Acceleration of oxime-induced reactivation of organophosphate-inhibited fetal bovine serum acetylcholinesterase by monoquaternary and bisquaternary ligands.
Reactivation of organophosphate (OP)-inhibited acetylcholinesterase (AChE) by oximes is the primary reason for their effectiveness in the treatment of OP poisoning. Reactivation is reported to accelerate by quaternary ligands such as decamethonium, which is devoid of nucleophilicity. The mechanism of this enhancement is not known. To better understand the acceleration phenomenon, we examined ligand modulations of oxime-induced reactivation of methylphosphonylated AChE using 7-(methylethoxyphosphinyloxy)-1-methylquinolinium iodide and fetal bovine serum AChE. Edrophonium, decamethonium, and propidium, three quaternary AChE ligands of different types, were tested as potential accelerators. Experiments were carried out with both soluble enzyme preparation and AChE conjugated to polyurethane. Kinetic measurements with oximes 2-[hydroxyiminomethyl]-1-methylpyridinium chloride, 1,1'-trimethylene bis-(4-hydroxyimino methyl)-pyridinium dibromide, and 1, 1'-[oxybis-methylene)bis[4-(hydroxyimino)methyl]pyridiniu um dichloride showed that in the presence of 50 microM edrophonium, the reactivation rate constants increased 3.3-12.0-fold; 200 microM decamethonium produced a 1.6-3.0-fold enhancement of reactivation rate constants by the same oximes. Reactivation of the inhibited enzyme by 1-(2-hydroxyiminomethyl-1-pyridinium)-1-(4-carboxy-aminopyridinium )-d imethyl ether hydrochloride, 1-(2-hydroxyiminomethyl-1-pyridinium)-1-(3-carboxy-aminopyridinium )-d imethyl ether hydrochloride, and 1-[[[4-(aminocarbonyl)pyridino]methoxy]methyl]-2, 4, -bis(hydroxyimino)methyl pyridinium dichloride was not affected by either ligand. Propidium slowed the reactivation of 7-(methylethoxyphosphinyloxy)-1- methylquinolinium iodide-inhibited AChE by all oximes. Results suggest that the accelerator site may reside inside the catalytic gorge rather than at its entrance and acceleration may be due to the prevention of reinhibition of the regenerated enzyme by the putative product, the phosphonylated oxime. In addition to the nucleophilic property of the oximate anion, some of the reactivators may carry an accelerating determinant, as characterized with respect to edrophonium and decamethonium. Results offer possible explanations for the superiority of 1-(2-hydroxyiminomethyl-1-pyridinium)-1-(4-carboxy-aminopyridinium )-d imethyl ether hydrochloride over other oximes in the reactivation of specific AChE-OP conjugates.


Different effects of polymorphisms of tumor necrosis factor-alpha and interleukin-1 beta on development of peptic ulcer and gastric cancer.
In Western countries, polymorphism of pro-inflammatory cytokine genes is associated with the development of gastric cancer and duodenal ulcer. The aim of this study was to clarify the association of polymorphisms of interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha with susceptibility to peptic ulcer diseases and gastric cancer in Japan. The IL-1beta-511/-31 and TNF-alpha-308/-857/-863/-1031 genotypes were determined in Helicobacter pylori-positive patients with gastritis only (n = 164), gastric ulcers (n = 110), duodenal ulcers (n = 94), or gastric cancers (n = 105), and in H. pylori-negative controls (n = 172). Carriage of the alleles TNF-alpha-857 T (odd ratio [OR], 1.826; 95% confidence interval [CI], 1.097-3.039), TNF-alpha-863 A (OR, 1.788; 95% CI, 1.079-2.905) and TNF-alpha-1031 C (OR, 1.912; 95% CI, 1.152-3.171) was associated with increased risk for gastric ulcer development. Carriage of the alleles TNF-alpha-857 T (OR, 1.686; 95% CI, 1.003-2.832), TNF-alpha-863 A (OR, 1.863; 95% CI, 1.118-3.107) and TNF-alpha-1031 C (OR 2.074; 95% CI, 1.244-3.457) was also associated with increased risk of gastric cancer development. There was no relationship between the development of H. pylori-related diseases and polymorphisms of IL-1beta-511/-31 and TNF-alpha-308. The simultaneous carriage of three different high-producer alleles of TNF-alpha-857/-863/-1031 significantly increased the risk of gastric ulcer (OR, 6.57; 95% CI, 2.34-18.40) and gastric cancer (OR, 5.20; 95% CI, 1.83-14.78). Polymorphisms in TNF-alpha rather than IL-1beta are associated with increased risk for gastric ulcers and gastric cancer in Japan. The simultaneous carriage of more than one high-producer allele of TNF-alpha further increased the risks for gastric ulcer and cancer.


