Structural characterization of novel oligosaccharides of cell-surface glycoproteins of Trypanosoma cruzi.
Affinity-purified glycopeptides were prepared from Trypanosoma cruzi using the carbohydrate-specific monoclonal antibody WIC29.26. These glycopeptides contain rhamnose, fucose, xylose, and galactose, in the ratio 1:1:2:3. A series of oligosaccharides was released from the glycopeptides by mild acid hydrolysis, while, in contrast, no oligosaccharides were released by either peptide N-glycosidase F or conventional base-catalyzed beta-elimination and reduction. This suggested the presence of a phosphodiester linkage between the carbohydrate and peptide, which was further supported by the detection of phosphothreonine in the glycopeptides. The mild acid liberated (MAL) fraction was resolved into two major acidic oligosaccharides (MAL-P1 and MAL-P2), two minor neutral oligosaccharides (MAL-P1b and MAL-P2b) and a neutral fraction (MAL-N1), consisting of Gal and Xyl monosaccharides. The MAL-P1 and MAL-P2 oligosaccharides proved to be hexa- and hepta-saccharides that shared a common xylose reducing terminus, but differed by one galactofuranose residue, and their negative charge was shown to be due to the presence of cyclic-phosphate attached to nonreducing terminal galactofuranose residues. The MAL-P1b and MAL-P2b oligosaccharides appeared to be nonphosphorylated versions of MAL-P1 and MAL-P2. Partial structures of MAL-P1 and MAL-P2 are suggested, based on compositional analyses, electrospray mass spectrometry, and tandem mass spectrometry before and after permethylation. The origin and significance of these unique trypanosomatid glycoconjugates is discussed.


Atelectasis and oxygenation in major surgery with either propofol with or without nitrous oxide or isoflurane anaesthesia.
Forty-two patients undergoing major colonic surgery were assigned at random to receive isoflurane-fentanyl anaesthesia with nitrous oxide in oxygen, propofol-fentanyl anaesthesia with air in oxygen or propofol-fentanyl anaesthesia with nitrous oxide in oxygen. The groups were comparable in demographic data. Atelectases were identified, and the area measured by computerised tomography of the chest 203 +/- 69 min after extubation, and oxygenation was determined by arterial blood gas samples taken during operation at 30, 60, 90 and 120 min after extubation and on postoperative days 1, 2 and 3. Atelectases were seen in all three groups with no differences in the mean area between groups. After operation, the effect of 4 l.min-1 of oxygen by nasal catheter on PaO2 was similar in all groups. A significant decrease in PaO2 was found during the first 3 days after surgery, and was also the same in all groups. There was no correlation between area of atelectasis and postoperative PaO2. We conclude there is no difference in the incidence of postoperative atelectasis or oxygenation when using propofol, with or without nitrous oxide or isoflurane.


Paraoxonase1-192 polymorphism modulates the nonfatal myocardial infarction risk associated with decreased HDLs.
Serum paraoxonase1 (PON1), a high density lipoprotein (HDL)-linked enzyme, appears to have a role in the protection of low density lipoproteins from oxidative stress. PON1 enzyme activity for paraoxon as a substrate is modulated, along with others at the PON1 locus, by the PON1-192 polymorphism, which contains low paraoxon-activity and high paraoxon-activity alleles (Q and R, respectively). The association of PON1 with HDL suggests that impaired serum concentrations of the lipoprotein could have consequences for the susceptibility to oxidative stress. Because PON1-192 polymorphism strongly influences PON1 activity toward paraoxon, we tested the hypothesis that this polymorphism may modulate the myocardial infarction (MI) risk associated with low HDL cholesterol concentrations. Two hundred eighty consecutive MI patients and 396 control subjects were studied. When considered as a whole, PON1-192 genetic polymorphism was not associated with higher MI risk. In the entire population, decreased HDL cholesterol concentration (<0.90 mmol/L in men and <1.11 mmol/L in women) conferred an MI risk of 2.56 (P=0.0001) compared with normal HDL levels. The risk increased to 4.51 (P<0.0001) in QQ homozygous HDL-deficient subjects relative to QQ homozygotes with normal HDL levels, decreased to 1.83 (P=0.1046) in QR heterozygote HDL-deficient subjects, and also decreased (to 1.41, P=0.6243) in RR homozygote HDL-deficient individuals compared with RR carriers with normal HDL cholesterol concentration. The effect of PON1-192 genotypes on the association of low HDL cholesterol levels and MI was related to gene dosage. A significantly decreased enzyme activity was found in HDL-deficient MI patients compared with HDL-deficient control subjects (median 208 U/L [interquartile range 136 to 298 U/L] versus median 235 U/L [interquartile range 163 to 350 U/L], respectively; P=0.025]. QQ homozygous MI patients showed the greatest difference of PON1 activity levels between normal and HDL-deficiency state groups (14.9%, P=0.002). Our observations raise the question of whether the decrease in PON1 activity and the MI risk associated with HDL deficiency are more evident in the low paraoxon-activity QQ genotype. It can be argued that the low paraoxon-activity QQ genotype, which may be adequate to prevent lipid peroxidation in normolipidemic subjects, may be insufficient when an HDL-deficiency state and low PON1 activity reflecting oxidative stress coexist. The risk of nonfatal MI is increased in HDL-deficiency states, principally among subjects carrying the low paraoxon-activity QQ genotype.


Fluorescence Blinking as an Output Signal for Biosensing.
We demonstrate the first biosensing strategy that relies on quantum dot (QD) fluorescence blinking to report the presence of a target molecule. Unlike other biosensors that utilize QDs, our method does not require the analyte to induce any fluorescence intensity or color changes, making it readily applicable to a wide range of target species. Instead, our approach relies on the understanding that blinking, a single particle phenomenon, is obscured when several QDs lie within the detection volume of a confocal microscope. If QDs are engineered to aggregate when they encounter a particular target molecule, the observation of quasi-continuous emission should indicate its presence. As proof of concept, we programmed DNAs to drive rapid isothermal assembly of QDs in the presence of a target strand (oncogene K-ras). The assemblies, confirmed by various gel techniques, contained multiple QDs and were readily distinguished from free QDs by the absence of blinking.


Dyadic Green's function of a cluster of spheres.
The electric dyadic Green's function (dGf) of a cluster of spheres is obtained by application of the superposition principle, dyadic algebra, and the indirect mode-matching method. The analysis results in a set of linear equations for the unknown, vector, wave amplitudes of the dGf; that set is solved by truncation and matrix inversion. The theory is exact in the sense that no simplifying assumptions are made in the analytical steps leading to the dGf, and it is general in the sense that any number, position, size and electrical properties can be considered for the spheres that cluster together. The point source can be anywhere, even within one of the spheres. Energy conservation, reciprocity, and other tests prove that this solution is correct. Numerical results are presented for an electric Hertz dipole radiating in the presence of an array of rexolite spheres, which manifests lensing and beam-forming capabilities.


Effect of intracardiac repair on biosynthesis of thromboxane A2 and prostacyclin in children with a left to right shunt.
To investigate the effect of intracardiac repair on the abnormal biosynthesis of prostacyclin (PGI2) and thromboxane A2 (TXA2) in children with congenital heart disease and increased pulmonary blood flow. A prospective study with immunoaffinity chromatography and gas chromatography-mass spectrometry to measure the urinary excretion products of PGI2 (2,3-dinor-6-oxo-prostaglandin (PG) F1 alpha (2,3-dinor-6-oxo-PGF1 alpha)) and TXA2 (2,3-dinor-TXB2) before operation, in the first 12-24 h after operation, and at discharge from hospital. A supraregional referral centre for patients with congenital heart disease. 15 patients aged 2 to 60 months (median 7 months) with a left to right shunt who underwent intracardiac repair. The preoperative 2,3-dinor-TXB2 excretion rate was greater than that found previously in a control group of 16 healthy children with a median (range) age of 24 (6-36) months (1159(201) v 592(122) ng/g creatinine in controls, P = 0.006). The excretion rate rose after operation to 9600(3832) ng/g creatinine (P = 0.01) and decreased before discharge to 1071(191) ng/g creatinine (NS), but remained greater than that of the control group (P = 0.014). Before operation 2,3-dinor-6-oxo-PGF1 alpha excretion rates were similar to those of the healthy children (482(68) v 589(95) ng/g creatinine in controls) but increased after operation to 19,668(11,162) ng/creatinine (P = 0.002) and fell at discharge to 1621(245) ng/g creatinine although this was higher than both preoperative and control rates (P = 0.005 and P = 0.0002 respectively). The preoperative ratio of 2,3-dinor-TXB2 to 2,3-dinor-6-oxo-PGF1 alpha excretion was greater than that of the control group (3.2(0.8) v 1.3(0.22) in controls, (P = 0.005)), decreased significantly after operation to 0.9(0.13) (P = 0.016), and changed little, to 0.7(0.12), before discharge. The last two ratios were similar to those in normal children and significantly lower than those before operation (P = 0.004). In children with a left to right shunt the ratio of the excretion rates of the metabolites of TXA2 and PGI2 was abnormal before operation, which favoured vasoconstriction and platelet aggregation, but had decreased at discharge from hospital. The increase in excretion of PGI2 metabolites over TXA2 metabolite after intracardiac repair augurs well for pulmonary vascular recovery.


Temporal Changes in Skeletal Muscle Capillary Responses and Endothelial-Derived Vasodilators in Obesity-Related Insulin Resistance.
The inability of insulin to increase skeletal muscle capillary blood volume (CBV) reduces glucose uptake in insulin resistance (IR). We hypothesized that abnormalities in endothelial-derived vasodilator pathways are temporally associated with the development of IR and an impaired ability to increase skeletal muscle CBV. A comprehensive metabolic and vascular screening assessment was performed on 10 adult rhesus macaques at baseline and every 4-6 months for 2 years after starting a high-fat diet supplemented with fructose. Diet changes resulted in an 80% increase in truncal fat by 4 months. Hyperinsulinemia and decreased glucose utilization were observed from 4 to 18 months. At 24 months, pancreatic secretory function and the glucose utilization rate declined. CBV at rest and during an intravenous glucose tolerance test demonstrated a sustained increase from 4 to 18 months and then abruptly fell at 24 months. Nitric oxide bioavailability progressively decreased over 2 years. Conversely, endothelial-derived vasodilators progressively increased over 18 months and then abruptly decreased at 24 months in concert with the CBV. The increase in basal and glucose-mediated CBV early in IR may represent a compensatory response through endothelial-derived vasodilator pathways. The inability to sustain a vascular compensatory response limits glucose-mediated increases in CBV, which correlates with the severity of IR.


