Overexpression of arogenate dehydratase reveals an upstream point of metabolic control in phenylalanine biosynthesis.
Out of the three aromatic amino acids, the highest flux in plants is directed towards phenylalanine, which is utilized to synthesize proteins and thousands of phenolic metabolites contributing to plant fitness. Phenylalanine is produced predominantly in plastids via the shikimate pathway and subsequent arogenate pathway, both of which are subject to complex transcriptional and post-transcriptional regulation. Previously, it was shown that allosteric feedback inhibition of arogenate dehydratase (ADT), which catalyzes the final step of the arogenate pathway, restricts flux through phenylalanine biosynthesis. Here, we show that in petunia (Petunia hybrida) flowers, which typically produce high phenylalanine levels, ADT regulation is relaxed, but not eliminated. Moderate expression of a feedback-insensitive ADT increased flux towards phenylalanine, while high overexpression paradoxically reduced phenylalanine formation. This reduction could be partially, but not fully, recovered by bypassing other known metabolic flux control points in the aromatic amino acid network. Using comparative transcriptomics, reverse genetics, and metabolic flux analysis, we discovered that transcriptional regulation of the d-ribulose-5-phosphate 3-epimerase gene in the pentose phosphate pathway controls flux into the shikimate pathway. Taken together, our findings reveal that regulation within and upstream of the shikimate pathway shares control over phenylalanine biosynthesis in the plant cell.


Andrographolide Inhibits Angiogenesis by Inhibiting the Mir-21-5p/TIMP3 Signaling Pathway.
Angiogenesis provides nutrients and oxygen to promote tumor growth and affords a channel that facilitates tumor cell entry into the circulation. Andrographolide (Andro) possess anti-tumor activity; however, its direct effect on angiogenesis still needs to be clarified. In this study, our experiments revealed that Andro significantly inhibited vascular growth in chick embryo chorioallantoic membrane (CAM) and yolk sac membrane (YSM) models. Meanwhile, tumor angiogenesis was also suppressed by Andro. Additionally, we found that cell proliferation, migration and tube formation of vascular endothelial cells was inhibited by Andro treatment in vitro. The effect was primarily mediated through inhibition of miR-21-5p expression and further targeting of TIMP3. This work provides evidence that Andro directly inhibits angiogenesis and might be an effective anti-angiogenic therapeutic drug for cancer treatment.


The Influence of Practising Physical Activity on the Prevention and Treatment of Gestational Diabetes: A Systematic Review.
During pregnancy, maintaining an inadequate lifestyle (bad eating habits, stress, consumption of toxic substances, etc.) generates complications such as pre-eclampsia, overweight, lumbar pain and Gestational Diabetes. This review was carried out with the objective to determine the influence of practising physical activity on the prevention and treatment of Gestational Diabetes; and to evaluate the efficacy of interventions based on therapeutic exercise in the management of such diseases. A systematic review of the publications of the last five years in Medline, Pubmed, Web of Science and Cinahl databases was conducting with the terms Exercise and Gestational Diabetes. 25 articles were found, 9 were observational studies that analysed physical activity habits through questionnaires, and other 16 applied an intervention. With respect to the frequency of the sessions, most of these studies applied their interventions three times per week, with all of them obtaining positive results. Regarding intensity, all the studies that referred to it concluded that it must be at least moderate, highlighting that, at all times, the limitations of pregnant women must be taken into account. Both for the prevention and treatment of this disease, the physical activity must be performed for a minimum of three times per week, at least in moderate intensity, and must be based on aerobic, resistance and strength exercises.


IL-23 receptor signaling licenses group 3-like innate lymphoid cells to restrict a live-attenuated oral Chlamydia vaccine in the gut.
An IFNγ-susceptible mutant of Chlamydia muridarum is attenuated in pathogenicity in the genital tract and was recently licensed as an intracellular Oral vaccine vector or intrOv. Oral delivery of intrOv induces transmucosal protection in the genital tract, but intrOv itself is cleared from the gut (without shedding any infectious particles externally) by IFNγ from group 3-like innate lymphoid cells (ILC3s). We further characterized the intrOv interactions with ILC3s in the current study, since the interactions may impact both the safety and efficacy of intrOv as an oral Chlamydia vaccine. Intracolonic inoculation with intrOv induced IFNγ that in return inhibited intrOv. The intrOv-IFNγ interactions were dependent on RORγt, a signature transcriptional factor of ILC3s. Consistently, the transfer of oral intrOv-induced ILC3s from RORγt-GFP reporter mice to IFNγ-deficient mice rescued the inhibition of intrOv. Thus, IFNγ produced by intrOv-induced ILC3s is likely responsible for inhibiting intrOv, which is further supported by the observation that oral intrOv did induce significant levels of IFNγ-producing LC3s (IFNγ+ILC3s). Interestingly, IL-23 receptor knockout (IL-23R-/-) mice no longer inhibited intrOv, which was accompanied by reduced colonic IFNγ. Transfer of oral intrOv-induced ILC3s rescued the IL-23R-/- mice to inhibit intrOv, validating the dependence of ILC3s on IL-23R signaling for inhibiting intrOv. Clearly, intrOv induces intestinal IFNγ+ILC3s for its own inhibition in the gut, which is facilitated by IL-23R signaling. These findings have provided a mechanism for ensuring the safety of intrOv as an oral Chlamydia vaccine and a platform for investigating how oral intrOv induces transmucosal protection in the genital tract.


Sex differences in use of interventional cardiology persist after risk adjustment.
Studies from several countries have documented gender disparities in the management of coronary artery disease. Whether such gender disparities are seen in Italy and, if so, whether they can be explained by factors such as age and severity of illness were investigated. 77 974 Piedmontese patients, admitted between 1999 and 2002, with a primary diagnosis of myocardial infarction (ICD 410), angina (ICD 413), chronic ischaemia (ICD 414) and chest pain (ICD 786.5) were studied. The number of men and women undergoing surgical treatment was extracted and the male-female odds ratios calculated. Several risk factors and a risk adjustment technique (APR-DRG) were used to control for possible confounders. Backward stepwise multiple logistic regression was used to adjust for significant covariates. Crude analysis demonstrated that gender is a discriminating factor in the probability of surgery (OR 2.11, 95% CI 2.04 to 2.19), with similar findings among those with each main diagnosis. The odds ratios decreased after adjustment for age, co-morbidity and disease severity but remained significant. Men and women admitted to hospitals in a region of northern Italy with a diagnosis of cardiovascular disease are treated differently and this cannot be explained by age or severity of disease.


Use of dietary interventions for functional gastrointestinal disorders.
The role of food in the development of symptoms experienced within functional gastrointestinal disorders (FGIDs) is well recognised. This review aims to describe the evidence base for dietary interventions in the different functional esophageal, duodenal and bowel disorders. Randomised controlled trials are lacking for many of the FGIDs, with the exception of irritable bowel syndrome (IBS). Restricting rapidly fermentable, short-chain carbohydrates (FODMAPs) provides an evidence based dietary approach for the management of symptoms of IBS. Recent evidence shows the upper GI motility response varies between carbohydrates, which gives promise for the potential application of the low FODMAP diet in upper GI disorders. In addition to fine-tuning our FODMAP understanding, other observational data and smaller sized studies create an exciting and optimistic future for dietary management of all FGIDs.


Theoretical vibrational sum-frequency generation spectroscopy of water near lipid and surfactant monolayer interfaces. II. Two-dimensional spectra.
The structural stability and function of biomolecules is strongly influenced by the dynamics and hydrogen bonding of interfacial water. Understanding and characterizing the dynamics of these water molecules require a surface-sensitive technique such as two-dimensional vibrational sum-frequency generation (2DSFG) spectroscopy. We have combined theoretical 2DSFG calculations with molecular dynamics simulations in order to investigate the dynamics of water near different lipid and surfactant monolayer surfaces. We show that 2DSFG can distinguish the dynamics of interfacial water as a function of the lipid charge and headgroup chemistry. The dynamics of water is slow compared to the bulk near water-zwitterionic and water-anionic interfaces due to conformational constraints on interfacial water imposed by strong phosphate-water hydrogen bonding. The dynamics of water is somewhat faster near water-cationic lipid interfaces as no such constraint is present. Using hydrogen bonding and rotational correlation functions, we characterize the dynamics of water as a function of the distance from the interface between water and zwitterionic lipids. We find that there is a transition from bulk-like to interface-like dynamics approximately 7 Å away from a zwitterionic phosphatidylcholine monolayer surface.


Evanescent-Vacuum-Enhanced Photon-Exciton Coupling and Fluorescence Collection.
An evanescent optical mode existing in various nanophotonic structures always acts as a cavity mode rather than an electromagnetic vacuum in the study of cavity quantum electrodynamics (CQED). Here we show that taking the evanescent mode as an electromagnetic vacuum in which the nanocavity is located is possible through the optical mode design. The proposed evanescent vacuum enables us to enhance both the reversible photon-exciton interaction and fluorescence collection. By embedding the custom-designed plasmon nanocavity into the evanescent vacuum provided by a metallic or dielectric nanowire, the photon-exciton coupling coefficient can achieve 4.2 times that in vacuum due to the exponential decay of the evanescent wave, and spontaneously emitted photons with Rabi splitting can be guided by an evanescent wave with a collection efficiency of 47% at most. Electromagnetic vacuum engineering at subwavelength scale holds promise for controlling the light-matter interaction in quantum optics, CQED, and on-chip quantum information.


THEORY OF MAST REPRODUCTION IN PLANTS: STORAGE-SIZE DEPENDENT STRATEGY.
The amount of available resource can be considered to restrict mast seeding behavior in plants, regardless of the cause of masting itself. The reproductive strategy of a plant that has stopped growing, but continues to maintain a constant size, is based on the allocation of assimilated resources between reproduction and storage. If both plant death and population growth rates are constant, the plant strategy is dependent only on its own storage size. Conditions for the evolution of mast reproduction were analyzed under both constant and varying environments, from the view point of storage-size dependent strategy, which is influenced by three parameters: the advantage coefficient of mast seeding, β; the cost coefficient of delayed seeding, γ; and the assimilation rate, P. The model indicated that: (1) mast seeding evolves only with a greater than linear increase in reproductive success with effort (β > 1) under both constant and varying environments; (2) in mast seeding, a critical storage size, Sc *, occurs, above which plants utilize all storage for reproduction; (3) in a constant environment, Sc * increases with increasing β and P and/or decreasing γ; (4) concomitant with (3), an intermasting period, τ, also increases with increasing β and decreasing 7, but is independent of P; (5) in a varying environment, Sc * and the average of τ increase with increasing variance of the assimilation rate, P; and (6) concomitant with (5), the intermasting period has a certain probability distribution. In addition, reproductive synchrony within the population and the relationship between mast seeding and species diversity are discussed.


