrhythmia following coronary artery bypass graft (CABG). Its prevalence is 15-45% and is associated with poor long-term prognosis. Risk factors can be patient-related, intraoperative, and/or postoperative. Therapeutic and preventive strategies have been developed to curtail AF burden. Cardioversion is recommended for unstable or symptomatic patients and rate control if asymptomatic. Anticoagulation is challenging with risk of thromboembolism and bleeding. However, patients should be anticoagulated after cardioversion or if AF persists >48 h and risk factors of stroke exist. A minimum of 4 weeks is recommended but longer duration should be considered in patients at high risk of stroke irrespective of recurrence of AF.


Simultaneous quantification of 17 trace elements in blood by dynamic reaction cell inductively coupled plasma mass spectrometry (DRC-ICP-MS) equipped with a high-efficiency sample introduction system.
A quadrupole inductively coupled plasma mass spectrometer (Q-ICP-MS) equipped with a dynamic reaction cell (DRC) and coupled with a desolvating nebulization system (APEX-IR) was employed to determine 17 elements (Al, As, Ba, Cd, Co, Cr, Li, Mn, Mo, Ni, Pb, Sb, Se, Sn, Sr, V, and Zr) in blood samples. Ammonia (for Al, Cr, Mn, and V) and O2 (for As and Se) were used as reacting gases. Selection of the best flow rate of the gases and optimization of the quadrupole dynamic bandpass tuning parameter (RPq) were carried out, using digested blood diluted 1+9 with deionized water and spiked with 1 microg L(-1) of Al, Cr, Mn, V and 5 microgL(-1) of As and Se. Detection limits were determined in digested blood using the 3sigma criterion. The desolvating system allowed a sufficient sensitivity to be achieved to determine elements at levels of ng L(-1) without detriment of signal stability. The accuracy of the method was tested with the whole blood certified reference material (CRM), certified for Al, As, Cd, Co, Cr, Mn, Mo, Ni, Pb, Sb, Se, and V, and with indicative values for Ba, Li, Sn, Sr, and Zr. The addition calibration approach was chosen for analysis. In order to confirm the DRC data, samples were also analyzed by means of sector field inductively coupled plasma mass spectrometry (SF-ICP-MS), operating in medium (m/Deltam=4000) and high (m/Deltam=10,000) resolution mode and achieving a good agreement between the two techniques.


Studies of an acidic cardiotoxin isolated from the venom of Mojave rattlesnake (Crotalus scutulatus).
A major lethal protein was isolated from the venom of Mojave rattlesnake (Crotalus scutulatus) by successive purification in DEAE column chromatography and isoelectric focusing. This homogeneous and monomeric form of toxin is designated as "Mojave toxin". Unlike basic neurotoxins or cytotoxins isolated from venoms of cobras, kraits and sea snakes, the Mojave toxin is an acidic protein with an isoelectric point of 4.7. The toxin is also different from crotoxin (from Crotalus durissus terrificus) which consists of both acidic and basic components. The molecular weight determined by Sephadex G-75 column chromatography resulted in a value of about 22 000. A singel protein band with a molecular weight of about 12 000, was observed after sodium dodecyl sulfate gel electrophoresis of the reduced Mojave toxin. Isoelectric focusing gel in the presence of 8 M urea also showed a single protein band, suggesting that the toxin is composed of subunits. Unlike the neurotoxic nature of the basic proteins from the venoms of Elapidae and sea snakes (Hydrophiidae) and crotoxin, Mojave toxin is cardiotoxic rather than neurotoxic. It is very likely that venoms of all rattlesnakes from North and Central America contain Mojave toxin as the common toxin.


Stereumin A-E, sesquiterpenoids from the fungus Stereum sp. CCTCC AF 207024.
Five cadinane sesquiterpenoids, named stereumin A (1), B (2), C (3), D (4) and E (5) were isolated from the CHCl(3) extract of the culture broth of the fungal strain CCTCC AF 207024. Based on the sequences at the internal transcribed spacer (ITS) region and partial 28S rDNA, this fungus was identified as a Stereum sp. The structures of the five compounds were elucidated using spectroscopic data from 1D, 2D NMR and HRESIMS experiments, and the structures of 1 and 2 were further confirmed by single-crystal X-ray diffraction analysis. Compounds 1-5 showed nematicidal activities against the nematode Panagrellus redivivus at 400 mg l(-1). Among these five compounds, compounds 3 and 4 killed 84.4% and 94.9% of P. redivivus, respectively in 48 h.


Role of unilateral percutaneous cervical cordotomy in the treatment of neoplastic vertebral pain.
The present study is a long-term report on 69 patients undergoing unilateral percutaneous cervical cordotomy for vertebral pain due to neoplastic bone metastases. The pain was unilateral or bilateral and was characterised by a chronic and/or an incident component. Seventy-one per cent (49/69) of the patients benefitted from the operation, obtaining complete, lasting pain relief or a reduced degree of pain, amenable to control by medication with narcotic or non-narcotic drugs. There was a survival Q(50%) of 5 months (S.E. = 1.6) with no pain and a satisfactory quality of life.