Quantifying calf mortality on dairy farms: Challenges and solutions.
In the Netherlands, the mortality rate of ear-tagged calves <1 yr is one of the indicators that is continuously monitored in census data and is defined as the number of deceased calves relative to the number of calf-days-at-risk. In 2017, yearly calf mortality rates were published in the lay press and resulted in discussions about the calculation of this parameter among stakeholders because the same parameter appeared to be calculated in many different ways by different organizations. These diverse definitions of calf mortality answered different aims such as early detection of deviations, monitoring trends, or providing insight into herd-specific results, but were difficult to understand by stakeholders. The aim of this study was to evaluate several definitions of calf mortality for scientific validity, usefulness for policymakers, and comprehensibility by farmers. Based on expert consultations, 10 definitions for calf mortality were evaluated that assessed different age categories, time periods, and denominators. Differences in definitions appeared to have a large effect on the magnitude of mortality. For example, with the original mortality parameter, the mortality rate was 16.5% per year. When the first year of life was subdivided into 3 age categories, the mortality rate was 3.3, 4.5, and 3.1% for postnatal calves (≤14 d), preweaned calves (15-55 d), and weaned calves (56 d-1 yr), respectively. Although it was logical that these mortality rates were lower than the original, the sum of the 3 separate mortality rates was also lower than the original mortality rate. The reason was that the number of calves present in a herd and the risk of mortality are not randomly distributed over a calf's first year of life and the conditional nature of mortality rates when calculated for different age categories. Ultimately, 4 parameters to monitor calf mortality in Dutch dairy herds were chosen based on scientific value, usefulness for monitoring of trends, and comprehensibility by farmers: perinatal calf mortality risk (i.e., mortality before, during, or shortly after the moment of birth up to the moment of ear-tagging), postnatal calf mortality risk (≤14 d), preweaned calf mortality rate (15-55 d), and weaned calf mortality rate (56 d-1 yr). Slight differences in definitions of parameters can have a major effect on results, and many factors have to be taken into account when defining an important health indicator such as mortality. Our evaluation resulted in a more thorough understanding of the definitions of the selected parameters and agreement by the stakeholders to use these key indicators to monitor calf mortality.


Behavioral consequences of bone marrow transplantation in the treatment of murine mucopolysaccharidosis type VII.
The gusmps/gusmps mouse is a model of the human lysosomal storage disease mucopolysaccharidosis type VII caused by deficient beta-glucuronidase activity. Bone marrow transplantation has been shown to correct some of their biochemical and pathological abnormalities but its efficacy in correcting their neurological functional deficits is unknown. We transplanted the neonatal gusmps/gusmps mice and their normal controls and evaluated their central nervous system function with two behavioral tests: the grooming test, a developmentally regulated and genetically based activity, and a Morris water maze test which assessed spatial learning abilities. The two transplanted groups groomed less than the normals, were unable to remember the location of an invisible platform from day to day, and were severely impaired at developing strategies to locate the platform in unfamiliar locations. The performance of both normal and mutant transplanted groups was clearly inferior to the untreated normals and, in some instances, close to or worse than the untreated mutants, even though the enzyme abnormalities of the mutants have been partially corrected. Hence, the behavioral deficits in the mutant mice were not restored to normal while similarly treated normal mice showed significant functional deterioration, indicating the detrimental consequence of this therapy in the neonatal period.


Evaluation of the immunocrit method to detect failure of passively acquired immunity in dairy calves.
OBJECTIVE To evaluate the usefulness of serum immunocrit measurement to detect failure of passively acquired immunity (FPI) in dairy calves. DESIGN Diagnostic test evaluation. ANIMALS 249 female dairy calves (age, 2 to 6 days). PROCEDURES A blood sample was collected from each calf, and serum was harvested. Immunocrit was measured in serum samples by use of 55% ammonium sulfate solution and the standard technique. Serum IgG concentration was measured by means of radial immunodiffusion (reference standard), with FPI defined as a result < 1,000 mg/dL. The immunocrit value (cutpoint) that maximized both sensitivity and specificity of the method for detection of FPI was determined by construction of receiver operating characteristic curves, and likelihood ratios for positive and negative test results were calculated. RESULTS Immunocrit values were significantly correlated (ρ = 0.71) with serum IgG concentration as measured by radial immunodiffusion. An immunocrit cutpoint of 11% was optimal for detection of FPI in the calves. Sensitivity and specificity of the immunocrit method at this cutpoint were 0.88 (95% confidence interval [CI], 0.79 to 0.97) and 0.51 (95% CI, 0.44 to 0.58), respectively. Likelihood ratios for positive and negative test results were 1.80 (95% CI, 1.51 to 2.14) and 0.23 (95% CI, 0.11 to 0.51), respectively. CONCLUSIONS AND CLINICAL RELEVANCE The immunocrit method was useful for identifying dairy calves with FPI and was simple and could be quickly performed. Because precipitation of immunoglobulins by ammonium sulfate is not species specific, the immunocrit method should be evaluated for detection of FPI in other veterinary species as well.


Sost deficiency leads to reduced mechanical strains at the tibia midshaft in strain-matched in vivo loading experiments in mice.
Sclerostin, a product of the Sost gene, is a Wnt-inhibitor and thus negatively regulates bone accrual. Canonical Wnt/β-catenin signalling is also known to be activated in mechanotransduction. Sclerostin neutralizing antibodies are being tested in ongoing clinical trials to target osteoporosis and osteogenesis imperfecta but their interaction with mechanical stimuli on bone formation remains unclear. Sost knockout (KO) mice were examined to gain insight into how long-term Sost deficiency alters the local mechanical environment within the bone. This knowledge is crucial as the strain environment regulates bone adaptation. We characterized the bone geometry at the tibial midshaft of young and adult Sost KO and age-matched littermate control (LC) mice using microcomputed tomography imaging. The cortical area and the minimal and maximal moment of inertia were higher in Sost KO than in LC mice, whereas no difference was detected in either the anterior-posterior or medio-lateral bone curvature. Differences observed between age-matched genotypes were greater in adult mice. We analysed the local mechanical environment in the bone using finite-element models (FEMs), which showed that strains in the tibiae of Sost KO mice are lower than in age-matched LC mice at the diaphyseal midshaft, a region commonly used to assess cortical bone formation and resorption. Our FEMs also suggested that tissue mineral density is only a minor contributor to the strain distribution in tibial cortical bone from Sost KO mice compared to bone geometry. Furthermore, they indicated that although strain gauging experiments matched strains at the gauge site, strains along the tibial length were not comparable between age-matched Sost KO and LC mice or between young and adult animals within the same genotype.


Unsettling moods in rural midwifery practice.
Rural midwifery and maternity care is vulnerable due to geographical isolation, staffing recruitment and retention. Highlighting the concerns within rural midwifery is important for safe sustainable service delivery. Hermeneutic phenomenological study undertaken in New Zealand (NZ). 13 participants were recruited in rural regions through snowball technique and interviewed. Transcribed interview data was interpretively analysed. Findings are discussed through the use of philosophical notions and related published literature. Unsettling mood of anxiety was revealed in two themes (a) 'Moments of rural practice' as panicky moments; an emergency moment; the unexpected moment and (b) 'Feelings of being judged' as fearing criticism; fear of the unexpected happening to 'me' fear of losing my reputation; fear of feeling blamed; fear of being identified. Although the reality of rural maternity can be more challenging due to geographic location than urban areas this need not be a reason to further isolate these communities through negative judgement and decontextualized policy. Fear of what was happening now and something possibly happening in the future were part of the midwives' reality. The joy and delight of working rurally can become overshadowed by a tide of unsettling and disempowering fears. Positive images of rural midwifery need dissemination. It is essential that rural midwives and their communities are heard at all levels if their vulnerability is to be lessened and sustainable safe rural communities strengthened.


Envelope and pre-membrane protein structural amino acid mutations mediate diminished avian growth and virulence of a Mexican West Nile virus isolate.
The hallmark attribute of North American West Nile virus (WNV) strains has been high pathogenicity in certain bird species. Surprisingly, this avian virulent WNV phenotype has not been observed during its geographical expansion into the Caribbean, Central America and South America. One WNV variant (TM171-03-pp1) isolated in Mexico has demonstrated an attenuated phenotype in two widely distributed North American bird species, American crows (AMCRs) and house sparrows (HOSPs). In order to identify genetic determinants associated with attenuated avian replication of the TM171-03-pp1 variant, chimeric viruses between the NY99 and Mexican strains were generated, and their replicative capacity was assessed in cell culture and in AMCR, HOSP and house finch avian hosts. The results demonstrated that mutations in both the pre-membrane (prM-I141T) and envelope (E-S156P) genes mediated the attenuation phenotype of the WNV TM171-03-pp1 variant in a chicken macrophage cell line and in all three avian species assayed. Inclusion of the prM-I141T and E-S156P TM171-03-pp1 mutations in the NY99 backbone was necessary to achieve the avian attenuation level of the Mexican virus. Furthermore, reciprocal incorporation of both prM-T141I and E-P156S substitutions into the Mexican virus genome was necessary to generate a virus that exhibited avian virulence equivalent to the NY99 virus. These structural changes may indicate the presence of new evolutionary pressures exerted on WNV populations circulating in Latin America or may signify a genetic bottleneck that has constrained their epiornitic potential in alternative geographical locations.