Brief Adaptation to Astigmatism Reduces Meridional Anisotropy in Contrast Sensitivity.
To investigate the effect of visual adaptation to orientation-dependent optical blur on meridional contrast sensitivity function in artificially imposed astigmatism. The study adopted a top-up adapt-test paradigm. During the blur adaptation process, the 18 non-astigmatic young adult participants were briefly presented with natural scene images (first trial, 10 minutes; subsequent trials, 6 seconds). Contrast sensitivities for horizontal and vertical gratings at spatial frequencies ranging from 1 to 8 cycles per degree (cpd) were measured immediately before and after adaptation to +3.00 diopters cylinder (DC) with-the-rule or against-the-rule astigmatism. Meridional anisotropy was measured to quantify the contrast sensitivity difference between the two grating orientations. Adapting to astigmatic blur enhanced contrast sensitivity at the blurred power meridian but reduced contrast sensitivity at the least affected axis meridian. In with-the-rule conditions, contrast sensitivity for horizontal gratings was significantly increased, whereas that for vertical gratings was significantly decreased. Similarly, in against-the-rule conditions, contrast sensitivity for vertical gratings was significantly increased, whereas that for horizontal gratings was significantly decreased. These two factors together resulted in a substantial systematic reduction, averaging 34%, in meridional anisotropy of contrast sensitivity across the spatial frequency spectrum. Astigmatism adaptation occurs in natural scene viewing. Brief exposure to astigmatic blur altered contrast sensitivity in the opposite direction at the two principal meridians, indicating that the mature visual system possesses functional plasticity to recalibrate the response characteristics of orientationally tuned cortical filters and thus promote substantial reductions of meridional anisotropy in astigmatic vision, to some extent counterbalancing the elongated oval shape of astigmatic blur.


Drug analysis at the 1988 Olympic Winter Games in Calgary.
A comprehensive drug testing program was carried out during the 16 days of the 1988 Olympic Winter Games in Calgary, Canada. State-of-the-art technology was applied, involving high-resolution gas chromatography, high-performance liquid chromatography, gas chromatography/mass spectrometry, fluorescence polarization immunoassay, and radioimmunoassay. Samples from selected athletes were screened for five drug classes: stimulants, narcotic analgesics, anabolic steroids, beta-blockers, and diuretics. In addition, samples were also screened for local anesthetics, corticosteroids, beta-human choriogonadotropin, and cannabinoids. During the 16-day event, 428 urine samples were processed and 3090 screening procedures performed. We describe the methods for analysis at the Olympic Games and present the results.


Application Experience and Patient Feedback Analysis of 3D Printed AFO with Different Materials: A Random Crossover Study.
This study is aimed at analyzing the application experience and feedback of the patients with poststroke ankle dorsiflexion disorders for 3D printed AFO with three different materials. 15 patients were randomly divided into three groups; 3D printed AFO with 3 different materials (PA2200, Somos NeXt, and PA12) was used to each group, according to the crossover study design, in order to ask the three groups of patients to use three different materials of 3D printed AFO. Assessment was taken by the end of each test round. Through statistical processing, the patient feedback data of the three groups of materials of 3D printed AFO were obtained. In the material comfort assessment of the AFO, Somos NeXt was compared with PA2200, and the p value was <0.05; in the item of surface smoothness of the AFO, Somos NeXt was compared with PA2200, and the p value was <0.01; at the same time, PA12 was compared with PA2200, and the p value was <0.05. The 3 different materials of 3D printing AFO bring different experience, and we also have sufficient reason to believe that there will be differences in the auxiliary effect of this on patients, which leads the patient's selection too. The material Somos NeXt is much popular and has certain clinical advantages.


Prenatal exposure to substance of abuse: a worldwide problem.
Substance abuse during pregnancy is an important public health issue affecting the mother and the growing infant. Preterm labor, miscarriage, abruption and postpartum hemorrhage are obstetric complications which have been associated with women who are dependent on abused substances. Moreover, women are also at an increased risk of medical problems such as poor nutrition, anemia, urinary tract infections as well as sexually transmitted infections, hepatitis, HIV and problems related to infection. Intrauterine growth restriction, prematurity, stillbirth, neonatal abstinence syndrome, and Sudden Infant Death Syndrome represent only some of fetal effects. Later on, during childhood, it has been shown that in utero exposure to substances of abuse is associated with increased rates of respiratory infections, asthma, ear and sinus infections. Moreover, these children are more irritable, have difficulty focusing their attention, and have more behavioral problems. Therefore, the assessment of in utero exposure to abused substance is extremely necessary and is relevant for the care of the mother and the offspring. In this sense, several approaches are possible; however, recently the evaluation of in utero exposure to abused drugs has been achieved by testing biological specimens coming from fetus or newborn, pregnant or nursing mother, or from both the fetus and the mother. Maternal and neonatal biological materials reflect exposure in a specific time period and each of them has different advantages and disadvantages in terms of accuracy, time window of exposure and cost/benefit ratio. The methodology for identification and determination of abused substances in biological materials are of great importance. Consequently, sensitive and specific bioanalytical methods are necessary to accurately measure biomarkers. Different immunoassays methods are used as screening methods for drug testing in the above reported specimens, however, the results from immunoassays should be carefully interpreted and confirmed by a more specific and sensitive chromatographic methods such as GC-MS or LC-MS. The interest in the development and optimization of analytical techniques to detect abused substances in different specimens is explained by the several possibilities and information that they can provide.


Suppression of c-myc and c-myb expression in myeloid cell lines treated with recombinant tumor necrosis factor-alpha.
Proto-oncogenes are thought to be involved in cellular differentiation and proliferation. Tumor necrosis factors (TNFs) are specific cytokines that have cytostatic and cytotoxic effects in vitro against a wide range of human tumor cells. We have previously demonstrated that recombinant TNFs (rTNFs) have an antiproliferative effect on certain human leukemic cell lines (HL-60, KBM3, KBM5) and no effect on others (K562). To study the possible role of the c-myc and c-myb oncogenes in this antiproliferative effect of TNF, we examined their expression in cell lines HL-60, KBM3, KBM5, and K562 before and after incubation with rTNF-alpha. Expression of c-myc and c-myb was elevated in all cell lines prior to incubation with rTNF-alpha. In the sensitive cell lines HL-60, KBM5, and KBM3 expression of c-myc and c-myb decreased rapidly 8-, 16-, and 4-fold, respectively, by 24 hr. K562 cells, insensitive to rTNF-alpha, exhibited no change in c-myc or c-myb expression over 24 hr. These studies demonstrated that down-regulation of c-myc and c-myb expression were associated with antiproliferative effects of rTNF-alpha on these cell lines.


Structural and mutational analysis of the ribosome-arresting human XBP1u.
XBP1u, a central component of the unfolded protein response (UPR), is a mammalian protein containing a functionally critical translational arrest peptide (AP). Here, we present a 3 Å cryo-EM structure of the stalled human XBP1u AP. It forms a unique turn in the ribosomal exit tunnel proximal to the peptidyl transferase center where it causes a subtle distortion, thereby explaining the temporary translational arrest induced by XBP1u. During ribosomal pausing the hydrophobic region 2 (HR2) of XBP1u is recognized by SRP, but fails to efficiently gate the Sec61 translocon. An exhaustive mutagenesis scan of the XBP1u AP revealed that only 8 out of 20 mutagenized positions are optimal; in the remaining 12 positions, we identify 55 different mutations increase the level of translational arrest. Thus, the wildtype XBP1u AP induces only an intermediate level of translational arrest, allowing efficient targeting by SRP without activating the Sec61 channel.


F-Met-Leu-Phe and echo 9 virus interaction with human granulocytes. Changes of cell membrane structure.
Biophysical and biochemical methods were applied for investigation of cell membrane properties of human polymorphonuclear leukocytes (PMNs) exposed to the chemotactic peptide N-formylmethionyl-leucylphenylalanine (f-Met-Leu-Phe) and echovirus type 9, strain A, Barty. Steady-state fluorescence depolarization with diphenylhexatriene demonstrated no gross changes of the total membrane fluidity under the different experimental conditions. However, by means of the monomer-excimer technique with pyrenedecanoic acid (PDA), significant changes of the local membrane structure were detected for both agents. As demonstrated by a higher excimer ratio, the membrane area available for the PDA molecules was restricted by f-Met-Leu-Phe. This effect was dependent on the dose and on the time of interaction of the chemotactic peptide. These experimental findings were explained by the formation of functional receptor units ("activated membrane"). Echo 9 virus exhibited the opposite effect, characterized by a higher ratio of monomers, which also depended on the viral dose and the time of virus-PMN interaction. These virus-induced findings were explained by the dissolution of functional receptor units. Consecutive exposure of the PMNs to f-Met-Leu-Phe and echovirus, or vice versa, demonstrated a virus-predominant effect on the membrane structures.


Vascular phases in imaging and their role in focal liver lesions assessment.
The incidental finding of a liver lesion with basic ultrasound is one of the most common clinical issues. Some of the liver lesions which present typical morphological B-mode features (e.g. cysts, typically localized focal fatty sparing/accumulations, hyperechoic hemangiomas) can be easily diagnosed by conventional ultrasound without the need of further diagnostic procedures. Others frequently necessitate further investigation with contrast-enhanced imaging techniques or biopsy in order to differentiate benign from malignant lesions and obtain a final diagnosis. This paper will discuss differences between vascular phases of different cross-sectional contrast-enhanced methods, as well as their subsequent benefits for focal liver lesions (FLLs) assessment, adding also a particular emphasis on small FLLs detection and characterization.