Malaria vaccines and human immune responses.
Despite reductions in malaria episodes and deaths over the past decade, there is still significant need for more effective tools to combat this serious global disease. The positive results with the Phase III trial of RTS,S directed to the circumsporozoite protein of Plasmodium falciparum have established that a vaccine against malaria can provide partial protection to children in endemic areas, but its limited efficacy and relatively short window of protection mandate that new generations of more efficacious vaccines must be sought. Evidence shows that anti-parasite immune responses can control infection against other stages as well, but translating these experimental findings into vaccines for blood stages has been disappointing and clinical efforts to test a transmission blocking vaccine are just beginning. Difficulties include the biological complexity of the organism with a large array of stage-specific genes many of which in the erythrocytic stages are antigenically diverse. In addition, it appears necessary to elicit high and long-lasting antibody titers, address the redundant pathways of merozoite invasion, and still seek surrogate markers of protective immunity. Most vaccine studies have focused on a single or a few antigens with an apparent functional role, but this is likely to be too restrictive, and broad, multi-antigen, multi-stage vaccines need further investigation. Finally, novel tools and biological insights involving parasite sexual stages and the mosquito vector will provide new avenues for reducing or blocking malaria transmission.


Changes in Serum Nampt Levels and Its Significances in Diabetic Nephropathy Patients-The Potential Role of Nampt in T2DM with Diabetic Nephropathy.
Nicotinamide phosphoribosyltransferase(Nampt) has a variety of biological effects, which can participate in the development of inflammatory diseases, metabolism diseases and chronic kidney disease, et al. However, the role of Nampt in T2DM and DN is still controversial. The study aimed to investigate changes in serum Nampt levels and its significance in diabetic nephropathy (DN). A total of 321 type 2 diabetes (T2DM) were divided into normal albuminuria group (NA, n=110), microalbuminuria group (MA, n=106) and large amount of albuminuria group (LA, n=105). 105 healthy persons were selected forming the control group (NC, n=105).; 118 T2DM without insulin therapy were screened out as the non-insulin therapy group (NIT.; n=116). Serum Nampt and interleukin (IL)-1β were measured by enzyme-linked immunosorbent assay. The relationships among the variables were analyzed by Pearson correlation analysis and Stepwise multiple regression analysis. Serum Nampt and IL-1β in the LA group were significantly higher than other groups, and they were higher in MA than in the NA group (P<0.05). Serum Nampt was positively correlated with IL-1β and urinary albumin-to-creatinine ratio (UACR) (P <0.01), and IL-1β and UACR were independent relevant factors affecting Nampt. Serum Nampt was positively correlated with insulin resistance (HOMA-IR) but negatively correlated with the quantitative insulin sensitivity check index (QUICKI) (P <0.01) in the NIT group. We indicated that serum Nampt and IL-1β were increased accompanied by the increased UACR and the association between Nampt and insulin resistance and β-cell function in T2DM with different UACRs.