In vitro and in vivo antibacterial activities of the glycylcyclines, a new class of semisynthetic tetracyclines.
N,N-Dimethylglycylamido (DMG) derivatives of minocycline and 6-demethyl-6-deoxytetracycline are new semisynthetic tetracyclines referred to as the glycylcyclines. The in vitro activities of the glycylcyclines were evaluated in comparison with those of minocycline and tetracycline against strains carrying characterized tetracycline resistance determinants and against 995 recent clinical isolates obtained from geographically distinct medical centers in North America. The glycylcyclines were active against tetracycline-resistant strains carrying efflux [tet(A), tet(B), tet(C), and tet(D) in Escherichia coli and tet(K) in Staphylococcus aureus] and ribosomal protection [tet(M) in S. aureus, Enterococcus faecalis, and E. coli)] resistance determinants. Potent activity (MIC for 90% of strains, < or = 0.5 microgram/ml) was obtained with the glycylcyclines against methicillin-susceptible and methicillin-resistant S. aureus, E. faecalis, Enterococcus faecium, and various streptococcal species. The glycylcyclines exhibited good activity against a wide diversity of gram-negative aerobic and anaerobic bacteria, most of which were less susceptible to minocycline and tetracycline. The activities of the glycylcyclines against most organisms tested were comparable to each other. The in vivo efficacies of the glycylcyclines against acute lethal infections in mice when dosed intravenously were reflective of their in vitro activities. The glycylcyclines had efficacies comparable to that of minocycline against infections with methicillin-susceptible and methicillin-resistant S. aureus strains, a strain carrying tet(K), and a tetracycline-susceptible E. coli strain but exceeded the effectiveness of minocycline against infections with resistant isolates, including strains harboring tet(M) or tet(B). Levels of DMG-6-deoxytetracycline in serum were higher and more sustained than those of DMG-minocycline or minocycline. Our results show that the glycylcyclines have potent in vitro activities against a wide spectrum of gram-positive and gram-negative, aerobic and anaerobic bacteria, including many resistant strains. On the basis of their in vitro and in vivo activities, the glycylcyclines represent a significant advance to the tetracycline class of antibiotics and have good potential value for clinical efficacy.


Study of the E2 gene product of the cottontail rabbit papillomavirus reveals a common mechanism of transactivation among papillomaviruses.
The long control region (LCR) of the cottontail rabbit papillomavirus (CRPV) harbors a transcriptional promoter which can be transactivated, as reflected by cat gene expression, by cotransfection with plasmids which express the intact E2 open reading frame of CRPV, human papillomavirus type 18 (HPV18), and bovine papillomavirus type 1 (BPV1). The E2 protein of CRPV can also transactivate the LCRs of BPV1, HPV1, and HPV18 inserted in front of the cat gene in enhancer or promoter configuration. Competition experiments in vivo and binding studies with CRPV E2 protein synthesized in vitro suggest that the different E2 proteins transactivate transcription by a common mechanism involving binding to the same ACCG-CGGT target sequence. The C-terminal part of the protein is necessary for its DNA-binding function. Analysis of the transactivation data and of the LCR sequences of these four viruses suggests that the two cutaneous viruses (CRPV and BPV1) present a similar pattern of promoter regulation but that the activity of the promoters of genital human viruses is less dependent on E2 regulation and is at least partially regulated by cellular factors.


The Association Between Mutuality, Anxiety, and Depression in Heart Failure Patient-Caregiver Dyads: An Actor-Partner Interdependence Model Analysis.
Heart failure (HF) patient-caregiver dyads experience severe psychological problems, such as anxiety and depression. A variable that has been found to be associated with anxiety and depression in patients and caregivers in severe chronic conditions is mutuality. However, this association has not been explored in HF patient-caregiver dyads to date. The aim of this study was to evaluate the associations among mutuality, anxiety, and depression in HF patient-caregiver dyads. This was a cross-sectional study. Mutuality, anxiety, and depression in HF patient-caregiver dyads were assessed using the Mutuality Scale (MS) total and 4 dimension scores and the Hospital Anxiety and Depression Scale, respectively. Data were analyzed using the actor-partner interdependence model to examine how mutuality of patients and caregivers was associated with both the patients' own (actor effect) and their partners' anxiety and depression (partner effect). A sample of 366 dyads of patients with HF (mean age, 72 years; 56% male) and caregivers (mean age, 54 years; 73.3% female) was enrolled. Regarding patient anxiety, we observed only an actor effect between the MS dimension scores of "love and affection" and "reciprocity" and anxiety in patients (B = -1.108, P = .004 and B = -0.826, P = .029, respectively). No actor and partner effects were observed concerning caregiver anxiety. Regarding depression, we observed that only the MS dimension of "love and affection" in patients had both an actor (patient: B = -0.717, P = .032) and a partner (caregiver: B = 0.710, P = .040) effect on depression. The assessment of MS in HF patient-caregiver dyads is important to formulate interventions aimed at improving anxiety and depression in patients and caregivers.


Isolation, characterization and gene sequence analysis of a membrane-associated 89 kDa Fe(III) reducing cytochrome c from Geobacter sulfurreducens.
Geobacter sulfurreducens is capable of anaerobic respiration with Fe(III) as a terminal electron acceptor via a membrane-bound Fe(III) reductase activity associated with a large molecular mass cytochrome c. This cytochrome was purified by detergent extraction of the membrane fraction, Q-Sepharose ion-exchange chromatography, preparative electrophoresis, and MonoQ ion-exchange chromatography. Spectrophotometric analysis of the purified cytochrome reveals a c-type haem, with no evidence of haem a, haem b or sirohaem. The cytochrome has an M(r) of 89000 as determined by denaturing PAGE, and has an isoelectric point of 5.2 as determined by analytical isoelectric focusing. Dithionite-reduced cytochrome can donate electrons to Fe(III)-nitrilotriacetic acid and synthetic ferrihydrite, thus demonstrating that the cytochrome has redox and thermodynamic properties required for reduction of Fe(III). Analysis using cyclic voltammetry confirmed that the reduced cytochrome can catalytically transfer electrons to ferrihydrite, further demonstrating its ability to be an electron transport mediator in anaerobic Fe(III) respiration. Sequence analysis of a cloned chromosomal DNA fragment revealed a 2307 bp open reading frame (ferA) encoding a 768 amino acid protein corresponding to the 89 kDa cytochrome. The deduced amino acid sequence (FerA) translated from the open reading frame contained 12 putative haem-binding motifs, as well as a hydrophobic N-terminal membrane anchor sequence, a lipid-attachment site and an ATP/GTP-binding site. FerA displayed 20% or less identity with amino acid sequences of other known cytochromes, although it does share some features with characterized polyhaem cytochromes c.


Prevention of postoperative abdominal adhesions by a novel, glycerol/sodium hyaluronate/carboxymethylcellulose-based bioresorbable membrane: a prospective, randomized, evaluator-blinded multicenter study.
Postoperative abdominal adhesions are associated with significant morbidity and mortality, placing a substantial burden on healthcare systems worldwide. Development of a bioresorbable membrane containing up to 23 percent glycerol and chemically modified sodium hyaluronate/carboxymethylcellulose offers ease of handling and has been shown to provide significant postoperative adhesion prevention in animals. This study was designed to assess the safety of glycerol hyaluronate/carboxymethylcellulose and to evaluate its efficacy in reducing the incidence, extent, and severity of postoperative adhesion development in surgical patients. Twelve centers enrolled 120 patients with ulcerative colitis or familial polyposis who were scheduled for a restorative proctocolectomy and ileal pouch-anal anastomosis with diverting loop ileostomy. Before surgical closure, patients were randomized to no anti-adhesion treatment (control) or treatment with glycerol hyaluronate/carboxymethylcellulose membrane under the midline incision. At ileostomy closure, laparoscopy was used to evaluate the incidence, extent, and severity of adhesion formation to the midline incision. Data were analyzed using the intent-to-treat population. Treatment with glycerol hyaluronate/carboxymethylcellulose resulted in 19 of 58 patients (33 percent) with no adhesions compared with 6 of 60 adhesion-free patients (10 percent) in the no treatment control group (P = 0.002). The mean extent of postoperative adhesions to the midline incision was significantly lower among patients treated with glycerol hyaluronate/carboxymethylcellulose compared with patients in the control group (P < 0.001). The severity of postoperative adhesions to the midline incision was significantly less with glycerol hyaluronate/carboxymethylcellulose than with control (P < 0.001). Adverse events were similar between treatment and no treatment control groups with the exception of abscess and incisional wound complications were more frequently observed with glycerol hyaluronate/carboxymethylcellulose. Glycerol hyaluronate/carboxymethylcellulose was shown to effectively reduce adhesions to the midline incision and adhesions between the omentum and small bowel after abdominal surgery. Safety profiles for the treatment and no treatment control groups were similar with the exception of more infection complications associated with glycerol hyaluronate/carboxymethylcellulose use. Animal models did not predict these complications.


Larval size and age affect colonization in a marine invertebrate.
The relationship between offspring size and performance determines the optimal trade-off between producing many small offspring or fewer large offspring and the existence of this relationship has become a central tenet of life-history theory. For organisms with multiple life-history stages, the relationship between offspring size and performance is determined by the effects of offspring size in each life-history stage. Marine invertebrates have long been a model system for examining the evolutionary ecology of offspring size, and whilst offspring size effects have been found in several life-history stages, the crucial stage of colonization has received less attention. We examined the effect of offspring size on the settlement response of sea-urchin larvae (Heliocidaris erythrogramma) to preferred and less preferred host plants, how these effects changed over the larval period and estimated the success of juveniles in the field on preferred and less-preferred host plants. We found that smaller larvae became competent to respond to preferred host plant cues sooner than larger larvae but larger larvae rejected less-preferred host plants for longer than smaller larvae. Overall, smaller H. erythrogramma larvae are likely to have less dispersal potential and are more likely to settle in less-preferred habitats whereas larger larvae appear to have an obligately longer dispersal period but settle in preferred habitats. Our results suggest that marine invertebrates that produce non-feeding larvae may have the potential to affect the dispersal of their offspring in previously unanticipated ways and that offspring size is subject to a complex web of selection across life-history stages.


Genetic and environmental influences on dentofacial structures and oral health: studies of Australian twins and their families.
Our studies of the teeth and faces of Australian twins commenced at the School of Dentistry, The University of Adelaide in the early 1980s. There are now over 900 pairs of twins enrolled in our continuing investigations, together with 1200 relatives. There are 3 main cohorts of participants. The first cohort comprises around 300 pairs of teenage twins for whom various records have been collected, including dental casts, facial photographs, finger and palm prints and information on laterality, including handedness. The second cohort comprises around 300 pairs of twins who have been examined at 3 stages of dental development from approximately 4 years of age to about 14 years: at primary, mixed, and permanent dentition (excluding 3rd molars) stages. The most recent study of tooth emergence and oral health, for which we are currently recruiting twins, will provide a third cohort of around 500 twin pairs aged from around birth to 3 to 4 years of age. Our broad aim in these studies has been to improve our understanding of how genetic and environmental factors contribute to variation in dental and facial features, and to oral health. We have also used our data to investigate aspects of the determination of laterality, particularly the fascinating phenomenon of mirror imaging. We plan to maximize the use of the longitudinal data and DNA we have collected, and continue to collect, by performing genome-wide scans for putative genetic linkage peaks for a range of dental features, and then to test for association between a series of likely candidate genes and our phenotypes.