Driving factors of energy related CO2 emissions at a regional level in the residential sector of Iran.
Iran has increased its CO2 emissions significantly during the past few decades. The household sector in Iran contributes one of the largest sectors of CO2 emissions. Despite this significant contribution, the existing policies have predominantly concentrated on large-scale initiatives while overlooking the regional role in shaping and implementing these plans. Therefore, this study investigates the relationship between CO2 emissions and the efficient factors in three major groups including energy, climate, and household socio-economic factors. This study aims to address regional carbon emissions and develop CO2 reduction policies tailored to each region's specific circumstances. It focuses on planning strategies at the regional level to effectively tackle CO2 emissions. Household panel data of 28 provinces of Iran are employed by using both static and dynamic panel models for the years 2001 to 2019. Static estimation includes Fixed Effect (FE), Random Effect (RE) and pooled Partial least squares (PLS), Dynamic estimation includes difference Generalized Method of Moments (GMM) and system Generalized Method of Moments (GMM). The empirical result of the static method showed positive dependence of household CO2 emissions on Heating Degree Days (HDD), Cooling Degree Days (CDD), precipitation level, oil consumption, gas consumption, household income, size of household, and also building stocks. In more detail, educational rate, dummy variable (removal of energy subsidy), and oil price reveal the greatest negative impact on the emissions with elasticities of - 0.428, - 0.31, and - 0.15; It represents 1% increase causes - 0.428, - 0.31, - 0.15, decrease CO2 emissions, respectively. however, household size, gas consumption, and oil consumption show the most significant positive effects on CO2 emissions with 1 percent increase causes CO2 emissions increases by 0.1, 0.044, and 0.026, respectively. Regarding the impact of climate factors, a 1% increase in Heating Degree Days, Cooling Degree Days, and precipitation level causes CO2 emissions increase by 0.024%, 0.004%, and 0.011% respectively, due to an increase in fossil energy demand. Results of the dynamic method of the system Generalized Method of Moments are similar to the static estimation results, except for that household size and urbanization are not significant. Also, removing the energy subsidy for fossil fuels due to substantial subsidy in fossil fuels in Iran or implementing a re-pricing energy policy can be a beneficial way to control carbon emissions from households within the provinces of the country. However, it is important to consider that this shift could potentially transfer subsidies to investments in the private sector for renewable energies.


A bioluminescent Pseudomonas aeruginosa wound model reveals increased mortality of type 1 diabetic mice to biofilm infection.
To examine how bacterial biofilms, as contributing factors in the delayed closure of chronic wounds in patients with diabetes, affect the healing process. We used daily microscopic imaging and the IVIS Spectrum in vivo imaging system to monitor biofilm infections of bioluminescent Pseudomonas aeruginosa and evaluate healing in non-diabetic and streptozotocin-induced diabetic mice. Our studies determined that diabetes alone did not affect the rate of healing of full-depth murine back wounds compared with non-diabetic mice. The application of mature biofilms to the wounds significantly decreased the rate of healing compared with non-infected wounds for both non-diabetic as well as diabetic mice. Diabetic mice were also more severely affected by biofilms displaying elevated pus production, higher mortality rates and statistically significant increase in wound depth, granulation/fibrosis and biofilm presence. Introduction of a mutant Pseudomonas aeruginosa capable of producing high concentrations of cyclic di-GMP did not result in increased persistence in either diabetic or non-diabetic animals compared with the wild type strain. Understanding the interplay between diabetes and biofilms may lead to novel treatments and better clinical management of chronic wounds.


Transverse-plane mechanics at the knee and tibia in runners with and without a history of achilles tendonopathy.
Retrospective cohort study. To determine if runners with a history of Achilles tendonopathy (AT) demonstrate a difference in transverse-plane motion and moments at the distal tibia and knee compared to runners without a history of AT. One of the more prevalent overuse running injuries is AT. It has been hypothesized that prolonged pronation causes contradictory rotational forces acting on the AT. Conflicting joint motions may also accentuate the rotation of the tendon. This may further affect the opposing transverse-plane moments at the knee and distal tibia. Eight runners with a history of AT and a noninjured control group consisting of 8 runners ran along a 20-m runway at a fixed speed. Biomechanical measurements were taken using a 6-camera motion analysis system and a force plate. Student t tests were employed to determine statistically significant differences (P </= .05) in transverse-plane motion and moment variables at the distal tibia and knee between groups. The AT group showed less tibial external rotation moment (P = .01) and peak knee internal rotation (P = .05) compared to the control group. There was no difference in external rotation moment at the knee (P = .34) or peak tibial internal rotation (P = .44). Runners with a previous history of AT exhibited less tibial external rotation moments during running. The lack of control in the transverse-plane at the distal tibia may be due to decreased function of the muscles primarily responsible for transverse-plane motion, resulting in greater strain on the AT in the transverse-plane. Designing an exercise program to strengthen lower leg muscles and improve distal tibial control in the transverse-plane may reduce the risk for developing AT or augment the rehabilitation of AT.


Effect of acyclovir and interferon on human hematopoietic progenitor cells.
Continuous in vitro exposure of human bone marrow cells to acyclovir (approximately 200 microM) or human leukocyte interferon (approximately 250 U/ml) caused 50% inhibition of granulocyte colony-forming cell differentiation. Colonies expressed in the presence of either agent were reduced both in size and number. Erythroid progenitors were more resistant than granulocyte progenitors to the antiproliferative effects of acyclovir. Progenitor cells of patients recovering from cytotoxic chemotherapy were no more sensitive to the effects of acyclovir or interferon than were cells obtained from patients before chemotherapy.


[Effect of rosiglitazone on level of serum TNF-alpha and its relation with insulin resistance in type 2 diabetes mellitus].
To investigate the change of serum TNF-alpha level in type 2 diabetes mellitus after treating with rosiglitazone, and the relationship between TNF-alpha and insulin resistance. The level of serum TNF-alpha in 27 patients was tested by radioimmunoassay before and after treating with rosiglitazone. Free insulin, free blood sugar, TG, HDL-C and weight were measured, and HOMA-IR was calculated. The level of serum TNF-alpha in the patients was higher than that in the controls (P < 0.005) and rosiglitazone could decrease the level of serum TNF-alpha (P < 0.001). The level of serum TNF-alpha was positively related to serum TNF-alpha, TG, FBS and ln (HOMA-IR), and it was negatively related to HDL-C (r = -0.508, P < 0.05). Rosiglitazone can decrease the level of serum TNF-alpha significantly and improve insulin resistance.


Bitter sweeteners: tetrazole derivatives of arylsulfonylalcanoids--synthesis, structure and comparative study.
Within a research project aimed at the design of new sweeteners, the tetrazole moiety was introduced to arylsulfonylalkanoic acids (ASA) as a bioisostere of the carboxyl group. The crystal structures of four newly synthesized tetrazole derivatives and one intermediate product of the reaction were determined in order to explain the bitter taste of these compounds. Three chiral compounds crystallize as racemic mixtures in centrosymmetric space groups of the monoclinic system, whereas the non-chiral compound, with a higher dipole moment, crystallizes in the polar space group Cc. Intermolecular N-H...N hydrogen bonds between tetrazole moieties were observed in all four structures and are compared with the analogous interactions observed in tetrazole derivatives deposited in the Cambridge Structural Database (CSD). Specifically, the typical N1-H...N4 as well as N1-H...N3 interactions, which are less abundant in the CSD, are described. The formation of the latter interaction type can be hypothetically explained by an asymmetry of pi-electron distribution in the tetrazole rings caused by the crystalline environment. Important features of the crystal architecture are the chains of molecules linked by N-H...N bonds. A possible reason for the lack of a sweet taste of the tetrazoles investigated may be the improper position of the tetrazole H atom, and the mutual orientation of the proton donor and acceptor in their molecules. This orientation does not allow the tetrazoles to interact with the sweet-taste receptor in a way similar to that of ASA. The bitter taste of the investigated compounds needs further study.


[Heart failure patient management: evolution, organization, application on a local scale].
Heart failure is a severe disease which prevalence and incidence are steadily growing, with high morbidity and mortality rates, and responsible for significant financial cost. This evolution requires our therapeutic approach to be modified, by favouring multidisciplinary management and including patient education. The proof of the effectiveness of these measures is brought by many studies, and therapeutic education is now recommended by the European Society of Cardiology. This new approach of management is set out according to various procedures. In Le Creusot, the therapeutic unit of heart failure management was created in September 2004 and operate on a day hospital pattern. The results are satisfactory knowing that the average duration of spells fell by 33% after one year. Further local initiatives are spreading and need shall arise to pooling equipment and personnel as well as reinforcing collaboration between public and private professionals in order to improve heart failure patients' management. In the meantime a more sustained involvement of the authorities is to be hoped.


Accommodation and T-independent B cell tolerance in rats with long term surviving hamster heart xenografts.
It was previously reported that treatment with leflunomide (LF; 10 mg/kg/day) together with cyclosporine (CsA; 10 mg/kg/day) resulted in long term survival of hamster heart xenografts (Xg) in rats and that LF could be withdrawn 2 to 4 wk after transplantation. To study the mechanisms allowing withdrawal of LF, second hamster heart Xgs were transplanted 6 wk after the first xenograft. Only the rats that received LF for 4 wk accepted second Xgs (>30 days; n = 5). Hence, after 4 wk of LF, the rats developed partial B cell tolerance, as they were unable to produce T-independent (CsA-resistant) XAbs. Rejection of second Xgs (2-4 days; n = 5) in the 2-wk LF group resulted in the formation of IgM xenoantibodies (XAbs) localizing together with complement within rejected grafts. However, these XAbs did not affect first Xgs, suggesting that the latter Xgs became resistant to this IgM XAb-mediated rejection, a phenomenon referred to as accommodation. Accommodation was further confirmed as adoptive transfer of IgM XAbs, which resulted in hyperacute Xg rejection in naive rats (<1 h; n = 5), did not cause rejection in long term survivors (>30 days; n = 4). This was associated with a down-regulation of the expression on the graft endothelial cells of adhesion molecules (believed to be important expressers of xenogeneic epitopes), such as P- and E-selectins. Interestingly, these adhesion molecules reappeared after retransplanting the accommodated Xgs to naive recipients. In conclusion, depending on the duration of the LF treatment, long term survival of hamster hearts in CsA-treated rats is based in part on accommodation and in part on T-independent B cell tolerance.