Diagnosing celiac disease by video capsule endoscopy (VCE) when esophagogastroduodenoscopy (EGD) and biopsy is unable to provide a diagnosis: a case series.
Video capsule endoscopy (VCE) is mainly used to evaluate patients with celiac disease in whom their course after diagnosis has been unfavorable and the diagnosis of adenocarcinoma, lymphoma or refractory celiac disease is entertained, but it has been suggested that VCE could replace esophagogastroduodenoscopy (EGD) and biopsy under certain circumstances. We report a single center case series of 8 patients with suspected celiac disease who were diagnosed by VCE. EGD and biopsy had been performed in 4 patients resulting in a negative biopsy, declined by 2, and contraindicated in 2 due to hemophilia and von Willebrand disease. In all patients, mucosal changes of scalloping, mucosal mosaicism and reduced folds were seen in either the duodenum or jejunum on VCE. Follow-up in 7 patients demonstrated improvement in either their serological abnormalities or their presenting clinical features on a gluten-free diet. Our case series demonstrates that VCE and the visualization of the characteristic mucosal changes of villous atrophy may replace biopsy as the mode of diagnosis when EGD is either declined or contraindicated, or when duodenal biopsies are negative and there remains a high index of suspicion. Further study is needed to clarify the role and cost of diagnosing celiac disease with VCE.


Effects of iron deficiency and iron supplementation at the host-microbiota interface: Could a piglet model unravel complexities of the underlying mechanisms?
Iron deficiency is the most prevalent human micronutrient deficiency, disrupting the physiological development of millions of infants and children. Oral iron supplementation is used to address iron-deficiency anemia and reduce associated stunting but can promote infection risk since restriction of iron availability serves as an innate immune mechanism against invading pathogens. Raised iron availability is associated with an increase in enteric pathogens, especially Enterobacteriaceae species, accompanied by reductions in beneficial bacteria such as Bifidobacteria and lactobacilli and may skew the pattern of gut microbiota development. Since the gut microbiota is the primary driver of immune development, deviations from normal patterns of bacterial succession in early life can have long-term implications for immune functionality. There is a paucity of knowledge regarding how both iron deficiency and luminal iron availability affect gut microbiota development, or the subsequent impact on immunity, which are likely to be contributors to the increased risk of infection. Piglets are naturally iron deficient. This is largely due to their low iron endowments at birth (primarily due to large litter sizes), and their rapid growth combined with the low iron levels in sow milk. Thus, piglets consistently become iron deficient within days of birth which rapidly progresses to anemia in the absence of iron supplementation. Moreover, like humans, pigs are omnivorous and share many characteristics of human gut physiology, microbiota and immunity. In addition, their precocial nature permits early maternal separation, individual housing, and tight control of nutritional intake. Here, we highlight the advantages of piglets as valuable and highly relevant models for human infants in promoting understanding of how early iron status impacts physiological development. We also indicate how piglets offer potential to unravel the complexities of microbiota-immune responses during iron deficiency and in response to iron supplementation, and the link between these and increased risk of infectious disease.


"Knife-cut" intertriginous ulcers related to herpes simplex virus in three patients.
Linear intertriginous erosions and ulcerations related to herpes simplex virus (HSV) infection have been reported in patients with underlying immunosuppression. This rare presentation of HSV seems to occur predominantly in patients undergoing treatment of hematologic malignancies and rheumatologic conditions. Herein, we report three cases of linear "knife-cut" ulcerations in patients who were not undergoing active pharmacologic immunosuppressive therapy and lacked coexisting malignancy or autoimmune disease. Close examination of the skin folds for HSV infection is warranted to rule out disseminated infection as early intervention can be lifesaving.


Glycerol-preserved arterial allografts evaluated in the infrarenal rat aorta.
Vascular transplantation has become an alternative for prosthetic grafts. Suitable storage methods for vascular allografts are therefore necessary. For small caliber arterial allografts, cryopreservation and cold storage showed discouraging results. Since glycerol preservation proved effective for the storage of skin allografts, this preservation method was investigated for vascular allografts using a rat aortic transplantation model. Glycerol-preserved allografts (GA) were transplanted to the infrarenal aorta (n = 18) in Wistar rats. A control group (n = 18) underwent immediate autotransplantation (AU) of an equal length of aorta. Cumulative graft patency at 90 days' follow-up was 93% for AU and 78% for GA (ns). No aneurysm formation was detected in both groups. Intraluminal endothelial cell coverage, integrity of the media and smooth muscle cell repopulation were comparable in both groups. Intimal thickness was less in GA than in AU and inflammatory reaction in the adventitia was diminished in GA. GA were successfully grafted with acceptable patency rates compared to autografts, while intima hyperplasia and adventitial inflammatory reaction were less.