Dynamic shapes of the zygote and two-cell mouse and human.
Mouse zygote morphokinetics were measured during interphase, the mitotic period, cytokinesis, and two-cell stage. Sequences of rounder-distorted-rounder shapes were revealed, as were changing patterns of cross section area. A calcium chelator and an actin-disrupting agent inhibited the area changes that occurred between pronuclear envelope breakdown and cytokinesis. During cell division, two vortices developed in each nascent cell and they rotated in opposite directions at each end of the cell, a pattern that sometimes persisted for up to 10 h. Exchange with the environment may have been promoted by these shape and area cycles and persisting circulation in the cytoplasm may have a similar function between a cell's interior and periphery. Some of these movements were sporadically also seen in human zygotes with abnormal numbers of pronuclei and the two-cell stages that developed from these compromised human zygotes.


Admission to hospital of singleton children born following assisted reproductive technology (ART).
Adverse perinatal outcomes are more common in singletons born following assisted reproductive technology (ART) and this would predict an increase in hospitalization during infancy and early childhood. We investigated hospital admissions during the first 3 years of life for all singleton children born in Western Australia between 1994 and 2000 [1328 ART, 162 350 spontaneously conceived (SC)]. ART infants had a significantly longer birth admission and were four times more likely to be admitted to neonatal intensive care units (NICU) than SC infants. ART children had a 60% greater risk of one or more admissions in their first year and an equal risk of admission in their second and third years. Their length of stay in hospital was longer in each age period. Maternal, infant and socio-economic confounders accounted for most of the increased admission risk in the first year. However, after adjustment, a 20% increase in the risk of admission to NICU (P < 0.05) and admission to hospital during the first year (P < 0.05) remained. Couples undertaking ART should be aware that ART infants are more likely to be admitted to a NICU, to be hospitalized in the first year of life and to stay in hospital longer than other children.


Monoclonal antibodies specific for interleukin 3-sensitive murine cells.
The mAb CC11 and CB5 reacted against all 18 IL-3-dependent cell lines tested, but not against cells insensitive to IL-3. Up to 53% nucleated cells from fetal liver (14th day of gestation) and 79% bone marrow cells of young adult mice were positive for both CC11 and CB5 antigens, but cells from thymus, lymph node, heart, and kidney were negative. The molecule recognized by both antibodies has an Mr of 50,000-70,000, a pI of 5.7-6.2, and carries heterogeneous N-linked glycans of high Mr. Both CC11 and CB5 specifically inhibited the growth of clones supported by rIL-3. Neither antibody affected the action of IL-1, IL-2, or B cell maturation factor; the proliferative responses of splenocytes to Con A, PWM, and LPS; nor the maturation of spleen B cells into antibody-secreting cells stimulated by LPS. rIL-3 specifically modulated the expression of the CC11/CB5 glycoprotein on the cell membrane of IL-3-dependent clones. Finally, freshly isolated bone marrow cells that have the CC11/CB5 glycoprotein on the cell membrane proliferated in response to IL-3, whereas cells that lack this molecule did not. We suggest that CC11 and CB5 react against receptors for mouse IL-3.


Executive control of attention in narcolepsy.
Narcolepsy with cataplexy (NC) is a disabling sleep disorder characterized by early loss of hypocretin neurons that project to areas involved in the attention network. We characterized the executive control of attention in drug-free patients with NC to determine whether the executive deficits observed in patients with NC are specific to the disease itself or whether they reflect performance changes due to the severity of excessive daytime sleepiness. Twenty-two patients with NC compared to 22 patients with narcolepsy without cataplexy (NwC) matched for age, gender, intellectual level, objective daytime sleepiness and number of sleep onset REM periods (SOREMPs) were studied. Thirty-two matched healthy controls were included. All participants underwent a standardized interview, completed questionnaires, and neuropsychological tests. All patients underwent a polysomnography followed by multiple sleep latency tests (MSLT), with neuropsychological evaluation performed the same day between MSLT sessions. Irrespective of diagnosis, patients reported higher self-reported attentional complaints associated with the intensity of depressive symptoms. Patients with NC performed slower and more variably on simple reaction time tasks than patients with NwC, who did not differ from controls. Patients with NC and NwC generally performed slower, reacted more variably, and made more errors than controls on executive functioning tests. Individual profile analyses showed a clear heterogeneity of the severity of executive deficit. This severity was related to objective sleepiness, higher number of SOREMPs on the MSLT, and lower intelligence quotient. The nature and severity of the executive deficits were unrelated to NC and NwC diagnosis. We demonstrated that drug-free patients with NC and NwC complained of attention deficit, with altered executive control of attention being explained by the severity of objective sleepiness and global intellectual level. Further studies are needed to explore whether medications that promote wakefulness can improve the executive functions in narcolepsy.


Isolation, purification, and identification of an important pigment, sepiapterin, from integument of the lemon mutant of the silkworm, Bombyx mori.
Sepiapterin is the precursor of tetrahydrobiopterin, an important coenzyme of aromatic amino acid hydroxylases, the lack of which leads to a variety of physiological metabolic diseases or neurological syndromes in humans. Sepiapterin is a main pigment component in the integument of the lemon mutant of the silkworm, Bombyx mori (L.) (Lepidoptera: Bombycidae), and is present there in extremely high content, so lemon is a valuable genetic resource to extract sepiapterin. In this study, an effective experimental system was set up for isolation and purification of sepiapterin from lemon silkworms by optimizing homogenization solvent, elution buffer, and separation chromatographic column. The results showed that ethanol was the most suitable solvent to homogenize the integument, with a concentration of 50% and solid:liquid ratio of 1:20 (g/mL). Sepiapterin was purified successively by column chromatography of cellulose Ecteola, sephadex G-25-150, and cellulose phosphate, and was identified by ultraviolet-visible absorption spectrometry. A stable and accurate high performance liquid chromatography method was constructed to identify sepiapterin and conduct qualitative and quantitative analyses. Sepiapterin of high purity was achieved, and the harvest reached about 40 ug/g of integument in the experiments. This work helps to obtaining natural sepiapterin in large amounts in order to use the lemon B. mori mutant to produce BH4 in vitro.


Extended prophylaxis of venous thromboembolism with idraparinux.
The extended use of vitamin K antagonists for prophylaxis against venous thromboembolism is often constrained by risk-benefit limitations and inconvenience. We evaluated the efficacy and safety of a 6-month extension of prophylaxis against recurrent venous thromboembolism with idraparinux in patients who had initially received 6 months of prophylaxis with an anticoagulant. We randomly assigned patients who had completed 6 months of prophylaxis with idraparinux or a vitamin K antagonist and in whom extended anticoagulation was warranted to receive once-weekly injections of 2.5 mg of idraparinux or placebo for 6 months without monitoring. The primary efficacy and safety outcomes were recurrent venous thromboembolism and major bleeding. Of 1215 patients, 6 of 594 (1.0%) in the idraparinux group and 23 of 621 (3.7%) in the placebo group had recurrent venous thromboembolism (P=0.002). Major bleeding occurred in 11 patients (1.9%) in the idraparinux group and in none in the placebo group (P<0.001). Of these 11 episodes, 3 were fatal intracranial hemorrhages. As compared with patients whose initial treatment was a vitamin K antagonist, patients whose initial treatment was idraparinux who were assigned to 6 months in the placebo group had a lower incidence of recurrent thromboembolism (0.7% vs. 5.9%); patients who received 6 additional months of idraparinux therapy had a higher incidence of major bleeding (3.1% vs. 0.9%). During a 6-month extension of thromboprophylaxis, idraparinux was effective in preventing recurrent thromboembolism but was associated with an increased risk of a major hemorrhage. (ClinicalTrials.gov number, NCT00071279 [ClinicalTrials.gov].).


The Role of Datasets on Scientific Influence within Conflict Research.
We inductively tested if a coherent field of inquiry in human conflict research emerged in an analysis of published research involving "conflict" in the Web of Science (WoS) over a 66-year period (1945-2011). We created a citation network that linked the 62,504 WoS records and their cited literature. We performed a critical path analysis (CPA), a specialized social network analysis on this citation network (~1.5 million works), to highlight the main contributions in conflict research and to test if research on conflict has in fact evolved to represent a coherent field of inquiry. Out of this vast dataset, 49 academic works were highlighted by the CPA suggesting a coherent field of inquiry; which means that researchers in the field acknowledge seminal contributions and share a common knowledge base. Other conflict concepts that were also analyzed-such as interpersonal conflict or conflict among pharmaceuticals, for example, did not form their own CP. A single path formed, meaning that there was a cohesive set of ideas that built upon previous research. This is in contrast to a main path analysis of conflict from 1957-1971 where ideas didn't persist in that multiple paths existed and died or emerged reflecting lack of scientific coherence (Carley, Hummon, and Harty, 1993). The critical path consisted of a number of key features: 1) Concepts that built throughout include the notion that resource availability drives conflict, which emerged in the 1960s-1990s and continued on until 2011. More recent intrastate studies that focused on inequalities emerged from interstate studies on the democracy of peace earlier on the path. 2) Recent research on the path focused on forecasting conflict, which depends on well-developed metrics and theories to model. 3) We used keyword analysis to independently show how the CP was topically linked (i.e., through democracy, modeling, resources, and geography). Publically available conflict datasets developed early on helped shape the operationalization of conflict. In fact, 94% of the works on the CP that analyzed data either relied on publically available datasets, or they generated a dataset and made it public. These datasets appear to be important in the development of conflict research, allowing for cross-case comparisons, and comparisons to previous works.


Extra loss due to Fano resonances in inhibited coupling fibers based on a lattice of tubes.
Confinement loss of inhibited coupling fibers with a cladding composed of a lattice of tubes of various shapes is theoretically and numerically investigated. Both solid core and hollow core are taken into account. It is shown that in case of polygonal shaped tubes, confinement loss is affected by extra loss due to Fano resonances between core modes and cladding modes with high spatial dependence. This explains why hollow core Kagome fibers exhibit much higher confinement loss with respect to tube lattice fibers and why hypocycloid core cladding interfaces significantly reduce fiber loss. Moreover it is shown that tube deformations, due for example to fabrication process, affect fiber performances. A relationship between the number of polygon sides and the spectral position of the extra loss is found. This suggests general guide lines for the design and fabrication of fibers free of Fano resonance in the spectral range of interest.