Evaluating Sustainable Development Pathways for Protein- and Peptide-Based Bioadsorbents for Phosphorus Recovery from Wastewater.
Recovering phosphate (P) from point sources such as wastewater effluent is a priority in order to alleviate the impacts of eutrophication and implement a circular economy for an increasingly limited resource. Bioadsorbents featuring P-binding proteins and peptides offer exquisite P specificity and sensitivity for achieving ultralow P concentrations, i.e., <100 μg P L-1, a discharge limit that has been implemented in at least one treatment facility in nine U.S. states. To prioritize research objectives for P recovery in wastewater treatment, we compared the financial and environmental sustainability of protein/peptide bioadsorbents to those of LayneRT anion exchange resin. The baseline scenario (reflecting lab-demonstrated performance at a full-scale implementation) had costs that were 3 orders of magnitude higher than those for typical wastewater treatment. However, scenarios exploring bioadsorbent improvements, including increasing the P-binding capacity per unit volume by using smaller P-selective peptides and nanoparticle base materials and implementing reuse, dramatically decreased median impacts to $1.06 m-3 and 0.001 kg CO2 equiv m-3; these values are in line with current wastewater treatment impacts and lower than the median LayneRT impacts of $4.04 m-3 and 0.19 kg CO2 equiv m-3. While the financial viability of capturing low P concentrations is a challenge, incorporating the externalities of environmental impacts may provide a feasible path forward to motivate ultralow P capture.


Glucose Deprivation Enhances the Antiproliferative Effects of Oral Hypoglycemic Biguanides in Different Molecular Subtypes of Breast Cancer: an in Vitro Study.
Extensive in vitro studies have been conducted to evaluate the anticancer activity of oral hypoglycemic agents. Many of these studies experienced detrimental limitations, since they were conducted on cancer cells commonly grown in culture media consisting of extremely high concentrations of growth factors and glucose. The present study was aimed at exploring the antiproliferative effects of the commonly studied metformin and the less frequently reported phenformin oral hypoglycemic agents on different molecular subtypes of breast cancer under rich glucose and glucose deprived conditions. Our results indicate that under glucose deprived conditions, which better reflect the factual glucose-starved solid tumors in vivo, biguanides exert more antiproliferative activities against the three molecular subtypes of breast cancer cell lines examined in this study. In addition, the observed antiproliferative activities of biguanides appear to be mediated by apoptosis induction in breast cancer cells. This induction is significantly augmented under glucose deprived conditions.


A study of the physical and chemical properties of four resin composite luting cements.
This study evaluated the surface microhardness and flexural and compressive strengths of five luting cements and compared the degree of conversion of dual and autopolymerized forms of four resin-based luting cements. Four resin composite luting cements-Panavia F, Variolink 2, RelyX Unicem Applicap, and RelyX ARC-and a polycarboxylate cement (Durelon, control group) were used in three-point bending, compression, and Vickers hardness tests following water storage for 1 week. Resin composite cements were additionally investigated with both dual and autopolymerization techniques under Fourier transformed infrared spectroscopy. Differences were analyzed using one-way ANOVA. The highest flexural strengths were obtained with Variolink 2 (90 MPa, SD 22), whereas the lowest were observed with Durelon (28 MPa, SD 4). RelyX Unicem showed the highest hardness values (44 HV, SD 5), whereas Variolink 2 gave the lowest (32 HV, SD 6). The highest compressive strengths were obtained with RelyX Unicem (145 MPa, SD 32), whereas the lowest were observed with Durelon (41 MPa, SD 17). For both dual and autopolymerized groups, RelyX ARC showed the highest degrees of conversion (81% and 61%, respectively) and RelyX Unicem had the lowest (56% and 26%, respectively). Resin composite luting cements of similar chemical characterizations differed in their physical properties, and polymerization method influenced their degree of conversion.


Downregulation of lncRNA FOXD2-AS1 Confers Radiosensitivity to Gastric Cancer Cells via miR-1913/SETD1A Axis.
Long noncoding RNA FOXD2 adjacent opposite strand RNA1 (FOXD2-AS1) plays an oncogenic role in various cancers, including gastric cancer (GC). However, the function of FOXD2-AS1 in regulating radiosensitivity of GC cells and its underlying molecular mechanisms have not been elucidated. This study aimed to figure out the potential mechanisms of FOXD2-AS1 in regulating GC cell radiosensitivity. RT-qPCR revealed upregulation of FOXD2-AS1 in GC cells exposed to radiation. Subcellular fractionation assay was used to localize FOXD2-AS1 in GC cells. Colony formation, MTT, EdU, and flow cytometry assays were performed to investigate the role of FOXD2-AS1 in regulating cell proliferation, cell cycle progression, and cell apoptosis. Western blotting was used to assess protein levels of apoptosis-associated markers and SET domain containing 1A (SETD1A). Homologous recombination reporter assay was conducted to explore the effect of FOXD2-AS1 on DNA damage repair. The downstream molecules of FOXD2-AS1 were identified with RNA pulldown, luciferase reporter, and RNA immunoprecipitation assays. The results showed that FOXD2-AS1 knockdown suppressed cell proliferation and cell cycle progression and promoted cell apoptosis and radiosensitivity of GC. FOXD2-AS1 could bind with miR-1913 in GC cells. In addition, miR-1913 targeted SETD1A, which was highly expressed in GC cells. Overexpression of SETD1A reversed FOXD2-AS1 silencing-induced effects on proliferation, apoptosis, and radiosensitivity of GC cells. In conclusion, knocking down FOXD2-AS1 enhances the radiosensitivity of GC cells by sponging miR-1913 to upregulate SETD1A expression.


Regulation of autoimmune arthritis by the SHP-1 tyrosine phosphatase.
The Src homology region 2 domain-containing phosphatase-1 (SHP-1) is known to exert negative regulatory effects on immune cell signaling. Mice with mutations in the Shp1 gene develop inflammatory skin disease and autoimmunity, but no arthritis. We sought to explore the role of SHP-1 in arthritis using an autoimmune mouse model of rheumatoid arthritis. We generated Shp1 transgenic (Shp1-Tg) mice to study the impact of SHP-1 overexpression on arthritis susceptibility and adaptive immune responses. SHP-1 gene and protein expression as well as tyrosine phosphatase activity were evaluated in spleen cells of transgenic and wild type (WT) mice. WT and Shp1-Tg (homozygous or heterozygous for the transgene) mice were immunized with human cartilage proteoglycan (PG) in adjuvant, and arthritis symptoms were monitored. Protein tyrosine phosphorylation level, net cytokine secretion, and serum anti-human PG antibody titers were measured in immune cells from WT and Shp1-Tg mice. WT mice were treated with regorafenib orally to activate SHP-1 either before PG-induced arthritis (PGIA) symptoms developed (preventive treatment) or starting at an early stage of disease (therapeutic treatment). Data were statistically analyzed and graphs created using GraphPad Prism 8.0.2 software. SHP-1 expression and tyrosine phosphatase activity were elevated in both transgenic lines compared to WT mice. While all WT mice developed arthritis after immunization, none of the homozygous Shp1-Tg mice developed the disease. Heterozygous transgenic mice, which showed intermediate PGIA incidence, were selected for further investigation. We observed differences in interleukin-4 and interleukin-10 production in vitro, but serum anti-PG antibody levels were not different between the genotypes. We also found decreased tyrosine phosphorylation of several proteins of the JAK/STAT pathway in T cells from PG-immunized Shp1-Tg mice. Regorafenib administration to WT mice prevented the development of severe PGIA or reduced disease severity when started after disease onset. Resistance to arthritis in the presence of SHP-1 overexpression likely results from the impairment of tyrosine phosphorylation (deactivation) of key immune cell signaling proteins in the JAK/STAT pathway, due to the overwhelming tyrosine phosphatase activity of the enzyme in Shp1-Tg mice. Our study is the first to investigate the role of SHP-1 in autoimmune arthritis using animals overexpressing this phosphatase. Pharmacological activation of SHP-1 might be considered as a new approach to the treatment of autoimmune arthritis.


Initial experience with a multiple parallel guidewire support system for complex tortuous aortic arch navigation and great vessel catheterization: technical note.
The ability to traverse an anatomically challenging and complex arch is paramount to the success of any neuroendovascular procedure. With age, the aortic arch becomes elongated, calcified, and less compliant. The authors present the initial experience with a multiple parallel guidewire system (ZigiWire Mode 3) for catheterization through a complex tortuous aortic arch to access extracranial vessels. The ZigiWire is an organized guidewire system that uses consecutive delivery of 3 small-diameter (0.014-inch) guidewires that are progressively advanced in parallel to secure support-wire access. The authors have found it useful in situations in which traditional methods for great-vessel access have failed. Moreover, the progressive construction of a large wire from smaller wires prevents "kickback" force from a single larger guidewire, allowing stable distal access. The authors have been able to advance different diagnostic and guide catheters over the ZigiWire. This guidewire has allowed them to successfully complete neuroendovascular procedures in patients who were previously considered unsuitable for the procedure because of tortuous vascular access.


Rare scleroderma autoantibodies to the U11 small nuclear ribonucleoprotein and to the trimethylguanosine cap of U small nuclear RNAs.
We have identified a scleroderma serum (Ru) with a previously undescribed specificity to protein components of the U11 small nuclear ribonucleoprotein particle (snRNP), a low-abundance member of the Sm class of U RNPs. The U11 RNP can be specifically immunoprecipitated from sonicated HeLa cells with Ru serum. In nuclear extracts, a fraction of the U11 particle is found complexed to the U12 RNP, an even lower abundance Sm snRNP. In glycerol gradient fractions, Ru serum identifies a 65-kDa protein that cosediments with the U11-U12 complex and is shifted upon targeted degradation of the U12 RNA. The 65-kDa protein therefore appears to be a component of the U11-U12 snRNP complex, whereas another Ru-reactive (140 kDa) protein may be associated with the free U11 RNP. The Ru serum also contains autoantibodies directed against the trimethylguanosine cap of U RNAs. This rare specificity has been described previously in only three other scleroderma patients.


Professional, practice and political issues in the history of New Zealand's remote rural 'backblocks' nursing: the case of Mokau, 1910-1940.
The new role of 'backblocks' nursing, established in 1909 to provide a nursing, midwifery, emergency and public health service to New Zealand's remote rural regions, created opportunities and challenges for the profession. For three decades, the novel nature of the role also provided numerous stakeholders with the opportunity to contest their authority and influence. This article explores the professional, practice and political issues of backblocks nursing through a case study of Mokau, a remote rural community in the North Island of New Zealand, 1910-1940. In particular, it considers professional issues of recruitment and retention, practice issues in delivering the new service in a challenging environment with few resources, and political issues in defining the scope of nurses' practice and dealing with competing stakeholders keen to determine its potential and limits. These issues were exacerbated by the location of the Mokau district in two administrative health regions.