Surface analysis of titanium implants.
Medical grade titanium samples were examined using X-ray photoelectron spectroscopy before and after immersion in various proteins. Additionally, an implant removed from a patient following clinical failure was examined using scanning ion and electron microscopy. The surface of the as-received samples was found to be mainly TiO2, with contaminants of H2O/OH-, calcium and nitrogen which remained after autoclaving. The immersed proteins adhered to the titanium surface, possibly via a Ca-O link. The failed clinical sample was found to be partially fibrously encapsulated with evidence of calcification. Small amounts of TiOOH were detected at the fibrous periphery, supporting the theory of Tengvall and co-workers that in vivo titanium implants are covered in a gel of this material.


[Detection of significant gender differences by odontometry as illustrated by a representative group from Lebanese (East-Mediterranean) population].
This study was conducted on a sample of the Lebanese population. Its aim was to verify differences in dental dimensions between the genders as was found by other authors in other populations. It is concluded that gender dental differences constitute an additional important, though not-decisive element, in identification of courpses.


Protein kinase inhibitor design by targeting the Asp-Phe-Gly (DFG) motif: the role of the DFG motif in the design of epidermal growth factor receptor inhibitors.
The Asp-Phe-Gly (DFG) motif plays an important role in the regulation of kinase activity. Structure-based drug design was performed to design compounds able to interact with the DFG motif; epidermal growth factor receptor (EGFR) was selected as an example. Structural insights obtained from the EGFR/2a complex suggested that an extension from the meta-position on the phenyl group (ring-5) would improve interactions with the DFG motif. Indeed, introduction of an N,N-dimethylamino tail resulted in 4b, which showed almost 50-fold improvement in inhibition compared to 2a. Structural studies confirmed this N,N-dimethylamino tail moved toward the DFG motif to form a salt bridge with the side chain of Asp831. That the interactions with the DFG motif greatly contribute to the potency of 4b is strongly evidenced by synthesizing and testing compounds 2a, 3g, and 4f: when the charge interactions are absent, the inhibitory activity decreased significantly.


Medical image compression using 3-D Hartley transform.
In this paper, 3-D discrete Hartley transform is applied for the compression of two medical modalities, namely, magnetic resonance images and X-ray angiograms and the performance results are compared with those of 3-D discrete cosine and Fourier transforms using the parameters such as PSNR and bit rate. It is shown that the 3-D discrete Hartley transform is better than the other two transforms for magnetic resonance brain images whereas for the X-ray angiograms, the 3-D discrete cosine transform is found to be superior.


Video-enhanced microscopic visualization of apoptotic cell death caused by anti-Fas antibody in living human glioma cells.
To study the morphological changes of anti-Fas antibody-mediated apoptosis in living U251-SP human glioma cells, we employed video-enhanced contrast differential interference contrast (VEC-DIC) microscopy. In our previous study, we investigated the susceptibility of human glioma cell lines to anti-Fas Immunoglobulin M (IgM) antibody. U251-SP cells express Fas antigen on their surface. The cells exposed to anti-Fas antibody underwent apoptotic cell death, as reported previously. In this study, morphological changes of apoptosis characterized by bleb formation, shrinkage of cells, and nuclear condensation were observed under VEC-DIC microscopy in U251-SP human glioma cells treated with anti-Fas antibody. These results demonstrate the usefulness of VEC-DIC microscopy to study the process of apoptotic cell death.


A thermodynamic study on the interaction between magnesium ion and human growth hormone.
A thermodynamic study on the interaction between magnesium ion and human growth hormone (hGH) was studied at 27 degrees C in NaCl solution (50 mM) using different techniques. Two techniques of ionmetry using a Mg2+selective membrane electrode and isothermal titration calorimetry were applied to obtain the binding isotherm for hGHMg2+; results obtained by both techniques were found to be in good agreement. There is a set of three identical and noninteracting binding sites for magnesium ions. The intrinsic dissociation equilibrium constant and the molar enthalpy of binding are 46 microM and -17.7 kJ/mol, respectively. Temperature scanning UV-visible spectroscopy was applied to elucidate the effect of Mg2+ binding on the protein stability, and circular dichroism (CD) spectroscopy was used to show the structural change of hGH due to the metal ion interaction. Magnesium ion binding increased the protein thermal stability by increasing the alpha-helix content as well as decreasing both beta and random coil structures. However, the secondary structural change of the protein returns to its native form, including a small change in the tertiary structure, in high concentrations of magnesium ion.


Influence of weather conditions on children's school travel mode and physical activity in 3 diverse regions of Canada.
Children who engage in active school transportation (AST) have higher levels of physical activity (PA). Climate and weather were shown to influence adults' daily travel behaviours, but their influence on children's AST and PA has been less examined. This study examined the influence of weather conditions on children's AST and overall PA. Children in grades 4 to 6 (N = 1699; age, 10.2 ± 1.0 years) were recruited in schools located in urban, suburban and rural areas, stratified by area-level socioeconomic status, in 3 different regions of Canada (Trois-Rivières, Québec; Ottawa, Ontario; Vancouver, British Columbia). Mode of school travel was self-reported and physical activity was measured using a pedometer. We used publicly available data on total precipitation and early morning temperature. AST increased with temperature only among girls. Daily precipitation was negatively associated with boys' and girls' PA while warmer temperature was associated with increased PA on weekend days. We also observed that season and region moderated the relationship between weather conditions and children's physical activity behaviours. Our results suggest that daily weather variations influence children's AST and PA to a greater extent than seasonal variations. Interventions designed to help children and families adapt to weather-related barriers to AST and PA are needed. Novelty: In Canada, weather conditions may influence children's active behaviours daily. Associations between weather conditions, choice of travel mode and physical activity vary by sex, season, and region. Weather affects children's PA differently during the week than on weekends.