[The role of two-dimensional echocardiography in the detection of pacemaker-induced endocarditis].
The authors analysed the data of eight patients who had to undergo open heart surgery because of pacemaker endocarditis in the last four years. Repeated surgical interventions on the pacemaker system was the most predisposing factor. The most common causative organisms were Staphylococcus aureus and Staphylococcus epidermidis. Two-dimensional echocardiography (2DE) had a decision making role in the diagnosis beside the clinical picture and blood culture. 1. Each pacemaker patient with fever must be considered to have a pacemaker endocarditis; 2. All these patients must be examined by two-dimensional echocardiography.


The glutamine debate in surgery and critical care.
Glutamine (GLN) is a versatile amino acid, long believed to have important implications in ICU and surgical patients. An extensive body of data examining GLN supplementation of TPN demonstrated a consistent signal of improved outcomes. However, recently signals of risk have come from two large-scale multicenter trials evaluating GLN (and other nutrients) at high dose and as primary pharmaconutrients, not as supplementation to complete nutrition. These trials indicate a risk of increased mortality when GLN is given to patients in shock, renal failure, and early in acute phase of critical care. Recent literature continues to confirm that low and high admission GLN levels are associated with increased ICU mortality and adverse outcomes. Further, a recent meta-analysis examined trials utilizing GLN-supplemented TPN in stabile ICU patients consistent with current clinical guidelines. This analysis showed GLN supplementation of TPN led to reduced infections, LOS and hospital mortality. Three recent meta-analyses have confirmed traditional GLN-supplemented (or 'GLN-Complemented' - providing GLN for completeness of amino acid content) TPN is safe, reduces mortality and improves outcome in surgical and ICU patients. Patients in need of TPN, burns, trauma or malignancies should continue to benefit from supplemental GLN, administered either intravenously at less than 0.35 g/kg/day or enterally at less than 0.5 g/kg/day. Further, a large trial of EN GLN supplementation in burns is ongoing. Thus, when used per guideline recommendations, the GLN story is likely still relevant to ICU outcomes and research.


Fetal heart rate response to cordocentesis and pregnancy outcome: a prospective cohort.
To evaluate fetal cardiac response to cordocentesis and whether such changes may affect pregnancy outcome. 117 singleton pregnant women requiring percutaneous trans-abdominal cordocentesis were prospectively included. Fetal heart rate was continuously evaluated by ultrasound for 1 min after completion of cordocentesis and intermittently for 20 min more. Fetal and pregnancy outcomes were analyzed by grouping fetal cardiac response to cordocentesis into bradycardia, normal heart rate and tachycardia groups. Women included in the study were 30.5+/-4.0 years old and had a gestational age of 23.7+/-2.0 weeks. Fetal blood sample obtained by cordocentesis was 3.1+/-0.8 ml. Fetal heart rate before cordocentesis was 149+/-8 beats per minute (bpm), ranging from 130 to 169 bpm. Fetal heart rate post-cordocentesis was 145+/-30 bpm (from 32 to 175 bpm). The incidence of bradycardia and tachycardia was 10.3% (n=12) and 6.0% (n=7), respectively. Fetal heart rate returned to normal levels in all cases at <or=5 minutes post-cordocentesis. In the tachycardia group, 6 cases had normal pregnancy outcome and one patient was lost to follow-up. In the bradycardia group, we observed two pregnancy terminations, one patient was lost to follow-up and nine were normal outcomes. In the group with normal heart rate (n=98), we observed one intrauterine fetal death, nine adverse fetal outcomes, eight pregnancy terminations and nine patients were lost to follow-up. Transient fetal tachycardia and bradycardia were uncommon and were not associated with adverse fetal or pregnancy outcomes.


Computational Insight into Cu-Catalyzed Csp-S Coupling to Form a Macrocyclic Alkynyl Sulfide.
Copper-catalyzed Csp-S cross-coupling is known to form rare macrocyclic alkynyl sulfides. Computational studies now suggest a mechanism for the reaction pathway. Upon formation of Cu-S species, subsequent α-addition/elimination at the ethynylic carbon affords the desired macrocycle.