Quantitating the Transfer of the HTLV-1 p8 Protein Between T-Cells by Flow Cytometry.
The Human T-cell leukemia virus type 1 (HTLV-1)-encoded accessory protein p8 is cleaved from the precursor protein p12 encoded by the HTLV-1 open reading frame I. Both p12 and p8 are thought to contribute to efficient viral persistence. Mechanistically, p8 induces T-cell conjugates and cellular conduits. The latter are considered to facilitate transfer of p8 to target cells and virus transmission. Transfer of p8 between p8-expressing T-cells and recipient cells has been analyzed by immunofluorescence and live imaging. However, automatic quantitation of p8-transfer between cells has not been studied yet. Here we developed a novel method allowing time saving quantitation of p8 transfer between cells by flow cytometry. After establishing a protocol for the detection of intracellular p8 by flow cytometry and validation of p8 protein expression by western blot and immunofluorescence, we set up a co-culture assay between p8-expressing donor Jurkat T-cells and recipient Jurkat T-cells that had been prestained with a well-retained live cell dye. Upon quantitating the amount of p8 positive recipient cells with regard to the percentage of p8 expressing donor cells, time course experiments confirmed that p8 is rapidly transferred between Jurkat T-cells. We found that p8 enters approximately 5% of recipient T-cells immediately upon co-culture for 5 min. Prolonged co-culture for up to 24 h revealed an increase of relative p8 transfer to approximately 23% of the recipient cells. Immunofluorescence analysis of co-culture experiments and manual quantitation of p8 expression in fluorescence images confirmed the validity of the flow cytometry based assay. Application of the new assay revealed that manipulation of actin polymerization significantly decreased p8 transfer between Jurkat T-cells suggesting an important role of actin dynamics contributing to p8 transfer. Further, transfer of p8 to co-cultured T-cells varies between different donor cell types since p8 transfer could hardly been detected in co-cultures of 293T donor cells with Jurkat acceptor cells. In summary, our novel assay allows automatic and rapid quantitation of p8 transfer to target cells and might thus contribute to a better understanding of cellular processes and dynamics regulating p8 transfer and HTLV-1 transmission.


Redox Control of Aphid Resistance through Altered Cell Wall Composition and Nutritional Quality.
The mechanisms underpinning plant perception of phloem-feeding insects, particularly aphids, remain poorly characterized. Therefore, the role of apoplastic redox state in controlling aphid infestation was explored using transgenic tobacco (Nicotiana tabacum) plants that have either high (PAO) or low (TAO) ascorbate oxidase (AO) activities relative to the wild type. Only a small number of leaf transcripts and metabolites were changed in response to genotype, and cell wall composition was largely unaffected. Aphid fecundity was decreased significantly in TAO plants compared with other lines. Leaf sugar levels were increased and maximum extractable AO activities were decreased in response to aphids in all genotypes. Transcripts encoding the Respiratory Burst Oxidase Homolog F, signaling components involved in ethylene and other hormone-mediated pathways, photosynthetic electron transport components, sugar, amino acid, and cell wall metabolism, were increased significantly in the TAO plants in response to aphid perception relative to other lines. The levels of galactosylated xyloglucan were decreased significantly in response to aphid feeding in all the lines, the effect being the least in the TAO plants. Similarly, all lines exhibited increases in tightly bound (1→4)-β-galactan. Taken together, these findings identify AO-dependent mechanisms that limit aphid infestation.


Implicit learning of color-word associations using a Stroop paradigm.
Our experiments asked whether implicit learning occurs for novel nonverbal associations. We presented subjects with color names printed in incongruent colors; subjects were asked to name the color in which the word was printed. In Experiment 1, each of 7 color words were associated with the same incongruent color across 6 blocks of trials, and then the color-word associations were abruptly changed. Both control subjects and patients with amnesia reduced their color-naming times across the first 6 trial blocks, and naming times increased when the color-word associations were changed. In Experiment 2, similar results were obtained when neutral words were associated with colors. In Experiment 3, we found that naming times were not disrupted when an irrelevant dimension (typecase) was changed. Finally, in Experiment 4, we found that the effect persisted across a 5-min delay. These studies provide evidence that implicit learning occurs for nonverbal associations and is independent of the brain structures damaged in amnesia.


[Pathogenesis and drug therapy of acute laminitis in horses: a literature review].
Impairment of the microcirculation in the distal part of the foot is an important factor in the aetiology of equine laminitis. To prevent the development of chronic laminitis and the resulting complications, two new therapeutical principles were tested in the treatment of acute laminitis. Thrombosis which frequently occurs in the corium of the foot, could be controlled by administration of anticoagulants such as heparin. Alpha-receptor blocking agents such as phenoxybenzamine were used, to improve the peripheral blood flow. The use of these drugs and that of anti-inflammatory agents during the onset of acute laminitis is discussed in this review paper.


The performance of aminoalkyl/fluorocarbon/hydrocarbon-modified xerogel coatings against the marine alga Ectocarpus crouaniorum: relative roles of surface energy and charge.
The effect of a series of xerogel coatings modified with aminoalkyl/fluorocarbon/hydrocarbon groups on the adhesion of a new test species, the filamentous brown alga Ectocarpus crouaniorum, has been explored, and compared with the green alga Ulva linza. The results showed that E. crouaniorum adhered weakly to the less polar, low wettability coatings in the series, but stronger adhesion was shown on polar, higher surface energy coatings containing aminoalkyl groups. The results from a separate series of coatings tuned to have similar surface energies and polarities after immersion in artificial seawater (ASW), but widely different surface charges, demonstrated that surface charge was more important than surface energy and polarity in determining the adhesion strength of both E. crouaniorum and U. linza on xerogel coatings. No correlation was found between adhesion and contact angle hysteresis. X-ray photoelectron spectroscopy analysis of samples after immersion in ASW confirmed the presence of charged ammonium groups on the surface of the aminoalkylated coatings.


TMEM106B Puncta Is Increased in Multiple Sclerosis Plaques, and Reduced Protein in Mice Results in Delayed Lipid Clearance Following CNS Injury.
During inflammatory, demyelinating diseases such as multiple sclerosis (MS), inflammation and axonal damage are prevalent early in the course. Axonal damage includes swelling, defects in transport, and failure to clear damaged intracellular proteins, all of which affect recovery and compromise neuronal integrity. The clearance of damaged cell components is important to maintain normal turnover and restore homeostasis. In this study, we used mass spectrometry to identify insoluble proteins within high-speed/mercaptoethanol/sarcosyl-insoluble pellets from purified white matter plaques isolated from the brains of individuals with relapsing-remitting MS (RRMS). We determined that the transmembrane protein 106B (TMEM106B), normally lysosome-associated, is insoluble in RRMS plaques relative to normal-appearing white matter from individuals with Alzheimer's disease and non-neurologic controls. Relative to wild-type mice, hypomorphic mice with a reduction in TMEM106B have increased axonal damage and lipid droplet accumulation in the spinal cord following myelin-oligodendrocyte-glycoprotein-induced experimental autoimmune encephalomyelitis. Additionally, the corpora callosa from cuprizone-challenged hypomorphic mice fail to clear lipid droplets efficiently during remyelination, suggesting that when TMEM106B is compromised, protein and lipid clearance by the lysosome is delayed. As TMEM106B contains putative lipid- and LC3-binding sites, further exploration of these sites is warranted.


Cellular immune defects to Epstein-Barr virus-determined antigens in young males.
Three males with the X-linked lymphoproliferative syndrome (XLP) with hypo- or agammaglobulinemia following Epstein-Barr virus (EBV) infection and two males with the chronic mononucleosis syndrome were investigated for immune responses to EBV-determined antigens. Males with XLP showed profound cellular immune defects. Markedly diminished responses of natural killer cell and interferon-activated killer cell activities and impaired leukocyte migration inhibition responses to phytohemagglutinin were determined in patients with XLP. The two patients with chronic mononucleosis showed less severe defects. All patients showed partial or complete impairment of their EBV-specific immune responses as measured by leukocyte migration inhibition. EBV-specific antibodies were markedly diminished against EBV-associated nuclear antigen, early antigen, and viral capsid antigen in males with XLP. In contrast, patients with chronic mononucleosis had elevated antibodies to most EBV-specific antigens. Individuals with life-threatening EBV-induced lymphoproliferative disorders may exhibit multiple defective immune mechanisms against the virus.


Structural and physiological analyses in Salsoleae (Chenopodiaceae) indicate multiple transitions among C3, intermediate, and C4 photosynthesis.
In subfamily Salsoloideae (family Chenopodiaceae) most species are C4 plants having terete leaves with Salsoloid Kranz anatomy characterized by a continuous dual chlorenchyma layer of Kranz cells (KCs) and mesophyll (M) cells, surrounding water storage and vascular tissue. From section Coccosalsola sensu Botschantzev, leaf structural and photosynthetic features were analysed on selected species of Salsola which are not performing C4 based on leaf carbon isotope composition. The results infer the following progression in distinct functional and structural forms from C3 to intermediate to C4 photosynthesis with increased leaf succulence without changes in vein density: From species performing C3 photosynthesis with Sympegmoid anatomy with two equivalent layers of elongated M cells, with few organelles in a discontinuous layer of bundle sheath (BS) cells (S. genistoides, S. masenderanica, S. webbii) > development of proto-Kranz BS cells having mitochondria in a centripetal position and increased chloroplast number (S. montana) > functional C3-C4 intermediates having intermediate CO2 compensation points with refixation of photorespired CO2, development of Kranz-like anatomy with reduction in the outer M cell layer to hypodermal-like cells, and increased specialization (but not size) of a Kranz-like inner layer of cells with increased cell wall thickness, organelle number, and selective expression of mitochondrial glycine decarboxylase (Kranz-like Sympegmoid, S. arbusculiformis; and Kranz-like Salsoloid, S. divaricata) > selective expression of enzymes between the two cell types for performing C4 with Salsoloid-type anatomy. Phylogenetic analysis of tribe Salsoleae shows the occurrence of C3 and intermediates in several clades, and lineages of interest for studying different forms of anatomy.


Combination of the advantages of chromatographic methods based on active components for the quality evaluation of licorice.
A rapid, improved and comprehensive method including high-performance thin-layer chromatography, fingerprint technology and single standard to determine multiple components was developed and validated for the quality evaluation of licorice. In this study, a newly developed high-performance thin-layer chromatography method was first used for authentication of licorice, which achieved simultaneous identification of multiple bands including five bands for known bioactive components by comparing their retention factor values and colors with the standards. For fingerprint analysis, 8 of 16 common peaks were identified. Simultaneously, similarity analysis which showed very similar patterns and hierarchical clustering analysis were performed to discriminate and classify the 27 batches of samples. Additionally, the single standard to determine multiple components method was first successfully achieved to quantify the eight important active markers in licorice including liquiritin apioside, liquiritin, isoliquiritin apioside, isoliquritin, neoisoliquiritin, liquiritigenin, isoliquiritigenin and glycyrrhizic acid. The easily available glycyrrhizic acid was selected as the reference substance to calculate relative response factors. Compared with the normal external standard method, this alternative method can be used to determine the multiple indices effectively and accurately. The validation result showed that the developed method was specific, accurate, precise, robust and reliable for the overall quality assessment of licorice.