The air meniscus as a radiographic finding: a review of the literature and presentation of nine unusual cases.
In addition to echinococcal cysts and mycetomas, a wide variety of disorders can occasionally produce an air meniscus sign on a chest radiograph. A proposed classification follows: I. Infections A. Lung abscess (with or without pulmonary gangrene) B. Fungus ball C. Bacterial ball D. Tuberculoma E. Blood clot in tuberculous cavity, Rasmussen aneurysm F. Echinococcal lung cyst II. Neoplastic A. Bronchogenic carcinoma B. Primary lung sarcoma C. Metastatic carcinoma, sarcoma to lung D. Bronchial adenoma E. Cystic hamartoma III. Developmental A. Bochdalek hernia (pseudocavity) IV. Traumatic A. Pulmonary hematoma V. Hemodynamic A. Congestive heart failure (with or without bullae)


Kinetics of carrier-mediated ion transport in two new types of solvent-free lipid bilayers.
In contrast with the usual glyceryl-monooleate/decane (GMO-D) bilayer lipid membranes, new membranes, formed from a mixture of GMO in squalene (GMO-S) or from a mixture of GMO in triolein (GMO-T), seem to be almost solvent free. Our results from voltage-jump relaxation studies, using these "solvent-free" membranes with the homologue carriers, nonactin, monactin, dinactin, trinactin, and tetranactin, are compared with the corresponding ones for GMO-D membranes. With all homologues, solvent-free membranes show an increase of the free carrier translocation rate, ks, by a factor of 2.5, a decrease in the dissociation rate constant of the complex, kDi, by a factor of 1.5 and no significant change in its formation rate constant, kRi. However, the principal effect of the absence of solvent in these membranes is an increase by a factor of approximately 10 of the translocation rate constant for moving the complex across the membrane, kis. This increase varies regularly from a factor of 7-15 with decreasing carrier size, and is always larger for GMO-T than for GMO-S membranes. These solvent-free effects are interpreted in terms of modifications of electrostatic and hydrophobic energy profiles in the membrane.


Obstetric cholestasis, outcome with active management: a series of 70 cases.
To determine the nature and outcome of obstetric cholestasis in a United Kingdom population. Prospective analysis of clinical outcome in women diagnosed with obstetric cholestasis that is actively managed. Antenatal population of three London hospitals between August 1999 and April 2001. Seventy women with obstetric cholestasis defined as abnormal liver function (one or more abnormality in gamma-glutamyl transpeptidase, alanine amino-transferase, aspartate amino-transferase and total bile acids) in a pregnant woman with pruritus, in the absence of other pathology. All women were interviewed weekly regarding their symptoms. All were actively managed according to a standardised protocol, which included early delivery before 38 weeks. Obstetric outcome was recorded. Seventy women of mean age 30 (6) years delivered 73 infants. The median gestation at onset of pruritus was 30 (range 4-39) weeks and at diagnosis of obstetric cholestasis was 33.7 (range 21-40.7) weeks. Asian women were more likely to be diagnosed with obstetric cholestasis. Pruritus was usually severe and generalised, and commonly worst on the palms and/or soles of the feet. There were no stillbirths or perinatal deaths. Twenty-five women required caesarean section (36%); only four (16%) were for fetal distress. Twelve women (17%) delivered before 37 weeks, of which eight (67%) were iatrogenic. Ten (14%) infants required admission to the special care baby unit of which four (40%) were ventilated. Policies of active management result in increased intervention and associated complications. This must be balanced against possible reductions in perinatal mortality.


Influence of magnesium on the secretion and action of parathyroid hormone.
The interactions of serum levels of magnesium and parathyroid hormone (PTH) have been studied in a patient with hypomagnesemia and hypocalcemia following intestinal bypass surgery for obesity. When serum magnesium was low serum PTH was not stimulated by hypocalcemia. With correction of magnesium deficiency hypocalcemia was associated with elevation of serum PTH levels. Infusion of exogenous PTH induced a clearly detectable renal response in the presence of hypomagnesemia but the response was diminished when serum magnesium was elevated. In this patient it appears that hypomagnesemia suppressed parathyroid gland activity, leaving the renal action of PTH intact.