Two types of slip-induced falls among community dwelling older adults.
Little is known about the landing behavior of the trailing (recovery) foot and ensuing types of falls following a forward slip in walking. The purposes of this study were to (1) determine if community-dwelling older adults experienced bilateral slips at the same rate as had been previously observed for young adults during over-ground walking; (2) determine if fall rate in older adults was dependent on slip type (unilateral vs. bilateral); and (3) identify differences in spatiotemporal variables of the trailing leg step between unilateral and bilateral slips. One-hundred-seventy-four participants experienced an unannounced, unrehearsed slip while walking on a 7-m walkway. Each trial was monitored with a motion capture system and bilateral ground reaction force plates. Although the experimental design, developed with original data from a young adult population, favored bilateral slips, more older adults (35%) than anticipated (10% previously observed in young, p<0.001) displayed a unilateral slip. The probability of fall was equal in the two types of slips. Eighty-two people recovered from the slip, while the remaining 92 (53%) fell. These 92 were classified into two exclusive categories based on the heel distance at the time of fall arrest using cluster analysis: those which resembled a fall into a "splits" position (n=47) or a feet-forward fall (n=45). All (100%) unilateral slips led to splits falls, as expected. Yet, not all bilateral slips (only 83%) resulted in feet-forward falls. A longer forward recovery step with a prolonged step time led to both feet slipping, nearly together, hence a feet-forward fall.


The RXL motif of the African cassava mosaic virus Rep protein is necessary for rereplication of yeast DNA and viral infection in plants.
Geminiviruses, single-stranded DNA plant viruses, encode a replication-initiator protein (Rep) that is indispensable for virus replication. A potential cyclin interaction motif (RXL) in the sequence of African cassava mosaic virus Rep may be an alternative link to cell cycle controls to the known interaction with plant homologs of retinoblastoma protein (pRBR). Mutation of this motif abrogated rereplication in fission yeast induced by expression of wildtype Rep suggesting that Rep interacts via its RXL motif with one or several yeast proteins. The RXL motif is essential for viral infection of Nicotiana benthamiana plants, since mutation of this motif in infectious clones prevented any symptomatic infection. The cell-cycle link (Clink) protein of a nanovirus (faba bean necrotic yellows virus) was investigated that activates the cell cycle by binding via its LXCXE motif to pRBR. Expression of wildtype Clink and a Clink mutant deficient in pRBR-binding did not trigger rereplication in fission yeast.


Anti-CD54 (ICAM-1) has antitumor activity in SCID mice with human myeloma cells.
Recent studies have suggested that ICAM-1 (CD54) is involved in the pathogenesis of human multiple myeloma. A monoclonal antihuman CD54 antibody has been generated by immunizing BALB/c mice with human myeloma cell lines. SCID mice injected with human ARH-77 myeloma cells develop disseminated myeloma which is similar in several respects to multiple myeloma in humans. The mice have monoclonal gammopathy and succumb to hind leg paralysis caused by infiltration of tumor cells into the thoracolumbar vertebrae, resulting in compression of the spinal cord. In the absence of treatment, the mean paralysis time of the SCID/ARH-77 mice is 29 days. When the SCID/ARH-77 mice received four consecutive daily i.v. injections of anti-CD54 mAb commencing 1 day after tumor inoculation, they survived for 150 days, at which time the experiment was terminated. Histopathological analyses indicated that prior to death all control SCID/ARH-77 mice had myeloma cells in the vertebrae and skull. At this time, the anti-CD54-treated mice had no evidence of tumor. High levels of human immunoglobulin were detected in the sera of control, but not treated mice. F(ab')2 fragments of the anti-CD54 antibody also had similar, albeit, slightly less antitumor activity in vivo, suggesting that antibody effector function may account for some, but not all the antitumor activity of anti-CD54. In vitro studies indicate that anti-CD54 does not inhibit homotypic adhesion, the binding of myeloma cells to murine bone marrow stromal cells, or cell proliferation. By exclusion, we propose that the CD54-mediated homing of these ARH-77 cells to certain anatomical sites is crucial for their growth in vivo.


Comparison of utilization of dental care services among Chinese- and Russian-speaking immigrant elders.
The purpose of the study was to identify factors predictive of use of dental services among Chinese- and Russian-speaking immigrant elders. The data for this analysis were collected from the 1997 survey "Assessing public health and health care needs of Russian-speaking elderly immigrants." A similar survey was replicated among Chinese-speaking elderly immigrants in 2000. Community-based samples of 300 Russian elders and 177 Chinese elders were recruited for the study. Chinese elders used dental care services at lower rates than Russian elders. Education, length of stay in the U.S., social support, and smoking behavior were significant predictors for the use of dental services among Chinese. However, among Russian elders, age, income, and denture use predicted utilization of dental services. Although Chinese- and Russian-speaking elders have similar immigrant experiences and share the same geographic location and urban setting, the two groups have different patterns of dental service use. These differences may be due to differences in socio-demographic characteristics, values, attitudes and knowledge of oral health and dental care, and unique cultural backgrounds.


Signalling via the osteopontin and high mobility group box-1 axis drives the fibrogenic response to liver injury.
Liver fibrosis is associated with significant collagen-I deposition largely produced by activated hepatic stellate cells (HSCs); yet, the link between hepatocyte damage and the HSC profibrogenic response remains unclear. Here we show significant induction of osteopontin (OPN) and high-mobility group box-1 (HMGB1) in liver fibrosis. Since OPN was identified as upstream of HMGB1, we hypothesised that OPN could participate in the pathogenesis of liver fibrosis by increasing HMGB1 to upregulate collagen-I expression. Patients with long-term hepatitis C virus (HCV) progressing in disease stage displayed enhanced hepatic OPN and HMGB1 immunostaining, which correlated with fibrosis stage, whereas it remained similar in non-progressors. Hepatocyte cytoplasmic OPN and HMGB1 expression was significant while loss of nuclear HMGB1 occurred in patients with HCV-induced fibrosis compared with healthy explants. Well-established liver fibrosis along with marked induction of HMGB1 occurred in CCl4-injected OpnHep transgenic yet it was less in wild type and almost absent in Opn-/- mice. Hmgb1 ablation in hepatocytes (Hmgb1ΔHep) protected mice from CCl4-induced liver fibrosis. Coculture with hepatocytes that secrete OPN plus HMGB1 and challenge with recombinant OPN (rOPN) or HMGB1 (rHMGB1) enhanced collagen-I expression in HSCs, which was blunted by neutralising antibodies (Abs) and by Opn or Hmgb1 ablation. rOPN induced acetylation of HMGB1 in HSCs due to increased NADPH oxidase activity and the associated decrease in histone deacetylases 1/2 leading to upregulation of collagen-I. Last, rHMGB1 signalled via receptor for advanced glycation end-products and activated the PI3K-pAkt1/2/3 pathway to upregulate collagen-I. During liver fibrosis, the increase in OPN induces HMGB1, which acts as a downstream alarmin driving collagen-I synthesis in HSCs.


Exposure to methylene chloride. II. Psychological functions.
The effect of the solvent methylene chloride on psychological functions such as reaction time, short-term memory, and numerical ability was studied in 14 healthy male subjects. Each subject was tested repeatedly during exposure to 870, 1,740, 2,600, and 3,470 mg/m3 of methylene choride in inspiratory air and during control conditions. Samples of sub ject alveolar air were taken every third minute during the experiments. No statistically significant impairement in the performance of examined functions could be observed during exposure to methylene choride as compared to control conditions. Inexposure to the highest concentration a greater irregularity of response was obtained for subject reaction time than under control conditions.


The orphan drug dichloroacetate reduces amyloid beta-peptide production whilst promoting non-amyloidogenic proteolysis of the amyloid precursor protein.
The amyloid cascade hypothesis proposes that excessive accumulation of amyloid beta-peptides is the initiating event in Alzheimer's disease. These neurotoxic peptides are generated from the amyloid precursor protein via sequential cleavage by β- and γ-secretases in the 'amyloidogenic' proteolytic pathway. Alternatively, the amyloid precursor protein can be processed via the 'non-amyloidogenic' pathway which, through the action of the α-secretase a disintegrin and metalloproteinase (ADAM) 10, both precludes amyloid beta-peptide formation and has the additional benefit of generating a neuroprotective soluble amyloid precursor protein fragment, sAPPα. In the current study, we investigated whether the orphan drug, dichloroacetate, could alter amyloid precursor protein proteolysis. In SH-SY5Y neuroblastoma cells, dichloroacetate enhanced sAPPα generation whilst inhibiting β-secretase processing of endogenous amyloid precursor protein and the subsequent generation of amyloid beta-peptides. Over-expression of the amyloid precursor protein partly ablated the effect of dichloroacetate on amyloidogenic and non-amyloidogenic processing whilst over-expression of the β-secretase only ablated the effect on amyloidogenic processing. Similar enhancement of ADAM-mediated amyloid precursor protein processing by dichloroacetate was observed in unrelated cell lines and the effect was not exclusive to the amyloid precursor protein as an ADAM substrate, as indicated by dichloroacetate-enhanced proteolysis of the Notch ligand, Jagged1. Despite altering proteolysis of the amyloid precursor protein, dichloroacetate did not significantly affect the expression/activity of α-, β- or γ-secretases. In conclusion, dichloroacetate can inhibit amyloidogenic and promote non-amyloidogenic proteolysis of the amyloid precursor protein. Given the small size and blood-brain-barrier permeability of the drug, further research into its mechanism of action with respect to APP proteolysis may lead to the development of therapies for slowing the progression of Alzheimer's disease.


Differential Shannon entropy and differential coefficient of variation: alternatives and augmentations to differential expression in the search for disease-related genes.
Differential expression has been a standard tool for analysing case-control transcriptomic data since the advent of microarray technology. It has proved invaluable in characterising the molecular mechanisms of disease. Nevertheless, the expression profile of a gene across samples can be perturbed in ways that leave the expression level unaltered, while a biological effect is nonetheless present. This paper describes and analyses differential Shannon entropy and differential coefficient of variation, two alternate techniques for identifying genes of interest. Ontological analysis across 16 human disease datasets demonstrates that these alternatives are effective at identifying disease-related genes not found by mere differential expression alone. Because the two alternate techniques are based on somewhat different mathematical formulations, they tend to produce somewhat different gene lists. Moreover, each may pinpoint genes completely overlooked by the other. Thus, measures of entropy and variation can be used to replace or better yet augment standard differential expression computations.