Developmental changes in muscarinic receptor-stimulated phosphoinositide metabolism in rat brain.
Muscarinic receptor-stimulated phosphoinositide hydrolysis was investigated in rat brain during ontogeny by measuring the accumulation of [3H]inositol phosphates ([3H]InsPs) in cerebral cortex slices at various ages. Experiments with carbachol and acetylcholine showed that [3H]InsPs accumulation was maximal in 7-day-old rats (1477 +/- 98% of basal) and lowest in adult (75 days) rats (428 +/- 24% of basal). No differences were found in the EC50 values for both cholinergic agonists. This effect appeared to be mediated by the M1-muscarinic receptor subtype as it was blocked by pirenzepine with Ki = 29.1 +/- 7.1 nM (adults) and 87.9 +/- 18.2 nM (7-day-old rats). Incorporation of [3H]inositol into phospholipid decreased from day 3 to adulthood; however, when data of [3H]InsPs release were corrected for the incorporation at a given age, the highest stimulation by cholinergic agonists was still observed in 7-day-old rats. Among the other neurotransmitters tested (norepinephrine, histamine and serotonin), all known to stimulate phosphoinositide metabolism, none had the same developmental profile of [3H]InsPs accumulation as cholinergic agonists. In contrast to carbachol- and acetylcholine-stimulated phosphoinositide hydrolysis, the density of muscarinic binding sites, measured by [3H]quinuclidinyl benzilate binding, increased from day 3 to day 75. Acetylcholinesterase activity also increased during development. The dissociation of receptor binding sites from receptor-stimulated phosphoinositide metabolism suggests the presence of a more effective receptor-effector coupling at specific times of neonatal development, particularly 1 week. Furthermore, the fact that maximal stimulation of phosphoinositide hydrolysis coincides with the period of brain growth spurt in the rats suggests that this system in the cerebral cortex might be involved in the processes of cell division and differentiation.


A New Perspective on the Risk of Hypercoagulopathy in Ovarian Hyperstimulation Syndrome Using Thromboelastography.
Thromboelastography (TEG) provides real-time assessment of hemostasis by measuring the viscoelastic properties, coagulation factor, and platelet activity in whole blood samples. In this prospective case-control study, we wanted to investigate whether blood clot formation assessment, using TEG, can identify a hypercoagulable state in women with severe ovarian hyperstimulation syndrome (OHSS). Thirty-six women who were hospitalized with severe OHSS were allocated to the OHSS group and 32 women undergoing controlled ovarian hyperstimulation but who did not develop OHSS were allocated to the control group. The TEG indices were compared between women with severe OHSS and controls. All the coagulation indices assessed by TEG were significantly different in the OHSS group compared to the controls, depicting a hypercoagulable state. Median coagulation index was 3.6 (interquartile range: 2.80-4.15) and 1.45 (interquartile range: 0.20-2.30) in study group and controls, respectively ( P < .001). Our results show that TEG can be used to depict a hypercoagulable state in women with severe OHSS.


Adjuvant chemotherapy in elderly women with breast cancer (AChEW): an observational study identifying MDT perceptions and barriers to decision making.
As few older women with breast cancer receive adjuvant chemotherapy, we examined the barriers and perceptions of 24 UK NHS multidisciplinary breast cancer teams to offering this treatment to women ≥70 years. Questionnaires regarding 803 patients with newly diagnosed breast cancer were completed by specialist teams following discussion or outpatient consultation. Of 803 patients, 116 (14%), all <85 years, were offered chemotherapy and 66 (8%) received it. Only 94 of 309 (30%) of women with high-risk disease were offered chemotherapy, and 53 (17%) received it. The most common reasons for not offering chemotherapy were 'other treatments more appropriate' (usually patients with ER-positive tumours) or 'benefits too small' (63% and 54% of patients, respectively). Co-morbidities and frailty were less common reasons but became more frequent with increasing age. Recommendations regarding chemotherapy were made in the absence of documented HER2 and performance status in 29% and 33%, respectively. Treatment offered varied considerably between cancer centres. National guidelines need development describing the minimally acceptable data for decision making, incorporating objective fitness measures and specific treatment recommendations. Such guidelines will require educational support for implementation but should standardise care and improve chemotherapy uptake in this increasing population of older patients.


A 13-Gene Metabolic Prognostic Signature Is Associated With Clinical and Immune Features in Stomach Adenocarcinoma.
Patients with advanced stomach adenocarcinoma (STAD) commonly show high mortality and poor prognosis. Increasing evidence has suggested that basic metabolic changes may promote the growth and aggressiveness of STAD; therefore, identification of metabolic prognostic signatures in STAD would be meaningful. An integrative analysis was performed with 407 samples from The Cancer Genome Atlas (TCGA) and 433 samples from Gene Expression Omnibus (GEO) to develop a metabolic prognostic signature associated with clinical and immune features in STAD using Cox regression analysis and least absolute shrinkage and selection operator (LASSO). The different proportions of immune cells and differentially expressed immune-related genes (DEIRGs) between high- and low-risk score groups based on the metabolic prognostic signature were evaluated to describe the association of cancer metabolism and immune response in STAD. A total of 883 metabolism-related genes in both TCGA and GEO databases were analyzed to obtain 184 differentially expressed metabolism-related genes (DEMRGs) between tumor and normal tissues. A 13-gene metabolic signature (GSTA2, POLD3, GLA, GGT5, DCK, CKMT2, ASAH1, OPLAH, ME1, ACYP1, NNMT, POLR1A, and RDH12) was constructed for prognostic prediction of STAD. Sixteen survival-related DEMRGs were significantly related to the overall survival of STAD and the immune landscape in the tumor microenvironment. Univariate and multiple Cox regression analyses and the nomogram proved that a metabolism-based prognostic risk score (MPRS) could be an independent risk factor. More importantly, the results were mutually verified using TCGA and GEO data. This study provided a metabolism-related gene signature for prognostic prediction of STAD and explored the association between metabolism and the immune microenvironment for future research, thereby furthering the understanding of the crosstalk between different molecular mechanisms in human STAD. Some prognosis-related metabolic pathways have been revealed, and the survival of STAD patients could be predicted by a risk model based on these pathways, which could serve as prognostic markers in clinical practice.


Developments in Glycopeptide Antibiotics.
Glycopeptide antibiotics (GPAs) are a key weapon in the fight against drug resistant bacteria, with vancomycin still a mainstream therapy against serious Gram-positive infections more than 50 years after it was first introduced. New, more potent semisynthetic derivatives that have entered the clinic, such as dalbavancin and oritavancin, have superior pharmacokinetic and target engagement profiles that enable successful treatment of vancomycin-resistant infections. In the face of resistance development, with multidrug resistant (MDR) S. pneumoniae and methicillin-resistant Staphylococcus aureus (MRSA) together causing 20-fold more infections than all MDR Gram-negative infections combined, further improvements are desirable to ensure the Gram-positive armamentarium is adequately maintained for future generations. A range of modified glycopeptides has been generated in the past decade via total syntheses, semisynthetic modifications of natural products, or biological engineering. Several of these have undergone extensive characterization with demonstrated in vivo efficacy, good PK/PD profiles, and no reported preclinical toxicity; some may be suitable for formal preclinical development. The natural product monobactam, cephalosporin, and β-lactam antibiotics all spawned multiple generations of commercially and clinically successful semisynthetic derivatives. Similarly, next-generation glycopeptides are now technically well positioned to advance to the clinic, if sufficient funding and market support returns to antibiotic development.


δ15N Values in Crassostrea virginica Shells Provides Early Direct Evidence for Nitrogen Loading to Chesapeake Bay.
Crassostrea virginica is one of the most common estuarine bivalves in the United States' east coast and is frequently found in archaeological sites and sub-fossil deposits. Although there have been several sclerochronological studies on stable carbon and oxygen isotopes in the shells of this species, less is known about δ15N values within their shells, which could be a useful paleoenvironmental proxy to assess estuarine nitrogen dynamics. Modern C. virginica samples were collected in Chesapeake Bay for comparison with archaeological shells from nearby sites ranging in age from ~100 to 3,200 years old. Left valves were sampled by milling the hinge area and the resulting powder was analyzed for %N and δ15N values. Comparison of δ15N values between C. virginica shells shows relatively constant values from ~1250 BC to ~1800 AD. After ~1800 AD, there are rapid increases in 15N enrichment in the shells, which continue to increase in value up to the modern shell values. The increase in δ15N values is evidence of early anthropogenic impact in Chesapeake Bay. These results corroborate the observation that coastal nitrogen pollution occurred earlier than the 19th century and support the use of oyster shell δ15N values as a useful environmental proxy.


Roles of macrophage migration inhibitory factor in cartilage tissue engineering.
To obtain stable outcomes in regenerative medicine, understanding and controlling immunological responses in transplanted tissues are of great importance. In our previous study, auricular chondrocytes in tissue-engineered cartilage transplanted in mice were shown to express immunological factors, including macrophage migration inhibitory factor (MIF). Since MIF exerts pleiotropic functions, in this study, we examined the roles of MIF in cartilage regenerative medicine. We made tissue-engineered cartilage consisting of auricular chondrocytes of C57BL/6J mouse, atellocollagen gel and a PLLA scaffold, and transplanted the construct subcutaneously in a syngeneic manner. Localization of MIF was prominent in cartilage areas of tissue-engineered cartilage at 2 weeks after transplantation, though it became less apparent by 8 weeks. Co-culture with RAW264 significantly increased the expression of MIF in chondrocytes, suggesting that the transplanted chondrocytes in tissue-engineered cartilage could enhance the expression of MIF by stimulation of surrounding macrophages. When MIF was added in the culture of chondrocytes, the expression of type II collagen was increased, indicating that MIF could promote the maturation of chondrocytes. Meanwhile, toluidine blue staining of constructs containing wild type (Mif+/+) chondrocytes showed increased metachromasia compared to MIF-knockout (Mif-/-) constructs at 2 weeks. However, this tendency was reversed by 8 weeks, suggesting that the initial increase in cartilage maturation in Mif+/+ constructs deteriorated by 8 weeks. Since the Mif+/+ constructs included more iNOS-positive inflammatory macrophages at 2 weeks, MIF might induce an M1 macrophage-polarized environment, which may eventually worsen the maturation of tissue-engineered cartilage in the long term.