The mode of action of pleuromutilin derivatives. Location and properties of the pleuromutilin binding site on Escherichia coli ribosomes.
Using equilibrium dialysis techniques it could be demonstrated that (a) the pleuromutilin derivative 14-deoxy-14[(2-diethylaminoethyl)-mercaptoacetoxy] dihydromutilin HCl binds to one site per ribosome specifically, (b) the binding constant is 1.3 times 10(7) M(-1) and (c) chloramphenicol and puromycin compete with binding of the pleuromutilin derivative. Similarly the nucleotides CpA and CpCpA also displace the unsaturated derivative of the above-mentioned pleuromutilin compound. These findings suggest that the ribosomal binding site for pleuromutilin overlaps with that for chloramphenicol and analogs of the 3'-terminus of a tRNA, like puromycin or the nucleotides CpA and CpCpA.


Microcirculatory changes during chronic adaptation to hypoxia.
Microcirculatory changes in the window chamber preparation in Syrian golden hamsters, secondary to chronic hypoxia adaptation, are presented herein. Adaptation was attained by keeping animals in a 10% oxygen environment for 1 wk and 5% the following week. The following groups were studied: group 1, adapted to chronic hypoxia and kept in a 5% oxygen environment throughout the experiment; group 2, adapted to chronic hypoxia and kept in a 21% oxygen environment 24 h before and during the experiment; and group 3, control. Adaptation caused venule enlargement and hematocrit increase (68.6 +/- 2.44 in group 1, 70 +/- 2.66 in group 2, and 43.27 +/- 2.30 in group 3; P < 0.05). Whereas heart rate decreased in adapted animals, blood pressure remained constant. Group 1 presented alkalosis, hypocapnia, and hypoxemia. The adapted groups had decreased blood flow velocity in arterioles and veins. We found no difference in microvasculature oxygen tension between groups 2 and 3; however, the number of capillaries with flow was markedly reduced in group 1 but significantly increased in group 2. Our findings suggest that, as an adaptation to hypoxia, erythropoiesis may prove beneficial by increasing blood viscosity and shear stress, leading to vasodilatation, in addition to the increase in oxygen-carrying capacity. Calculations show that oxygen extraction in the tissue of the window chamber model was significantly lowered in adapted animals breathing 5% oxygen, but was unchanged from the control when breathing 21% oxygen, even though blood hemoglobin content was increased from 14.5 +/- 0.07 g/dl at control to 21.04 +/- 1.24 g/dl in the adapted animals (P < 0.05).


In Vitro Bioaccessibility and Bioavailability of Iron from Potatoes with Varying Vitamin C, Carotenoid, and Phenolic Concentrations.
The bioaccessibility and bioavailability of iron from 12 Andean potato clones were estimated using an in vitro gastrointestinal digestion procedure and the Caco-2 cell line as a model of human intestine, with ferritin formation as a marker of iron absorption. We first showed that 63.7% (for the genotype CIP_311422.016) to 79.0% (for the genotype CIP_311575.003) of the iron is released from the potato tuber matrix during in vitro gastrointestinal digestion and is therefore available at the intestinal level. On average, 32 and 24.5% of the hydrophilic bioactive components, vitamin C and chlorogenic acid, respectively, were also bioaccessible from boiled tubers. Intestinal absorption of intrinsic iron from potato tubers could not be detected using our in vitro Caco-2 cell model. When an extrinsic source of iron (20 μM FeCl3 and 1 mM ascorbic acid) was added to the digestion mixture, iron absorption varied from 1.8 to 8% for the genotypes CIP_311422.016 and CIP_311624.021, respectively, as compared to the reference control. Principal component analysis revealed negative relationships between bioavailable iron values and phenolic concentrations, whereas vitamin C concentrations were positively associated with the ferritin values. Further controlled intervention trials would be needed to conclusively assess the bioavailability of intrinsic iron from potato tubers.