Outbreak of pseudoinfection with Tsukamurella paurometabolum traced to laboratory contamination: efficacy of joint epidemiological and laboratory investigation.
From January 1988 to May 1989, one hospital in South Carolina reported 12 isolates of Tsukamurella paurometabolum from 10 patients. There were no common risk factors among the patients. Case-control studies revealed that the positive specimens were significantly more likely to have been processed in the TB/fungal room, to have been tissue samples, and to have been handled by one technician. Typing on the basis of biochemical, antimicrobial resistance, Southern blot, and ribotype profiles showed that the isolates from the outbreak were essentially identical and that they were distinguishable from each of two isolates obtained after the outbreak and from two type strains. These findings support the hypothesis of a common-source outbreak of pseudoinfection. There are reasons to believe that T. paurometabolum is present both in the environment and as a culture contaminant more often than has been recognized and that it is very rarely the true cause of infection in humans. Typing results show differences between one type strain and all of the other isolates studied in terms of colonial morphology, biochemistry, antimicrobial susceptibility, and ribotyping; these differences suggest that the nomenclature of T. paurometabolum may require further clarification.


Effects of electrical stimulation of vastus medialis obliquus muscle in patients with patellofemoral pain syndrome: an electromyographic analysis.
The use of surface electromyography (SEMG) has been considered a tool for quantitative assessment of patellofemoral pain syndrome (PFPS). Conservative treatments aim to improve patellar alignment, and electrical stimulation of the vastus medialis obliquus (VMO) muscle has been considered effective because it is selective and does not cause joint irritation. This study aims to investigate the efficiency of a muscle strengthening program with electrical stimulation of the VMO muscle in PFPS by SEMG. A group of ten young women (age: 23.1 ± 4.9 years; body mass: 66.8 ± 14.0 kg; height: 1.63 ± 6.9 cm; BMI: 25.1 ± 5.6 kg/m²) with unilateral PFPS participated in the study. They performed the functional test of stair stepping to capture the electromyographic (EMG) activity of the VMO and vastus lateralis (VL) muscles, before and after a program of electrical stimulation of the VMO muscle. The electrical stimulation was performed three times per week for six weeks. For analysis between the VMO and VL muscles, we considered the variables: ratio of time of onset to peak of activation, ratio of the integrals of the signals (t-test for dependent samples), and difference between onsets of activation (Wilcoxon test), with significance level of p < 0.05. The results only showed change in behavior in the EMG signal for the ratio of the integrals of the signals, indicating that changes occurred in the force-generating capacity of the muscle after the training. The use of electrical stimulation should be considered to complement the conservative therapeutic approach in patients with PFPS, and the analysis of the ratio of the integrals of the SEMG signals should be considered as an instrument of evaluation. Article registered in the Australian New Zealand Clinical Trials Registry (ANZCTR) under number ACTRN 12609000079246.


Biofuels: network analysis of the literature reveals key environmental and economic unknowns.
Despite rapid growth in biofuel production worldwide, it is uncertain whether decision-makers possess sufficient information to fully evaluate the impacts of the industry and avoid unintended consequences. Doing so requires rigorous peer-reviewed data and analyses across the entire range of direct and indirect effects. To assess the coverage of scientific research, we analyzed over 1600 peer-reviewed articles published between 2000 and 2009 that addressed 23 biofuels-related topics within four thematic areas: environment and human well-being, economics, technology, and geography. Greenhouse gases, fuel production, and feedstock production were well-represented in the literature, while trade, biodiversity, and human health were not. Gaps were especially striking across topics in the Southern Hemisphere, where the greatest potential socio-economic benefits, as well as environmental damages, may co-occur. There was strong asymmetry in the connectedness of research topics; greenhouse gases articles were twice as often connected to other topics as biodiversity articles. This could undermine the ability of scientific and economic analyses to adequately evaluate impacts and avoid significant unintended consequences. At the least, our review suggests caution in this developing industry and the need to pursue more interdisciplinary research to assess complex trade-offs and feedbacks inherent to an industry with wide-reaching potential impacts.


microRNA-145 Mediates the Inhibitory Effect of Adipose Tissue-Derived Stromal Cells on Prostate Cancer.
Adipose-derived stromal cell (ASC), known as one of the mesenchymal stem cells (MSCs), is a promising tool for regenerative medicine; however, the effect of ASCs on tumor growth has not been studied sufficiently. We investigated the hypothesis that ASCs have an inhibitory effect on metastatic tumor progression. To evaluate the in vitro inhibitory effect of ASCs on metastatic prostate cancer (PCa), direct coculture and indirect separate culture experiments with PC3M-luc2 cells and human ASCs were performed, and ASCs were administered to PC3M-luc2 cell-derived tumor-bearing nude mice for in vivo experiment. We also performed exosome microRNA (miRNA) array analysis to explore a mechanistic insight into the effect of ASCs on PCa cell proliferation/apoptosis. Both in vitro and in vivo experiments exhibited the inhibitory effect of ASCs on PC3M-luc2 cell proliferation, inducing apoptosis and PCa growth, respectively. Among upregulated miRNAs in ASCs compared with fibroblasts, we focused on miR-145, which was known as a tumor suppressor. ASC-derived conditioned medium (CM) significantly inhibited PC3M-luc2 cell proliferation, inducing apoptosis, but the effect was canceled by miR-145 knockdown in ASCs. ASC miR-145 knockdown CM also reduced the expression of Caspase 3/7 with increased antiapoptotic protein, BclxL, expression in PC3M-luc2 cells. This study provides preclinical data that ASCs inhibit PCa growth, inducing PCa cell apoptosis with reduced activity of BclxL, at least in part, by miR-145, including exosomes released from ASCs, suggesting that ASC administration could be a novel and promising therapeutic strategy in patients with PCa.


Live cell tagging tracking and isolation for spatial transcriptomics using photoactivatable cell dyes.
A cell's phenotype and function are influenced by dynamic interactions with its microenvironment. To examine cellular spatiotemporal activity, we developed SPACECAT-Spatially PhotoActivatable Color Encoded Cell Address Tags-to annotate, track, and isolate cells while preserving viability. In SPACECAT, samples are stained with photocaged fluorescent molecules, and cells are labeled by uncaging those molecules with user-patterned near-UV light. SPACECAT offers single-cell precision and temporal stability across diverse cell and tissue types. Illustratively, we target crypt-like regions in patient-derived intestinal organoids to enrich for stem-like and actively mitotic cells, matching literature expectations. Moreover, we apply SPACECAT to ex vivo tissue sections from four healthy organs and an autochthonous lung tumor model. Lastly, we provide a computational framework to identify spatially-biased transcriptome patterns and enriched phenotypes. This minimally perturbative and broadly applicable method links cellular spatiotemporal and/or behavioral phenotypes with diverse downstream assays, enabling insights into the connections between tissue microenvironments and (dys)function.


Isokinetic and isometric muscle strength combined with transcutaneous electrical muscle stimulation in primary fibromyalgia syndrome.
Twenty women with primary fibromyalgia syndrome and 20 age matched healthy women were investigated. The subjects performed maximum voluntary isokinetic contractions of the right quadriceps in an isokinetic dynamometer. Maximum voluntary isometric contractions of the right quadriceps were performed with superimposed transcutaneous electrical stimulation. The examination protocol was repeated after 1 h of resting. Isokinetic and isometric muscle strength was found to be, respectively, 45% (p = 0.0001) and 44% (p = 0.0001) lower in the patient group compared to the healthy subjects. The frequency of superimposed twitches was 65% in the patient group and 15% in the control group (p = 0.003). Patients with primary fibromyalgia have a lower maximum voluntary muscle strength than expected. The increased presence of superimposed electrically elicited twitches during maximum voluntary contraction indicates submaximal force application in primary fibromyalgia syndrome.


Effects of epidermal growth factor on fibroblast migration through biomimetic hydrogels.
We have previously reported on the development and use of synthetic hydrogel extracellular matrix (ECM) analogues that can be used to study the mechanisms of migration. These biomimetic hydrogels consist of bioinert poly(ethylene glycol) diacrylate derivatives with proteolytically degradable peptide sequences included in the backbone of the polymer and adhesion peptide sequences grafted into the network. Cells adhere to the hydrogel via interaction between the grafted adhesion ligands and receptors on the cell surface. The cells migrate through the three-dimensional system by secreting the appropriate proteolytic enzymes, which are involved in cell migration and are targeted to the peptide sequences incorporated in the backbone of the polymer. It was observed that cell migration has a biphasic dependence on adhesion ligand concentration, with optimal migration at intermediate ligand levels. In this study, we demonstrate that we can covalently attach epidermal growth factor (EGF) to PEG and graft them into the hydrogels. It was observed that EGF when tethered maintained mitogenic activity. It was also observed that fibroblast migration significantly increased in the presence of the grafted EGF through the collagenase-sensitive hydrogels. In addition, the increase in migration was found to be independent from the proliferative response of the cells. These synthetic ECM analogues allow one to systematically control identities and concentrations of biomolecules and are useful tools to study mechanisms of cell migration.


The origin of modern terrestrial life.
The study of the origin of life covers many areas of expertise and requires the input of various scientific communities. In recent years, this research field has often been viewed as part of a broader agenda under the name of "exobiology" or "astrobiology." In this review, we have somewhat narrowed this agenda, focusing on the origin of modern terrestrial life. The adjective "modern" here means that we did not speculate on different forms of life that could have possibly appeared on our planet, but instead focus on the existing forms (cells and viruses). We try to briefly present the state of the art about alternative hypotheses discussing not only the origin of life per se, but also how life evolved to produce the modern biosphere through a succession of steps that we would like to characterize as much as possible.


Construct and concurrent validation of the MG-QOL15 in the practice setting.
Health-related quality of life (HRQOL) estimates can play an important role in patient care by providing information about the patient's perception of impairment and disability and the degree to which the patient tolerates disease manifestations. The 15-item myasthenia gravis quality of life scale (MG-QOL15) is an HRQOL evaluative instrument specific to patients with myasthenia gravis (MG) that was designed to be easy to administer and interpret. In this multicenter study we demonstrate the construct validity of the MG-QOL15 in the practice setting. To assess the construct validity, score distributions were examined for test items in different MG patient groups that represent the clinical spectrum of the disease. For example, patients in remission more frequently scored test items as normal than did patients in other groups. Patients with lower (better) MG composite scores also more frequently scored items as normal than did patients with higher (worse) scores. There was also appropriate correlation between the MG-QOL15 and the other MG-specific scales studied. The study findings shed light on what troubles MG patients. The MG-QOL15 has construct validity in the clinical practice setting and represents an efficient and valuable tool for assessing HRQOL for patients with MG.