Predictors of Inconsistent Condom Use among Female Sex Workers: A Community-Based Study in a Red-Light Area of Kolkata, India.
Condom is an integral part of all interventions for female sex workers (FSWs). However, certain situations hinder them from practicing consistent condom use. This paper aims at identifying the situations that predict inconsistent condom use by FSWs of Sonagachi red-light area, Kolkata. The cross-sectional community-based observational study was conducted among 296 brothel-based FSWs of Sonagachi. Outcome variable, that is, inconsistent condom use was said to be present if any sex act with any type of partner was not protected by condom. Association with sociodemographic, occupational, and behavioral characteristics of FSWs was examined. Inconsistent condom use was present among 37.5% of the study participants, and this was predicted by the presence of a nonpaying partner (NPP) (adjusted odds ratio [AOR] [95% confidence interval (CI)]: 15.04 [7.52-30.08]), violence (AOR [95% CI]: 2.08 [1.07-4.03]) and sexual intercourse under the influence of alcohol (AOR [95% CI]: 1.86 [1.02-3.39]). The major cause behind nonuse of condom as reported by the participants was trust on partner. NPPs need to be given emphasis in program strategies for FSWs. The FSWs should constantly be motivated for safe sex emphasizing the fact that trust on partner may be detrimental for their health.


Converting a cysteine-rich natively noncatalytic protein to an artificial hydrogenase.
An artificial hydrogenase is constructed when the natively noncatalytic α-domain of the Cys-rich protein metallothionein (MT) is assembled with NiII. αMT binds four eq. of NiII in a non-cooperative manner where the addition of the 1st NiII eq. affords the most catalytically active species with little effect on photocatalytic H2 production during subsequent metal addition. The critical role of protonated Cys residue(s) in H-H bond formation is demonstrated.


Rare benign bone tumors and dystrophy in the hand. Review of literature and report of four cases.
In 1986, the Groupe d'Etude des Tumeurs osseuses (Bone Tumors Studying Group), organised a multicentric study of benign bone tumors and dystrophies localised in the hand. The result was very poor, because only five cases were found. Adding these cases to those already published in the literature, we report: 58 aneurysmal cyst, 12 chondromyxoid fibroma, 9 chondroblastoma, 6 bone cyst, 5 monostotic fibrous dysplasia. The total of 90 cases seems to confirm that the hand is a very unusual localisation for these tumors. These concerned equally metacarpal and phalanx for 37 each, but the carpus was concerned in only 7. Diagnosis was regularly missed before operation, and chondroma was usually suggested; the reason is probably that these tumors lost their usual characters in this particular localisation, except in two cases of fibrous dysplasia. In most cases curettage and cancellous graft seems sufficient to ascertain histology and cure the lesion.


Treatment of hypertension with chronotherapy: is it time of drug administration?
To review evidence for dosing antihypertensives at bedtime and possible cardiovascular risk reduction. A PubMed, EMBASE, and Cochrane Controlled Trials database literature search (1990-September 2014) limited to human subjects was performed using the search terms hypertension, chronotherapy, ambulatory blood pressure, morning administration, evening administration, and antihypertensives. Additional references were identified from literature citations. All prospective studies assessing cardiovascular outcomes or comparing morning to evening administration of antihypertensives were selected. Compared with morning administration, dosing one or more antihypertensive medications at bedtime helps induce a normal circadian blood pressure pattern and reduces the risk of cardiovascular disease morbidity and mortality in individuals with hypertension. Similar results have been reported in high-risk individuals with diabetes, chronic kidney disease, and resistant hypertension. A lack of diversity among studied populations and reliance on subgroup analyses are among the limitations of these data. All antihypertensive medications have not been studied in chronotherapy and do not uniformly achieve desired results. The most substantial evidence exists for medications affecting the renin-angiotensin-aldosterone system. Despite growing evidence and promise as a cost-effective strategy for reducing cardiovascular risk, chronotherapy is not uniformly recommended in the treatment of hypertension. Careful selection of patients and antihypertensives for chronotherapy is required. Further investigation is needed to evaluate the definitive impact of chronotherapy on cardiovascular outcomes.


Vitamin D status affects proteomic profile of HDL-associated proteins and inflammatory mediators in dyslipidemia.
Vitamin D deficiency and dyslipidemia have substantial implications for human health globally. Vitamin D is essential for bone metabolism and immune modulation, and its insufficiency is linked to various chronic inflammatory conditions. Dyslipidemia, characterized by low levels of high-density lipoprotein (HDL) and elevated levels of low-density lipoprotein (LDL) and triglycerides, is also prevalent. Previous research has shown a connection between vitamin D deficiency and low HDL, but the precise mechanism by which vitamin D influences HDL production and its anti-inflammatory properties remains unclear. This study aimed to investigate the proteomic profiles of individuals with and without vitamin D deficiency and dyslipidemia, specifically focusing on the effects of vitamin D on HDL production, its anti-inflammatory potential, and the molecular pathways associated with vitamin D deficiency and dyslipidemia, particularly inflammation and cancer pathways. By analyzing the proteomic profiles of 274 participants from the Qatar Biobank database, we identified 1301 proteins. Our findings indicated a decrease in HDL-associated apolipoproteins (ApoM and ApoD) in individuals with both dyslipidemia and vitamin D deficiency. Conversely, participants with these conditions exhibited increased expression of acute-phase proteins (SAA1 and SOD1), which are associated with inflammation. Pathway enrichment analysis revealed heightened inflammatory activity in individuals with vitamin D deficiency and dyslipidemia, with notable enrichments in pathways such as MAPK, JAK-STAT, Ras signaling, cytokine-cytokine receptor interaction, AGE-RAGE, ErbB signaling, and cancer pathways. Overall, cases of vitamin D deficiency showed enrichment in inflammation pathways, while individuals with both vitamin D deficiency and dyslipidemia demonstrated enhanced activation of cancer and inflammation pathways.


What schedule-induced polydipsia can tell us about alcoholism.
An animal model of chronic and excessive voluntary (unforced) alcohol ingestion is presented in which, by drinking, animals produce repeated, substantial elevations in blood ethanol concentration and develop physical dependence. The overindulgence is elective in that ethanol is chosen in preference to certain other fluid-ingestive alternatives. Beside the usual demonstrations of acutely compromised motor performance, tolerance development, cross-tolerance, etc., the model demonstrates that the consequences of even short, but continued, daily drinking episodes results in the disruption of reinforced behavior that occurs later in the day when blood ethanol is absent (impaired general functioning). The conditions which induce the ethanol overindulgence can generate a variety of behavioral excesses which places alcoholism in a context of environmentally determined malfunctions that are subject to therapeutic change by altering situational parameters. Efficacious experiments utilizing therapeutic and preventive strategies are described that may serve as suggestions for corresponding human alcoholism intervention strategies.


Haematological Parameters and Spleen Rate of Asymptomatic and Malaria Negative Children in Edo South District, Nigeria.
Malaria is commonly associated with alteration in haematologic cells of infected individuals in both the acute uncomplicated and severe phases. Whether this alteration occurs in the asymptomatic phase of the disease is still being investigated. To examine the haematocrit, thrombocytes, and monocytes levels of children with asymptomatic malaria compared with age/sex-matched controls who are malaria parasite negative and living in a stable malaria endemic region. It also set out to identify spleen rate of the children and to compare it with that observed in malaria negative controls. One hundred well-nourished children 2-9 years old with asymptomatic malaria parasitaemia and 100 age- and sex-matched malaria negative controls were recruited by multi-stage sampling from schools in a malaria endemic region of Nigeria. Malaria diagnosis was by microscopy, and each haematologic parameter was analysed following standard protocols. Mean (±) monocyte count of 2.25 ± 0.9 × 109 cells/L observed in asymptomatic malaria children was significantly higher than 1.34 ± 0.5 × 109 cells/L observed in those with no malaria (p = 0.00). Mean (±) thrombocyte count was significantly lower (asymptomatic 203.64 ± 45.90 × 109 cells/L Vs no malaria 230.91 ± 57.40 × 109 cells/L) (p = 0.00). Spleen rate in the children was 15.5%. Presence of splenomegaly was not statistically significantly fewer in children with asymptomatic malaria parasitaemia (ASMP) (14/31) when compared to those who were malaria parasite negative (17/31) (χ2 = 0.34, p = 0.57). Similarly, there was no significant difference in the mean [±] spleen length of children with ASMP (n = 14; 2.86 ± 0.9 cm) and those who were malaria negative (n = 17; 2.53 ± 0.6 cm) (t = 1.22, p = 0.23). Thrombocytopaenia and monocytosis could be pointers to malaria parasitaemia in asymptomatic phase in a stable malaria region.


Leachate generation from landfill in a semi-arid climate: A qualitative and quantitative study from Sousse, Tunisia.
Despite initiatives for enhanced recycling and waste utilization, landfill still represents the dominant disposal path for municipal solid waste (MSW). The environmental impacts of landfills depend on several factors, including waste composition, technical barriers, landfill operation and climatic conditions. A profound evaluation of all factors and their impact is necessary in order to evaluate the environmental hazards emanating from landfills. The present paper investigates a sanitary landfill located in a semi-arid climate (Tunisia) and highlights major differences in quantitative and qualitative leachate characteristics compared to landfills situated in moderate climates. Besides the qualitative analysis of leachate samples, a quantitative analysis including the simulation of leachate generation (using the HELP model) has been conducted. The results of the analysis indicate a high load of salts (Cl, Na, inorganic nitrogen) in the leachate compared to other landfills. Furthermore the simulations with HELP model highlight that a major part of the leachate generated originates form the water content of waste.