Surface charge and zeta-potential of metabolically active and dead cyanobacteria.
Zeta potential and acid-base titrations of active, inactivated, and dead Planktothrix sp. and Synechococcus sp. cyanobacteria were performed to determine the degree to which cell surface electric potential and proton/hydroxyl adsorption are controlled by metabolism or cell membrane structure. Surface OH(-) excess from potentiometric data, showed differences in surface charge between active and dead cyanobacteria from pH 3 to 10. Average zero salt effect pH (pH(pzse)) of 5.8+/-0.1 and 6.3+/-0.1 were obtained for active Planktothrix sp. and Synechococcus sp., respectively. Similarly for dead cyanobacteria pH(pzse) values of 5.8+/-0.1 and 4.6+/-0.1 were obtained. Zeta potentials of active Planktothrix sp. and Synechococcus sp. were positive at alkaline conditions, with a maximum of +13.7+/-1.5 mV at a pH of 9.0+/-0.1 for both species. This positive potential diminished in the presence of 1 mM HCO(-)(3). The zeta potential of Planktothrix sp. and Synechococcus sp. cells was negative at alkaline pH following their exposure to NaN(3), a metabolic inhibitor. The zeta potential of dead cyanobacteria was negative for Planktothrix sp., from pH 2.5 to 10.5, at -30 to -20 mV. Dead Synechococcus sp. exposed to a pH 2.5 solution recorded negative potentials to a minimum of -30 mV at pH 8, but positive potentials were found at higher pH reaching a maximum of +10 mV at pH 9.1. Zeta potentials for dead, but non-acidified Synechococcus sp. remained negative at -30 mV from an initial pH of 5.6 to 10.5, reflecting differences in cell wall structure between these species. These results indicate that Planktothrix sp. and Synechococcus sp. may metabolically control their surface charge to electrostatically attract bicarbonate anions at alkaline pH, required for photosynthesis.


Gene expression and localization analysis of Bombyx mori bidensovirus and its putative receptor in B. mori midgut.
Bombyx mori bidensovirus (BmBDV), which causes fatal flacherie disease in the silkworm, replicates only in midgut columnar cells. The viral resistance expressed by some silkworm strains, which is characterized as non-susceptibility irrespective of the viral dose, is determined by a single gene, nsd-2. We previously identified nsd-2 by positional cloning and found that this gene encodes a putative amino acid transporter that might function as a receptor for BmBDV. In this study, we investigated the relationship between the part of the midgut expressing nsd-2 (resistance gene), +(nsd-2) (susceptibility gene) and BmBDV propagation. Quantitative RT-PCR (qRT-PCR) analysis using total RNA isolated from the anterior, middle, and posterior parts of the midgut showed that nsd-2 and +(nsd-2) were strongly expressed in the posterior part of the midgut. The expression levels of both genes were very low in the anterior and middle parts. The qRT-PCR analysis showed that the expression levels of BmBDV-derived transcripts were correlated with the levels of +(nsd-2) expression. However, BmBDV-derived transcripts were clearly detected in all parts of the midgut. These results suggest that the infectivity of BmBDV depends mainly on the expression level of +(nsd-2) in the midgut and that viral infection is supported even by very faint expression of +(nsd-2). By contrast, the expression levels of +(nsd-2) were exceedingly low or undetectable in the middle part of the midgut, indicating that BmBDV infection might occur via another mechanism, independent of +(nsd-2), in the middle part of the midgut.


Prenatal diagnosis of DHPR deficiency by direct detection of mutation.
Prenatal diagnosis was requested by a family carrying a 3 base-pair insertion in the dihydropteridine reductase (DHPR) coding region. A chorionic villus sample was obtained and fetal DNA was isolated directly from this. Diagnosis was performed by a polymerase chain reaction (PCR)-based technique, with a simple electrophoretic assay for the insertion. The fetus was found to be heterozygous for the insertion. This is the first time that prenatal diagnosis of DHPR deficiency has been performed by direct detection of the mutation.


On GABA metabolism in the gliocyte cells of the rat pineal gland.
The uptake of the inhibitory transmitter substance gamma-aminobutyric acid (GABA) into the adult rat pineal gland was studied autoradiographically using both light and electron microscopy. The sites of GABA uptake were shown to be exclusively present in the gliocyte cells of the gland following both in vitro incubation with tritiated GABA and after in vivo administration of the amino acid by intra-arterial injection. Both the pinealocyte cells and the numerous sympathetic axons in the gland were devoid of silver grains. Preliminary biochemical studies indicated that the gliocyte uptake process for GABA resembles that in the satellite glia of the sensory ganglia but differed from that in slices of the cerebral cortex. Evidence is also presented which shows the pineal gland to contain endogenous GABA and the enzymes directly associated with its in vivo metabolism, L-glutamate-1-carboxylase (EC 188.8.131.52) (GAD) and GABA-2-oxoglutarate aminotransferase (EC 188.8.131.52) (GABA-T). Furthermore, a 3-fold rise in endogenous GABA occurred in the pineal after inhibition of GABA-catabolism as would be expected if the GABA-shunt pathway was functionally active in the oxidative metabolism of the pineal gland.