High-performance Förster resonance energy transfer-based dye-sensitized photocatalytic H2 evolution with graphene quantum dots as the homogeneous energy donor.
A high-performance dye-sensitized photocatalytic H2 evolution system was developed based on Förster resonance energy transfer (FRET) by employing water-soluble and highly photoluminescent N,S codoped graphene quantum dots (NSGQDs) as the homogeneous energy donor, erythrosin B (ErB) as the sensentizing dye, and platinum nanoparticles (Pt NPs) as the catalyst. NSGQDs absorbed high-energy photons that undergo FRET to transfer the excitation energy to the sensitizing ErB for maximizing light absorption and also served as an electron transfer and loading matrix of Pt NPs for accelarating the electron transfer; as a result, the ErB-sensitized NSGQD-Pt system afforded much higher H2 evolution activity than the NSGQD-free dye-sensitized system.


Reversion of bioluminescent bacteria (Mutatox) to their luminescent state upon exposure to organic compounds, munitions, and metal salts.
Mutatox is a new genotoxicity bioassay which uses as the endpoint the bioluminescence produced on reversion of a dark strain of the marine bacterium Vibrio fischeri +/- S9. Reversion can occur by several mechanisms, including base substitution, frame-shift, SOS induction, and DNA intercalation. For screening, Mutatox provides many advantages over the Salmonella typhimurium (Ames) assay: it requires minimal sterility, employs a shorter incubation period, and does not require culture maintenance. Eighteen organic chemicals (phenol, polynuclear aromatic hydrocarbons, nitrotoluenes, others), Na3PO4, and 4 genotoxic metals (Cu2+, Ni2+, As3+, Cd2+) were tested. Most of the organic compounds positive in S. typhimurium assays were positive in Mutatox. None of the metals was genotoxic in V. fischeri, possibly due to poor uptake from the saline medium.


Inhibitory effect of an ergoline derivative, methergoline, on growth hormone and prolactin levels in acromegalic patients.
Plasma levels of growth hormone (GH) and prolactin (PRL) were measured in twelve acromegalic patients after acute administration of an ergoline derivative (methergoline) which has been proposed as a specific serotoninergic blocking agent. The administration of methergoline (4 mg p.o.) was followed by a significant decrease in plasma GH and PRL concentrations. The administration to the same subjects of CB 154 (2.5 mg p.o.), a known stimulator of dopaminergic receptors, led to results almost superimposable to those obtained with methergoline although the suppressive effect of CB 154 on GH and PRL levels was more sustained. Also on the ground of results obtained in these patients with the use of cyproheptadine, phentolamine, or pimozide, we have concluded that methergoline inhibition of GH and PRL release is, in acromegalic patients, most probably due to a dopaminergic mechanism of action.


Two new compounds from Ganoderma lucidum.
Two pairs of new enantiomers, lucidulactones A and B (1 and 2), and two known compounds were isolated from Ganoderma lucidum. Their structures were determined by means of spectroscopic methods. The chiral HPLC was used to separate the ( - )- and (+)-antipodes of the new compounds.


A study of platelet function and morphology in a new family with May-Hegglin anomaly.
A new family with May-Hegglin anomaly is presented. 9 patients were found to be affected, namely to present thrombocytopenia, giant platelets, and leukocytes inclusion bodies. A mild to moderate hemorrhagic diathesis was present in 8 patients (easy bruising, excessive bleeding after tooth extraction, menomethrorrhagia). One patient was asymptomatic. The bleeding tendency seemed to be relatively more pronounced in those patients who have larger platelets. Bleeding time was slightly prolonged in 4 of the affected patients. Platelet aggregation to Ristocetin and serotonin release was normal; on the contrary, platelet adhesiveness was slightly decreased in all patients. Plasma Btg was investigated in 7 patients, found to be normal in 5 and elevated in 2. Platelet Btg was found to be increased in all patients investigated. The ratio between Btg and platelet number was elevated in every instance. The circulating platelet mass (Btg platelet mass microgram/ml) was investigated in 7 patients, found normal in two and decreased in the remaining three. The disorder is transmitted as an autosomal dominant trait but there seems to be a variable phenotypic expression from one patient to the other.