Hyperglycemia after intense exercise in IDDM subjects during continuous subcutaneous insulin infusion.
Exercise is conventionally considered a modality for improvement of glycemia in diabetes. We have found that a short period of intense exercise (80% VO2max) in normal lean subjects produces sustained postexercise hyperglycemia 20% above basal with a corresponding 100% increase in plasma insulin. In people with insulin-dependent diabetes mellitus (IDDM) incapable of this insulin response, it was predicted that postexercise hyperglycemia would be of greater magnitude and/or duration. To investigate this possibility, the effects of the same intense exercise (80% VO2max) were studied in 8 IDDM subjects (2 on 2 occasions) in the postabsorptive state with continuous subcutaneous (abdominal) insulin infusion (CSII). When the preexercise plasma glucose was normal (n = 6, 86 +/- 4 mg/dl), there ensued a postexercise hyperglycemia to 127 +/- 7 mg/dl (P less than .001) sustained for 2 h postexhaustion. Plasma free immunoreactive insulin (IRI) was 1.43 +/- 0.12 ng/ml before exercise and did not change postexercise. When mean preexercise plasma glucose was 149 +/- 9 mg/dl (n = 4), it rose progressively throughout the 2 h of recovery to 229 +/- 28 mg/dl (P less than .025). A small but statistically significant decrease in free IRI occurred during the last 80 min of recovery. Hyperglycemia in the diabetic subjects was not explained by abnormal or differing responses of glucagon or catecholamines. Thus, with intense exercise, diabetic control deteriorates rather than improves. Therefore, different therapeutic strategies may be required for intense compared with moderate exercise in IDDM patients.


All roads lead to the motor cortex: psychomotor mechanisms and their biochemical modulation in psychiatric disorders.
Psychomotor abnormalities have been abundantly observed in psychiatric disorders like major depressive disorder (MDD), bipolar disorder (BD), and schizophrenia (SCH). Although early psychopathological descriptions highlighted the truly psychomotor nature of these abnormalities, more recent investigations conceive them rather in purely motor terms. This has led to an emphasis of dopamine-based abnormalities in subcortical-cortical circuits including substantia nigra, basal ganglia, thalamus, and motor cortex. Following recent findings in MDD, BD, and SCH, we suggest a concept of psychomotor symptoms in the literal sense of the term by highlighting three specifically psychomotor (rather than motor) mechanisms including their biochemical modulation. These include: (i) modulation of dopamine- and substantia nigra-based subcortical-cortical motor circuit by primarily non-motor subcortical raphe nucleus and serotonin via basal ganglia and thalamus (as well as by other neurotransmitters like glutamate and GABA); (ii) modulation of motor cortex and motor network by non-motor cortical networks like default-mode network and sensory networks; (iii) global activity in cortex may also shape regional distribution of neural activity in motor cortex. We demonstrate that these three psychomotor mechanisms and their underlying biochemical modulation are operative in both healthy subjects as well as in MDD, BD, and SCH subjects; the only difference consists in the fact that these mechanisms are abnormally balanced and thus manifest in extreme values in psychiatric disorders. We conclude that psychomotor mechanisms operate in a dimensional and cross-nosological way as their degrees of expression are related to levels of psychomotor activity (across different disorders) rather than to the diagnostic categories themselves. Psychomotor mechanisms and their biochemical modulation can be considered paradigmatic examples of a dimensional approach as suggested in RDoC and the recently introduced spatiotemporal psychopathology.


Three-dimensional photographic analysis of the face in European adults from southern Spain with normal occlusion: reference anthropometric measurements.
Recent non-invasive 3D photography method has been applied to facial analysis, offering numerous advantages in orthodontic. The purpose of this study was to analyze the faces of a sample of healthy European adults from southern Spain with normal occlusion in order to establish reference facial soft tissue anthropometric parameters in this specific geographic-ethnic population, as well as to analyze sexual dimorphism. A sample of 100 healthy adult volunteers consisting of 50 women (mean age, 22.92 ± 1.56 years) and 50 men (mean age, 22.37 ± 2.12 years) were enrolled in this study. All participants had normal occlusion, skeletal Class I, mesofacial pattern, and healthy body mass index. Three-dimensional photographs of the faces were captured non-invasively using Planmeca ProMax 3D ProFace®. Thirty landmarks related to the face, eyes, nose, and orolabial and chin areas were identified. Male displayed higher values in all vertical and transversal dimensions, with the exception of the lower lip height. Larger differences between sexes were observed in face, mandible, and nose. Male also had higher values in the angular measurements which referred to the nose. No sex differences were found in transverse upper lip prominence or transverse mandibular prominence. No differences were found in the ratio measurements, with the exception of intercantal width/nasal width, which was higher in women than in men. Reference anthropometric measurements of facial soft tissues have been established in European adults from southern Spain with normal occlusion. Significant sexual dimorphism was found, with remarkable differences in size between sexes.


CHANGES OF THE FOVEAL AVASCULAR ZONE AND MACULAR MICROVASCULATURE WITHIN THE FRAMEWORK OF OCT ANGIOGRAPHY EXAMINATION IN YOUNG PATIENTS WITH TYPE 1 DIABETES (PILOT STUDY).
Learn about the development and changes in foveal avascular zone (FAZ) and vascularity of retina in the surrounding zone, depending on the duration in young diabetic patients type 1 (T1DM). As part of regular one-year examinations of young T1DM patients at the Eye Clinic of the University Hospital Královské Vinohrady in Prague (Czech Republic, EU) from January to December 2019, OCT angiography using the device Spectralis (Heidelberg Engineering) was included. Forty patients aged 18 to 30 years were examined, median 21 years. T1DM was diagnosed in childhood and lasted for more than 10 years. At the same time, a control group of forty individuals of similar age, without metabolic and other general disease was examined, normal visual acuity and physiological fundoscopic finding were obligatory. The FAZ size was evaluated in both groups (using built-in function "Draw Region"), also its shape, density decrease and change in character of vascularity of the retina was assessed. In the control group, the FAZ area ranged from 0.06 to 0.4 mm², with an average of 0.253 ± 0.092 mm² and a median of 0.27 mm². It was not affected by a fundamental change in its round shape and the surrounding capillary netting was regular and reasonably dense. In T1DM patients, the FAZ area was in a wider range, from 0.05 to 0.64 mm², an average of 0.300 ± 0.132 mm², and a median of 0.31 mm². The difference in FAZ across-the-board evaluation was statistically significant (p = 0, 009). Diabetic preretinopathy (DpR) was defined by the irregularity of the capillary density up to the manifestation of non-perfusion, in 61% of cases the size of the FAZ was changed. In diabetic retinopathy (DR) there was always an irregularity of the FAZ shape with its enlargement, manifestation of non-perfusion, capillary dilatation and rare microaneurysms. Changes in FAZ size corresponded to the stage of T1DM on the fundoscopic finding of the eye depending on its duration. The initial increased amount of foveal capillaries, which resulted in decreased FAZ area, was followed by a gradual decrease in capillaries and increased FAZ area, consistent with the manifestations of DpR. It was accompanied by a change in capillary density in macula to eventual non-perfusion. On the contrary, the increase in the FAZ area and its irregularity accompanied by non-perfusion of the capillary net and microaneurysms corresponded to the development of DR already.


Shigella flexneri invasion of HeLa cells induces NF-kappa B DNA-binding activity.
Although information about the genetic basis and mechanisms of Shigella flexneri cellular invasion is accumulating, little is known about changes in cell signaling and their consequences following bacterium-host cell interactions. A general result of signal transduction is alterations in the levels and/or activities of transcription factors. Alterations in transcription factor binding activities were observed following challenge with S. flexneri. Changes in the DNA-binding activities of cellular transcription factors to AP1, AP2, cyclic AMP response element, CTF1/NF1, NF-kappa B/Rel, OCT1, and SP1 DNA-binding sites were investigated by electrophoretic mobility shift assays. NF-kappa B/Rel DNA-binding activity was enhanced more than 11-fold by cellular invasion; noninvasive S. flexneri strains induced low levels of kappa B DNA binding. Both subunits of the NF-kappa B transcription factor, p50 and p65, but not c-Rel (p85), are components of the kappa B DNA-binding activity. These data suggest that changes in cellular transcription factor binding activity are a consequence of S. flexneri invasion, and these changes could play a role in the initial host response or in the pathogenesis of the disease.


Effects of loop diuretics on angiotensin II-stimulated vascular smooth muscle cell growth.
Torasemide and furosemide are diuretics that inhibit the Na(+), K(+), 2Cl(-) co-transporter localized in cells from the ascending limb of the loop of Henle. The effects of torasemide and furosemide on cell growth induced by angiotensin II (Ang II) were investigated in cultured vascular smooth muscle cells (VSMCs) obtained from the aorta of adult spontaneously hypertensive rats (SHR). Cell growth was determined by DNA and protein synthesis as measured by [3H]thymidine and [3H]leucine incorporation, respectively. Proliferation of VSMCs was measured using a non-radioactive colorimetric cell proliferation assay. Ang II (10(-7) M) signficantly increased DNA and protein synthesis and cell proliferation in VSMCS: These effects were completely abolished by the Ang II type 1 receptor antagonist irbesartan (10(-6) M). Ang II-induced [3H]leucine incorporation was reduced in a dose-dependent way by torasemide (IC(50) value: 7.7+/-0.8x10(-7) M) but not by furosemide. Neither torasemide nor furosemide modified Ang II-stimulated [3H]thymidine incorporation or proliferation in VSMCs. These results indicate that torasemide, but not furosemide, inhibits Ang II-induced protein synthesis in VSMCs from SHR. Thus, it is suggested that the capacity of torasemide to block this trophic action of Ang II in rat VSMCs is not mediated by inhibition of the Na(+), K(+), 2Cl(-) co-transport mechanism.