Intravenous immunoglobulin (IVIG) preparations induce apoptosis in TNF-alpha-stimulated endothelial cells via a mitochondria-dependent pathway.
Endothelial cells (ECs) are a target in inflammation, and the death of EC is regulated by various factors. Although intravenous immunoglobulin (IVIG) preparations are known to be beneficial therapeutic agents for the treatment of autoimmune diseases and systemic inflammatory disorders, their mechanism of action have not yet been completely elucidated. The aim of the present study is to investigate the possible role of IVIG in EC apoptosis. We demonstrate herein that IVIG induced the apoptosis of human umbilical vein ECs (HUVECs) prestimulated by TNF-alpha in vitro, but not in unstimulated HUVECs, in a dose- and time-dependent manner, using a proportion of cells with hypodiploid DNA, DNA ladder formation and morphological changes. Anti-Fas MoAbs had no effect on the IVIG-induced apoptosis in the TNF-alpha-stimulated HUVECs. IVIG decreased the intracellular expression of anti-apoptotic proteins of the Bcl-2 family (A1 and Bcl-XL) while IVIG increased the intracellular expression of pro-apoptotic proteins (Bax and Bcl-XS) in the TNF-alpha-stimulated HUVECs. Furthermore, IVIG increased the intracellular production of reactive oxygen species and decreased the mitochondrial membrane potential (Delta(psi)m). Caspase-inhibitors inhibited the IVIG-induced apoptosis of the TNF-alpha-stimulated HUVECs. The present results show a novel action in which IVIG can induce the apoptosis of TNF-alpha-stimulated HUVECs through a mitochondrial apoptotic signalling pathway. These observations suggest that the clinical use of IVIG preparations may thereby regulate the cell death of activated ECs in inflammation.


Efficacy and safety of WBRT+EGFR-TKI versus WBRT only in the treatment of NSCLC patients with brain metastasis: An updated meta-analysis.
To investigate the efficacy and safety of whole brain radiotherapy (WBRT) combined with epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) versus WBRT only in the treatment of brain metastasis in non-small cell lung cancer (NSCLC) patients by pooling open published data. Prospective clinical studies relevant to WBRT+EGFR-TKI versus WBRT only in the treatment of NSCLC brain metastasis were electronically searched in the Pubmed, EMbase, Cochrane, Wangfang, CNKI and Google scholar databases. The treatment response, 1-year survival and treatment-associated toxicity were pooled and expressed by odds ratio (OR) under a fixed or random effect model. The publication bias was evaluated by Begg's funnel plot and Egger's line regression test. Eighteen prospective clinical studies were included in the study. The combined results indicated that the objective response rate (ORR) in the WBRT+TKI group was superior to WBRT only with a statistical difference (OR = 2.67, 95% CI: 2.10-3.38, p < 0.05) under a fixed effect model. Ten studies reported the 1-year survival rate between the WBRT+TKI and WBRT only groups. The combined results showed that 1-year survival rate in the WBRT+TKI group was higher than that of the WBRT only group with a statistical difference (OR = 2.70, 95% CI: 1.95-3.74, p < 0.05). For treatment-associated toxicity, the combined data indicated that the treatment-related rash in the WBRT+TKI group was significantly higher than that of the WBRT only group with a statistical difference (OR = 2.72, 95% CI: 1.53-4.84, p < 0.05). However, the incidence of nausea/vomiting (OR = 0.84, 95% CI: 0.60-1.17, p > 0.05), diarrhea (OR = 1.31, 95% CI: 0.83-2.07, p > 0.05), fatigue (OR = 1.40, 95% CI: 0.70-2.81, p > 0.05) and myelosuppression (OR = 0.86, 95% CI: 0.56-1.32, p > 0.05) were not statistically different between the two groups. Based on the current publications, WBRT+EGFR-TKI can improve the treatment response and 1-year survival rate but not increase the toxicity except for rash compared to WBRT alone in the treatment of brain metastasis in NSCLC patients.


Nacre-mimetic cerium-doped nano-hydroxyapatite/chitosan layered composite scaffolds regulate bone regeneration via OPG/RANKL signaling pathway.
Autogenous bone grafting has long been considered the gold standard for treating critical bone defects. However, its use is plagued by numerous drawbacks, such as limited supply, donor site morbidity, and restricted use for giant-sized defects. For this reason, there is an increasing need for effective bone substitutes to treat these defects. Mollusk nacre is a natural structure with outstanding mechanical property due to its notable "brick-and-mortar" architecture. Inspired by the nacre architecture, our team designed and fabricated a nacre-mimetic cerium-doped layered nano-hydroxyapatite/chitosan layered composite scaffold (CeHA/CS). Hydroxyapatite can provide a certain strength to the material like a brick. And as a polymer material, chitosan can slow down the force when the material is impacted, like an adhesive. As seen in natural nacre, the combination of these inorganic and organic components results in remarkable tensile strength and fracture toughness. Cerium ions have been demonstrated exceptional anti-osteoclastogenesis capabilities. Our scaffold featured a distinct layered HA/CS composite structure with intervals ranging from 50 to 200 μm, which provided a conducive environment for human bone marrow mesenchymal stem cell (hBMSC) adhesion and proliferation, allowing for in situ growth of newly formed bone tissue. In vitro, Western-blot and qPCR analyses showed that the CeHA/CS layered composite scaffolds significantly promoted the osteogenic process by upregulating the expressions of osteogenic-related genes such as RUNX2, OCN, and COL1, while inhibiting osteoclast differentiation, as indicated by reduced TRAP-positive osteoclasts and decreased bone resorption. In vivo, calvarial defects in rats demonstrated that the layered CeHA/CS scaffolds significantly accelerated bone regeneration at the defect site, and immunofluorescence indicated a lowered RANKL/OPG ratio. Overall, our results demonstrate that CeHA/CS scaffolds offer a promising platform for bone regeneration in critical defect management, as they promote osteogenesis and inhibit osteoclast activation.