INFUSION OF CONVALESCENT PLASMA IS ASSOCIATED WITH CLINICAL IMPROVEMENT IN CRITICALLY ILL PATIENTS WITH COVID-19: A PILOT STUDY.
The ideal treatment of coronavirus disease (COVID)-19 has yet to be defined, but convalescent plasma (CoPla) has been successfully employed. The objective of the study was to study the safety and outcomes of the administration of CoPla to individuals with severe COVID-19 in an academic medical center. Ten patients were prospectively treated with plasma from COVID-19 convalescent donors. Over 8 days, the sequential organ failure assessment score dropped significantly in all patients, from 3 to 1.5 (p = 0.014); the Kirby index (PaO2/FiO2) score increased from 124 to 255, (p < 0.0001), body temperature decreased significantly from 38.1 to 36.9°C (p = 0.0058), and ferritin levels also dropped significantly from 1736.6 to 1061.8 ng/ml (p = 0.0001). Chest X-rays improved in 7/10 cases and in 6/10, computerized tomography scans also revealed improvement of the lung injury. Decreases in C-reactive protein and D-dimer levels were also observed. Three of five patients on mechanical ventilation support could be extubated, nine were transferred to conventional hospital floors, and six were sent home; two patients died. The administration of CoPla had no side effects and the 24-day overall survival was 77%. Although other treatments were also administered to the patients and as a result data are difficult to interpret, it seems that the addition of CoPla improved pulmonary function.


The structural basis for substrate promiscuity in 2-keto-3-deoxygluconate aldolase from the Entner-Doudoroff pathway in Sulfolobus solfataricus.
The hyperthermophilic Archaea Sulfolobus solfataricus grows optimally above 80 degrees C and metabolizes glucose by a non-phosphorylative variant of the Entner-Doudoroff pathway. In this pathway glucose dehydrogenase and gluconate dehydratase catalyze the oxidation of glucose to gluconate and the subsequent dehydration of gluconate to D-2-keto-3-deoxygluconate (KDG). KDG aldolase (KDGA) then catalyzes the cleavage of KDG to D-glyceraldehyde and pyruvate. It has recently been shown that all the enzymes of this pathway exhibit a catalytic promiscuity that also enables them to be used for the metabolism of galactose. This phenomenon, known as metabolic pathway promiscuity, depends crucially on the ability of KDGA to cleave KDG and D-2-keto-3-deoxygalactonate (KDGal), in both cases producing pyruvate and D-glyceraldehyde. In turn, the aldolase exhibits a remarkable lack of stereoselectivity in the condensation reaction of pyruvate and D-glyceraldehyde, forming a mixture of KDG and KDGal. We now report the structure of KDGA, determined by multiwavelength anomalous diffraction phasing, and confirm that it is a member of the tetrameric N-acetylneuraminate lyase superfamily of Schiff base-forming aldolases. Furthermore, by soaking crystals of the aldolase at more than 80 degrees C below its temperature activity optimum, we have been able to trap Schiff base complexes of the natural substrates pyruvate, KDG, KDGal, and pyruvate plus D-glyceraldehyde, which have allowed rationalization of the structural basis of promiscuous substrate recognition and catalysis. It is proposed that the active site of the enzyme is rigid to keep its thermostability but incorporates extra functionality to be promiscuous.


Thiazolidinediones in the treatment of type 2 diabetes.
In the last few years there has been an explosion of research that has improved our understanding of the pathogenesis of Type 2 diabetes mellitus (DM-2) and has led to the development of new oral antidiabetic drugs. Thiazolidinediones (TZDs) are the newest of these antidiabetic agents. TZDs are insulin sensitisers that depend on the presence of insulin for their action. They target insulin resistance, which is thought to play a central role in DM-2 and the associated metabolic syndrome characterised by central obesity, hypertension, dyslipidemia and hypercoagulability, all leading to increased cardiovascular morbidity and mortality. As a result, TZDs have the potential to improve other conditions associated with the metabolic syndrome, in addition to their glycaemic action. TZDs act by activating peroxisome proliferator-activated receptor (PPAR) phi a nuclear receptor implicated not only in lipid and glucose metabolism but other physiological functions as well. TZDs may have wide clinical applications beyond DM-2, as they can potentially be used to treat other conditions associated with insulin resistance and PPAR-phi receptors, such as impaired glucose tolerance, polycystic ovarian syndrome and HIV lipodystrophy.


Differential effects of culture senescence and mechanical stimulation on the proliferation and leiomyogenic differentiation of MSC from different sources: implications for engineering vascular grafts.
We examined the effects of senescence on the proliferation and leiomyogenic differentiation potential of mesenchymal stem cells (MSCs) isolated from bone marrow (BM-MSCs) or hair follicles (HF-MSCs). To this end, we compared ovine HF-MSCs and BM-MSCs in terms of their proliferation and differentiation potential to the smooth muscle cell lineage. We discovered that HF-MSCs are less susceptible to culture senescence compared with BM-MSCs. We hypothesized that application of mechanical forces may enhance the contractility and mechanical properties of vascular constructs prepared from senescent MSCs. Interestingly, HF-MSCs and BM-MSCs responded differently to changes in the mechanical microenvironment, suggesting that despite phenotypic similarities, MSCs from different anatomic locations may activate different pathways in response to the same microenvironmental factors. In turn, this may also suggest that cell-based tissue regeneration approaches may need to be tailored to the stem cell origin, donor age, and culture time for optimal results.


Typing of Vibrio vulnificus strains by variability in their 16S-23S rRNA intergenic spacer regions.
Amplification of the 16S-23S rDNA spacer region (ISR1) is a simple and rapid procedure for subtyping bacteria, especially those with several ribosomal operons including Vibrio vulnificus. V. vulnificus contains nine ribosomal operons with four or five ISR1 classes that differ in size and sequence. In the present study, 47 V. vulnificus strains of both shellfish and clinical origin were subtyped by their ISR1 patterns using "universal" primers, which target conserved sequences located in the 16S and the 23S rRNA genes. Sixteen different ISR1 patterns were observed that were grouped into two major clusters. Most (21/27, 77.8%) clinical isolates examined in this study grouped into a single cluster containing ISR1 patterns I, V, XI, and XII and these were highly similar (75%). This cluster was restricted to strains carrying the type B 16S rDNA (rrs) sequence which has been associated with human illness in previous studies. The remaining cluster consisted primarily of shellfish isolates. The highest variability in the ISR1 patterns was observed among shellfish isolates. Sequence analysis of the ISR1 region of selected strains demonstrated that all of them possess five ISR1 classes, with two "conserved sequence blocks" at the 5' and 3' end of the ISR1. All of these strains carried at least one tRNA gene and different classes differed in their tRNA gene composition. Some of the same ISR1 classes differed in size mainly due to an insertion of 35 bp in either of the conserved sequence blocks. These results demonstrate the feasibility of the ISR1 technique for V. vulnificus subtyping and suggest that ISR1 patterns appear to be linked to rrs sequence types and perhaps with virulence.


Differences in quality of life in rural and urban populations.
To examine the predictors of health related, quality of life in rural and urban populations. Parents and grandparents of students from 20 randomly selected primary schools in urban and rural areas of Kirikkale, Turkey were questioned for health-related quality of life (HRQL) and psychological distress, using the Short Form-12 (SF-12) Health Survey and Hospital Anxiety and Depression scale (HADS), respectively, which were returned by their children. Of 13,225 parents and grandparents 12,270 returned the questionnaires, for an overall response rate of 92.7%. SF-12 physical component summary (PCS), mental component summary (MCS), and overall scores were lower in participants from rural than those from urban areas. Mean HADS overall score was higher in subjects from rural than those from urban areas (16.6+/-6.8 vs. 14.8+/-6.8, P=0.0001). A linear regression model showed associations between SF-12 overall, PCS, and MCS scores with HAD total score after adjusting for sex, age, BMI, smoking, income, and education. Quality of life scores in subjects vary between areas. Psychological distress in subjects in rural areas may account for the poorer scores of quality of life in rural areas.


ZD1839: targeting the epidermal growth factor receptor in cancer therapy.
The erbB family of receptors, which includes the epidermal growth factor receptor, has been widely implicated in promoting proliferation of malignant cells. The critical role played by epidermal growth factor receptor in cancer has resulted in extensive research for selective inhibitors of the epidermal growth factor receptor signalling pathway. Selective small molecule epidermal growth factor receptor-tyrosine kinase inhibitors, such as ZD1839 (Iressa), block signal transduction pathways implicated in proliferation and survival of cancer cells and other host-dependent processes promoting cancer cell growth. In preclinical studies, ZD1839, alone and in combination with other agents, has demonstrated antitumour activity in a range of tumour types. Results from Phase I trials, in healthy volunteers and in patients with advanced disease, have shown that ZD1839 has excellent bioavailability and an acceptable tolerability profile. In these studies, ZD1839 has also shown promising clinical activity in patients with a variety of tumour types. Furthermore, Phase II studies confirmed clinically meaningful antitumour activity and have demonstrated symptom relief in the second- and third-line treatment of non-small cell lung cancer. Phase III trials are currently evaluating ZD1839 in combination with gemcitabine/cisplatin or paclitaxel/carboplatin as first-line treatment of non-small cell lung cancer and an ongoing clinical trial programme is investigating other tumours (i.e., head and neck, prostate, colon and breast) and other combinations. This article provides an overview of the current profile of ZD1839.


Reactive oxygen species modulate the barrier function of the human glomerular endothelial glycocalyx.
Reactive oxygen species (ROS) play a key role in the pathogenesis of proteinuria in glomerular diseases like diabetic nephropathy. Glomerular endothelial cell (GEnC) glycocalyx covers the luminal aspect of the glomerular capillary wall and makes an important contribution to the glomerular barrier. ROS are known to depolymerise glycosaminoglycan (GAG) chains of proteoglycans, which are crucial for the barrier function of GEnC glycocalyx. The aim of this study is to investigate the direct effects of ROS on the structure and function of GEnC glycocalyx using conditionally immortalised human GEnC. ROS were generated by exogenous hydrogen peroxide. Biosynthesis and cleavage of GAG chains was analyzed by radiolabelling (S(35) and (3)H-glucosamine). GAG chains were quantified on GEnC surface and in the cell supernatant using liquid chromatography and immunofluorescence techniques. Barrier properties were estimated by measuring trans-endothelial passage of albumin. ROS caused a significant loss of WGA lectin and heparan sulphate staining from the surface of GEnC. This lead to an increase in trans-endothelial albumin passage. The latter could be inhibited by catalase and superoxide dismutase. The effect of ROS on GEnC was not mediated via the GAG biosynthetic pathway. Quantification of radiolabelled GAG fractions in the supernatant confirmed that ROS directly caused shedding of HS GAG. This finding is clinically relevant and suggests a mechanism by which ROS may cause proteinuria in clinical conditions associated with high oxidative stress.