Reiter's keratitis.
A distinctive keratitis occurs commonly in Reiter's syndrome. In three patients with Reiter's keratitis, two demonstrated the typical features of prodromal conjunctivitis, subepithelial and anterior stromal infiltrates, ragged epithelial erosions, and spontaneous resolution. A third case of rare, severe keratitis in addition had an associated finding of disciform keratitis. To our knowledge, this last finding has not previously been reported. Chlamydia has been implicated as an etiologic agent in Reiter's syndrome. Giemsa's stain of corneal epithelial cells in one of our patients disclosed intracytoplasmic inclusions that resembled those seen in Chlamydia-caused conjunctivitis. The patient also exhibited a rising serum titer to Chlamydia antigen.


GP96 is over-expressed in oral cavity cancer and is a poor prognostic indicator for patients receiving radiotherapy.
Oral cavity cancers (ORC) are the most common cancers, and standard treatment is radical surgery with postoperative radiotherapy. However, locoregional failure remains a major problem, indicating radioresistance an important issue. Our previous work has shown that GP96 contributed to radioresistance in nasopharyngeal and oral cancer cell lines. In this study, we determined clinical significance of GP96 in ORC by evaluation of GP96 expression and its association with disease prognosis in patients receiving radiotherapy Total of 79 ORC patients (77 males, median age: 48 years old) receiving radical surgery and postoperative radiotherapy between Oct 1999 and Dec 2004 were enrolled. Patients in pathological stages II, III and IV were 16.5%, 16.5% and 67%, respectively. For each patient, a pair of carcinoma tissue and grossly adjacent normal mucosa was obtained. GP96-expression was examined by western blot analysis, and the association with clinicopathological status was determined. Three-year locoregional control (LRC), distant metastasis-free survival (DMFS), disease-specific survival (DSS) and overall survival (OS) rates were 69%, 79%, 63% and 57%, respectively. We found that 55 patients (70%) displayed GP96-overexpression in the tumor tissue, which correlated with a higher pN stage (p = 0.020) and tumor depth (> 10 mm) (p = 0.045). Nodal extracapsular spreading (ECS) and GP96-overexpression predicted adverse LRC (p = 0.049 and p = 0.008). When stratified by nodal ECS, the adverse impact of GP96 remained significant in three-year LRC (p = 0.004). In multivariate analysis, GP96-overexpression was also an independent predictor of LRC, DSS and OS (p = 0.018, p = 0.011 and p = 0.012). GP96 may play roles in radioresistance which attributes to tumor invasiveness in oral cancer patients receiving radiotherapy. GP96 may serve as a novel prognostic marker of radiotherapy. However, further independent studies are required to validate our findings in a larger series.


Azacitidine in the management of patients with myelodysplastic syndromes.
Myelodysplastic syndromes (MDS) are a heterogeneous group of clonal hematopoeitic disorders characterized by ineffective hematopoiesis and potential transformation to acute myeloid leukemia (AML). For decades, the mainstay of treatment for MDS was supportive care, including transfusion of blood products and growth factors. Further understanding of disease biology led to the discovery of a high prevalence of hypermethylation of tumor suppressor genes in high-risk MDS and secondary leukemias. Hence, the role of irreversible DNA methlytransferase inhibitors such as azacitidine was investigated with promising outcomes in the treatment of MDS. Azacitidine was initially approved in the USA by the Food and Drug Administration (FDA) in 2004 for the treatment of all subtypes of MDS and was granted expanded approval in 2009 to reflect new overall survival data demonstrated in the AZA-001 study of patients with higher-risk MDS. Azacitidine has demonstrated significant and clinically meaningful prolongation of survival in higher-risk patients with MDS and has changed the natural history of these disorders. The agent maintains a relatively safe toxicity profile, even in older patients. The role of azacitidine has been explored in the treatment of AML and chronic myelomonocytic leukemia and has also been studied in the peritransplant setting. Azacitidine has been combined with other novel agents such as lenalidomide, histone deacetylase inhibitors and growth factors in the hope of achieving improved outcomes. Currently, both intravenous and subcutaneous forms of azacitidine are approved for use in the USA with the oral form being granted fast track status by the FDA.


Inbreeding depression on production and reproduction traits of buffaloes from Brazil.
The objective of this study was to evaluate the influence of inbreeding depression on traits of buffaloes from Brazil. Specifically, the traits studied were body weight at 205 and 365 days of age, average daily gain from birth to 205 days (ADG_205), average daily gain between 205 and 365 days (ADG205_365) in Mediterranean buffaloes, and milk yield, lactation length, age of first calving and calving intervals in Murrah buffaloes. Inbreeding effects on the traits were determined by fitting four regression models (linear, quadratic, exponential and Michaelis-Menten) about the errors generated by the animal model. The linear model was only significant (P < 0.05) for growth traits (exception of ADG205_365). The exponential and Michaelis-Menten models were significant (P < 0.01) for all the studied traits while the quadratic model was not significant (P > 0.05) for any of the traits. Weight at 205 and 365 days of age decreased 0.25 kg and 0.39 kg per 1% of increase in inbreeding, respectively. The inbred animals (F = 0.25) produced less milk than non-inbred individuals: 50.4 kg of milk. Moreover, calving interval increased 0.164 days per 1% of increase in inbreeding. Interestingly, inbreeding had a positive effect on age at first calving and lactation length, decreasing age of first calving and increasing lactation length.


Antibiotic prophylaxis in cataract surgery - An evidence-based approach.
Various protocols are being followed for endophthalmitis prophylaxis in cataract surgery, and this subject continues to be a matter of debate. We summarize the most recent evidence-based studies on this topic with additional stress on intracameral (IC) antibiotic prophylaxis. Here, we discuss several large, international clinical studies which discuss the efficacy, adoption, safety, cost, and newer trends in antibiotic prophylaxis. Majority of these studies report a significant reduction in endophthalmitis rates with IC antibiotic prophylaxis. Efficacy data have been reported for IC cefuroxime, vancomycin, and moxifloxacin. Surgeons are now looking for alternatives to vancomycin for IC prophylaxis because of its association with the rare but sight-threatening complication of hemorrhagic occlusive retinal vasculitis. A recent large clinical study shows convincing efficacy with IC moxifloxacin prophylaxis. Two large studies have also reported significant endophthalmitis reduction following use of IC antibiotic prophylaxis, in eyes with posterior capsule tear which are at highest risk for infection. Except for one randomized controlled trial, there is a lack of prospective data on this subject; however, considering the complexity of performing such studies, surgeons have to rely on the mounting evidence from other recent big data studies. Availability of approved intraocular antibiotic formulations will see a much higher adoption in the future.


Parthenolide-induced apoptosis, autophagy and suppression of proliferation in HepG2 cells.
To investigate the anticancer effects and underlying mechanisms of parthenolide on HepG2 human hepatocellular carcinoma cells. Cell viability was assessed by MTT assay and cell apoptosis through DAPI, TUNEL staining and Western blotting. Monodansylcadaverin(MDC) and AO staining were used to detect cell autophagy. Cell proliferation was assessed by Ki67 immunofluorescence staining. Parthenolide induced growth inhibition in HepG2 cells. DAPI and TUNEL staining showed that parthenolide could increase the number of apoptotic nuclei, while reducing the expression of the anti-apoptotic protein Bcl-2 and elevating the expression of related proteins, like p53, Bax, cleaved caspase9 and cleaved caspase3. Parthenolide could induce autophagy in HepG2 cells and inhibited the expression of proliferation-related gene, Ki-67. Parthenolide can exert anti-cancer effects by inducing cell apoptosis, activating autophagy and inhibiting cell proliferation.


Workplace violence by specialty among Peruvian medical residents.
To determine the prevalence of workplace violence among Peruvian medical residents and to evaluate the association between medical specialty and workplace violence per type of aggressor. This was a cross-sectional secondary analysis that used data from the Peruvian Medical Residents National Survey 2016 (ENMERE-2016). The outcome of interest was workplace violence, including physical and verbal violence, which were categorized according to the perpetrator of violence (patients/relatives and worker-to-worker). Primary exposure was the medical specialty, categorized as clinical, surgical, and other specialties. To evaluate the associations of interest, we estimated adjusted prevalence ratios (PR) with their respective 95% confidence intervals (95% CI) using Poisson regression models with robust variances. A total of 1054 Peruvian medical residents were evaluated. The mean age was 32.6 years and 42.3% were female. Overall 73.4% reported having suffered of workplace violence sometime during the residency, 34.4% reported violence from patients/relatives, and 61.1% reported worker-to-worker violence. Compared with clinical residents, surgical residents had a lower prevalence of violence from patients/relatives (PR: 0.71; 95% CI: 0.59-0.87), but a higher prevalence of worker-to-worker violence (PR: 1.11, 95% CI: 1.01-1.23). Nearly three quarters of medical residents reported having suffered workplace violence sometime during their residency. Compared with clinical residents, surgical residents had lower rates of violence from patients/relatives, but higher rates of worker-to-worker violence; while residents from non-clinical and non-surgical specialties had a lower prevalence of both types of violence.


Diagnosis of pulmonary problems in HIV-infected patients.
During the past 13 yrs it has become evident that pulmonary complications occur frequently and are often life-threatening in patients infected with the human immunodeficiency virus (HIV). Moreover, the spectrum of HIV-related pulmonary diseases has enlarged considerably and it now includes numerous infectious and non-infectious conditions. Because clinical and radiological presentation is generally nonspecific and often altered by diminished signs of inflammation or prophylactic measures, and multiple simultaneous complications frequently occur, an early aetiological diagnosis is essential. The diagnostic work-up of these patients must be straightforward. Examination of (induced) sputum and bronchoalveolar lavage fluid are the most important procedures. If they are not diagnostic, transbronchial biopsy, transbronchial needle aspiration, computed tomography and echocardiography are most often revealing. However, we must be continuously prepared to face new complications in the course of this devastating disease.


Quantum dot photosensitizers. Interactions with transition metal centers.
Semiconductor quantum dots (QDs) are attractive for potential use as photosensitizers for a variety of applications. These nanomaterials have very high absorption cross-sections and often display strong photoluminescence (PL). Furthermore, QD absorption and emission spectra can be tuned simply by varying their size, and QD surfaces can be modified to access multiple sites for attaching potential acceptors as well as other functionalities. Here we provide an overview of recent studies concerned with the photosensitization of transition metal centers and other acceptors. Particular focus is directed towards potential therapeutic applications and to our own interest in the delivery of small molecule bioregulators to physiological targets. Studies that have addressed factors that control likely energy and charge transfer processes between QD donors and acceptor molecules are also discussed. Understanding the mechanisms of these photosensitization processes can provide design guidelines for successful applications.