The effectiveness of family-based treatment for full and partial adolescent anorexia nervosa in an independent private practice setting: Clinical outcomes.
Anorexia nervosa (AN) is a severe psychiatric illness with little evidence supporting treatment in adults. Among adolescents with AN, family-based treatment (FBT) is considered first-line outpatient approach, with a growing evidence base. However, research on FBT has stemmed from specialist services in research/public health settings. This study investigated the effectiveness of FBT in a case series of adolescent AN treated in a private practice setting. Thirty-four adolescents with full or partial AN, diagnosed according to DSM-IV criteria, participated, and were assessed at pretreatment and post-treatment. Assessments included change in % expected body weight, mood, and eating pathology. Significant weight gain was observed from pretreatment to post-treatment. 45.9% of the sample demonstrated full weight restoration and a further 43.2% achieved partial weight-based remission. Missing data precluded an examination of change in mood and ED psychopathology. Effective dissemination across different service types is important to the wider availability of evidence-based treatments. These weight restoration data lend preliminary support to the implementation of FBT in real world treatment settings. © 2016 Wiley Periodicals, Inc. (Int J Eat Disord 2016; 49:1023-1026).


Mass spectrometric quantification of the relative amounts of C6 and C3 position phosphorylated glucosyl residues in starch.
The quantification of phosphate bound to the C6 and C3 positions of glucose residues in starch has received increasing interest since the importance of starch phosphorylation for plant metabolism was discovered. The method described here is based on the observation that the isobaric compounds glucose-6-phosphate (Glc6P) and glucose-3-phosphate (Glc3P) exhibit significantly different fragmentation patterns in negative ion electrospray tandem mass spectrometry (MS/MS). A simple experiment involving collision-induced dissociation (CID) MS(2) spectra of the sample and the two reference substances Glc3P and Glc6P permitted the quantification of the relative amounts of the two compounds in monosaccharide mixtures generated by acid hydrolysis of starch. The method was tested on well-characterized potato tuber starch. The results are consistent with those obtained by NMR analysis. In contrast to NMR, however, the presented method is fast and can be performed on less than 1 mg of starch. Starch samples of other origins exhibiting a variety of phosphorylation degrees were analyzed to assess the sensitivity and robustness of the method.


Cold shock Y-box protein-1 participates in signaling circuits with auto-regulatory activities.
The cold shock protein Y-box (YB) binding-1 is an example of a highly regulated protein with pleiotropic functions. Besides activities as a transcription factor in the nucleus or regulator of translation in the cytoplasm, recent findings indicate extracellular effects and secretion via a non-classical secretion pathway. This review summarizes regulatory pathways in which YB-1 participates, all iterating auto-regulatory loops. Schematics are developed that elucidate the cold shock protein activities in (i) fine-tuning its own expression level following platelet-derived growth factor-B-, thrombin- or interferon-γ-dependent signaling, (ii) as a component of the messenger ribonucleoprotein (mRNP) complex for interleukin-2 synthesis in T-cell commitment/activation, (iii) pro-fibrogenic cell phenotypic changes mediated by transforming growth factor-β, and (iv) receptor Notch-3 cleavage and signal transduction. Emphasis is put forward on subcellular protein translocation mechanisms and underlying signaling pathways. These have mostly been analysed in cell culture systems and rarely in experimental models. In sum, YB-1 seems to fulfill a pacemaker role in diverse diseases, both inflammatory/pro-fibrogenic as well as tumorigenic. A clue towards potential intervention strategies may reside in the understanding of the outlined auto-regulatory loops and means to interfere with cycling pathways.


Biventricular Compared to Left Ventricular Impella and Norepinephrine Support in a Porcine Model of Severe Cardiogenic Shock.
Contemporary management of cardiogenic shock (CS) with vasopressors is associated with increased cardiac workload and despite the use of unloading devices such as the Impella pump, concomitant vasopressors are often necessary. Therefore, we compared if cardiac workload could be reduced and end-organ perfusion preserved with biventricular support (Bipella) compared to ImpellaCP and norepinephrine in pigs with left ventricular (LV) CS caused by left main coronary microembolization. Cardiac workload was calculated from heart rate × ventricular pressure-volume area obtained from conductance catheters placed in the LV and right ventricle (RV), whereas organ perfusion was measured from venous oxygen saturation in the pulmonary artery (SvO 2 ) and the kidney- and the cerebral vein. A cross-over design was used to access the difference after 30 minutes of ImpellaCP and norepinephrine 0.1 µg/kg/min versus Bipella for 60 minutes. Bipella treatment reduced LV workload ( p = 0.0078) without significant difference in RV workload from ImpellaCP and norepinephrine, however a decrease in SvO 2 (49[44-58] vs . 66[63-73]%, p = 0.01) and cerebral venous oxygen saturations (62[48-66] vs . 71[63-77]%, p = 0.016) was observed during Bipella compared to ImpellaCP and norepinephrine. We conclude that Bipella reduced LV workload but did not preserve end-organ perfusion compared to ImpellaCP and norepinephrine in short-term LV CS.


An effective system of nurse-led diabetes care in rural Africa.
Delivering adequate diabetes care is difficult in rural Africa because of drug and equipment shortages; as well as lack of trained medical expertise. We aimed to set up and evaluate a nurse-led protocol and education-based system in rural Kwazulu Natal in South Africa. A treatment algorithm and education system adapted from previously validated methods was used; care was devolved to primary health clinics and was delivered by two nurses. Glycaemic control was assessed by glycated haemoglobin (HbA1c), assayed off site and not available for clinical use during the study. Results A total of 284 patients were enrolled, with 197 followed for 18 months (13 died and 26% lapsed during the period). HbA1c was 11.6 +/- 4.5% (sd) at baseline, 8.7 +/- 2.3% at 6 months and 7.7 +/- 2.0% at 18 months. There was a small associated increase in weight but no increase in hypoglycaemia. Subgroup analysis showed that education alone, without drug type or dose changes, also improved control (HbA1c 10.6 +/- 4.2% baseline and 7.6 +/- 2.3% at 18 months). The service was very popular with patients, families and other health workers. We conclude that a simple protocol and education-based diabetes care system can be successfully introduced and run by nurses in rural Africa. Medium-term glycaemic improvements are excellent and the service has been very well received.


Ca2+-cardiolipin interaction in a model system. Selectivity and apparent high affinity.
The interaction of cardiolipin with Ca2+ was assessed by measuring the cardiolipin-mediated extraction of 45Ca2+ from an aqueous to an organic (methylene chloride) phase. Cardiolipin binds Ca2+ with high affinity [Kd(apparent) = 0.70 +/- 0.17 microM (S.D.)]. Cation-cardiolipin interactions are selective. Interaction of cardiolipin with Ca2+ is insensitive to Na+, but is inhibited by divalent cations with Mn2+ greater than Zn2+ greater than Mg2+. In addition La3+ and Ruthenium red are particularly potent inhibitors of Ca2+ binding by cardiolipin. Cardiolipin-mediated extraction of Ca2+ into an aqueous phase is also inhibited by phosphatidylcholine. Inhibition of Ca2+-cardiolipin interaction by phosphatidylcholine (a phospholipid known to stabilize the bilayer conformation) may implicate inverted, non-bilayer lipid structures in the binding.


Nucleotide variation in Sabin type 3 poliovirus from an Albanian infant with agammaglobulinemia and vaccine associated poliomyelitis.
Vaccine-associated paralytic poliomyelitis (VAPP) and immunodeficient long-term polio excretors constitute a significant public health burden and are a major concern for the WHO global polio eradication endgame. Poliovirus type 3 characterized as Sabin-like was isolated from a 5-month-old Albanian child with X-linked agammaglobulinemia and VAPP after oral polio vaccine administration. Diagnostic workup and treatment were performed in Italy. Poliovirus replicated in the gut for 7 months. The 5' non coding region (NCR), VP1, VP3 capsid proteins and the 3D polymerase genomic regions of sequential isolates were sequenced. Increasing accumulation of nucleotide mutations in the VP1 region was detected over time, reaching 1.0 % of genome variation with respect to the Sabin reference strain, which is the threshold that defines a vaccine-derived poliovirus (VDPV). We identified mutations in the 5'NCR and VP3 regions that are associated with reversion to neurovirulence. Despite this, all isolates were characterized as Sabin-like. Several amino acid mutations were identified in the VP1 region, probably involved in growth adaptation and viral persistence in the human gut. Intertypic recombination with Sabin type 2 polio in the 3D polymerase region, possibly associated with increased virus transmissibility, was found in all isolates. Gamma-globulin replacement therapy led to viral clearance and neurological improvement, preventing the occurrence of persistent immunodeficiency-related VDPV. This is the first case of VAPP in an immunodeficient child detected in Albania through the Acute Flaccid Paralysis surveillance system and the first investigated case of vaccine associated poliomyelitis in Italy since the introduction of an all-Salk schedule in 2002. We discuss over the biological and clinical implications in the context of the Global Polio Eradication Program and emphasize on the importance of the Acute Flaccid Paralysis surveillance.


Effect of residential proximity to the lignite-fired power plant on depression, sleep quality, and morning salivary cortisol in the elderly.
Exposures to air pollution of by-products emission from the lignite-fired power plant elevated risks of carcinogenic effects, endocrine disruptors, central nervous system structural and functional changes. Residence in the proximity of the lignite-fired power plant appeared to have more chance to have higher risks of health problems. This study aimed to assess associations of residential proximity to the lignite-fired power plant on depression, sleep quality, and morning salivary cortisol among the elderly. The distance of residential proximity to the power plant was categorized into three groups (units in kilometer): <10 km, 10-15 km, and >15 km. The coefficients of log (morning salivary cortisol) was -0.320 (95%CI: -0.460, -0.179; p-value < 0.001) for those living <10 km compared to those living >15 km. Coefficients of sleep quality score were 1.350 (95%CI: 0.265, 2.436; p-value = 0.015) for those living <10 km compared to those living >15 km. Residential proximity to the lignite-fired power plant was not associated with depression. Our study concluded that living within 10 km to the lignite-fired power plant was related to negative health outcomes among the elderly. Policymakers need to reconsider the distance of the buffer zone to the power plant.


K+ channels and T-cell synapses: the molecular background for efficient immunomodulation is shaping up.
Diverse K(+) channels exhibit differential expression and function in human lymphocytes. Voltage-gated and Ca(2+)-activated K(+) channels have a significant impact on both T-cell activation and the overall immune response. A recent study on the plasma membrane organization of Kv1.3 K(+) channels in human T cells provides new insights into the immunoregulatory functions of K(+) channels, which might result in new and improved immune suppression protocols targeted to ion channels and/or T-cell signaling and accessory molecules.


Metabolic rewiring in melanoma.
Oncogene-driven metabolic rewiring is an adaptation to low nutrient and oxygen conditions in the tumor microenvironment that enables cancer cells of diverse origin to hyperproliferate. Aerobic glycolysis and enhanced reliance on glutamine utilization are prime examples of such rewiring. However, tissue of origin as well as specific genetic and epigenetic changes determines gene expression profiles underlying these metabolic alterations in specific cancers. In melanoma, activation of the mitogen-activated protein kinase (MAPK) pathway driven by mutant BRAF or NRAS is a primary cause of malignant transformation. Activity of the MAPK pathway, as well as other factors, such as HIF1α, Myc and MITF, are among those that control the balance between non-oxidative and oxidative branches of central carbon metabolism. Here, we discuss the nature of metabolic alterations that underlie melanoma development and affect its response to therapy.


Dehydrocostus lactone inhibits Candida albicans growth and biofilm formation.
Candida albicans infections are threatening public health but there are only several antifungal drugs available. This study was to assess the effects of dehydrocostus lactone (DL) on the Candida albicans growth and biofilms Microdilution assays revealed that DL inhibits a panel of standard Candida species, including C. albicans, as well as 9 C. albicans clinical isolates. The morphological transition of C. albicans in RPMI-1640 medium and the adhesion to polystyrene surfaces can also be decreased by DL treatment, as evidenced by microscopic, metabolic activity and colony forming unit (CFU) counting assays. The XTT assay and microscopy inspection demonstrated that DL can inhibit the biofilms of C. albicans. Confocal microscopy following propidium iodide (PI) staining and DCFH-DA staining after DL treatment revealed that DL can increase the membrane permeability and intracellular reactive oxygen species (ROS) production. N-acetyl-cysteine could mitigate the inhibitory effects of DL on growth, morphological transition and biofilm formation, further confirming that ROS production induced by DL contributes to its antifungal and antibiofilm effects. This study showed that DL demonstrated antifungal and antibiofilm activity against C. albicans. The antifungal mechanisms may involve membrane damage and ROS overproduction. This study shows the potential of DL to fight Candida infections.


Alteration of alpha-spectrin ubiquitination after hemorrhagic shock.
RBC deformability after trauma and hemorrhagic shock (T/HS) leads to the microcirculatory dysfunction and clinical manifestations of organ failure. However, the cellular mechanism of this phenomenon remains unknown. Spectrins are important for the shape and physical properties of erythrocytes, such as deformability and resistance to mechanical stress. Previous studies have shown that erythrocyte alpha-spectrin is ubiquitinated. Studies of sickled cells and aged erythrocytes, 2 conditions known to have decreased RBC deformability, have shown decreased alpha-spectrin ubiquitination, which may contribute to the inability of these cells to change shape. It was hypothesized that decreased alpha-spectrin ubiquitination could participate in the mechanism(s) whereby T/HS erythrocytes become less deformable. The level of alpha-spectrin ubiquitination in erythrocytes isolated from T/HS rats was determined and compared with erythrocytes from control sham-shocked (T/SS) animals. After T/SS (n = 4) or T/HS (n = 7), alpha- and beta-spectrin subunits were isolated using a low ionic-strength buffer at 37 degrees C for 30 minutes. The relative amount of ubiquitinated alpha-spectrin was evaluated by Western blotting using a monoclonal antibody to ubiquitin. The relative level of alpha-spectrin ubiquitination (normalized to total alpha-spectrin in the same preparation) was found to be significantly decreased after T/HS (.319 +/- .03) compared with T/SS control erythrocytes (.485 +/- .06, P < .05). To evaluate the content and relative amounts of the other membrane proteins, the profiles of T/HS and T/SS erythrocytes were compared by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. This did not reveal any significant quantitative differences between T/SS and T/HS spectrin or other membrane proteins. The finding of decreased alpha-spectrin ubiquitination after T/HS suggests that this mechanism could contribute to increased rigidity of the erythrocyte membrane.


[Significance of the ankle joint pump for returning venous blood. Report of experiences using a mechanical treatment method].
We report on new medicomechanic equipment designed for the improvement of the venous reflux in the lower extremity. It may be employed as postoperative prophylaxis against thrombosis and in the treatment of secondary manifestations based on venous insufficiency, as well as with pathological changes caused by ankylosis of the ankle.