Long-term administration of MC4 receptor antagonist HS014 causes hyperphagia and obesity in rats.
In this study we investigated the long term effects of a potent and selective melanocortin 4 (MC4) receptor antagonist (HS014) on food intake, body weight, body composition and blood glucose levels in adult rats. HS014 was injected i.c.v. either by twice-daily injections (2 x 1 nmol) for 6 days or administered by continuous infusion with osmotic minipumps (0.16 nmol/h) for 2 weeks. The results show a considerable increase in food intake and body weight after both of the treatments without any signs of tachyphylaxis. After 2 weeks of treatment with osmotic pumps, the HS014-treated rats (average weight 425g) had 20% higher body weight than the controls rats (average 360 g). When i.c.v. injections were terminated, the body weight of the twice-daily HS014-treated rats returned to the levels of control group, whereas the rats treated with continuous infusion of HS014 remained hyperphagic and still gained weight. Blood glucose levels in the rats treated with HS014 infusion were significantly increased. Analysis of body composition in HS014-infused rats indicated that body weight was increased due to fat deposits. These data show for the first time that chronic administration of exogenous MC4 receptor antagonist causes hyperphagia and severe obesity in rats. These data also indicate that the melanocortic control of food intake is very robust and suggest that changes induced by such treatment overcome negative feedback signals.


Nitric oxide is involved in the regulation of guard mother cell division by inhibiting the synthesis of ACC.
A stoma forms by a series of asymmetric divisions of stomatal lineage precursor cell and the terminal division of a guard mother cell (GMC). GMC division is restricted to once through genetic regulation mechanisms. Here, we show that nitric oxide (NO) is involved in the regulation of the GMC division. NO donor treatment results in the formation of single guard cells (SGCs). SGCs are also produced in plants that accumulate high NO, whereas clustered guard cells (GCs) appear in plants with low NO accumulation. NO treatment promotes the formation of SGCs in the stomatal signalling mutants sdd1, epf1 epf2, tmm1, erl1 erl2 and er erl1 erl2, reduces the cell number per stomatal cluster in the fama-1 and flp1 myb88, but has no effect on stomatal of cdkb1;1 cyca2;234. Aminocyclopropane-1-carboxylic acid (ACC), a positive regulator of GMC division, reduces the NO-induced SGC formation. Further investigation found NO inhibits ACC synthesis by repressing the expression of several ACC SYNTHASE (ACS) genes, and in turn ACC represses NO accumulation by promoting the expression of HEMOGLOBIN 1 (HB1) encoding a NO scavenger. This work shows NO plays a role in the regulation of GMC division by modulating ACC accumulation in the Arabidopsis cotyledon.


Case Series of Early Structural Valve Deterioration of Trifecta Bioprosthesis - New Zealand Experience.
Structural valve deterioration (SVD) remains a limitation on the use of bioprosthetic valves, with patient and valve-related factors contributing to early SVD. The Trifecta valve has been reported to have excellent hemodynamics but studies have highlighted early failure. We present a review and case series at a New Zealand tertiary hospital defining early SVD as failure within 3 years of implant. A retrospective review from January 2015 to July 2019 included 525 patients undergoing surgical aortic valve replacement with 263 patients receiving an Abbott Trifecta or Trifecta Glide Technology (GT) valve. Our review found an acceptable safety profile for the valve with excellent hemodynamics, with a low mortality, stroke, and permanent pacemaker rate. Three patients out of 263 were identified from the study period as having early SVD requiring reintervention within 3 years of valve implantation leading to a 1.14% failure rate. One of the valves that had early SVD was a new generation Trifecta GT. An additional four patients were identified to have valves implanted prior to the study period and had valve failure at greater than 3 years post implantation. Five cases had cusp tears as their mechanism of failure, raising concerns about durability. The Trifecta valve has an acceptable safety profile and offers good hemodynamics due to the externally mounted leaflets. However, our experience of early SVD and failure is concerning for valve durability. Further comparison to other bioprosthetic valves and longer term follow-up are required to characterize the mechanism of failures.


[Hematologic profile in pregnancy].
We show a study about haematologic profile at 214 pregnant women, who came to prenatal control at Gynecology and Obstetrics service in the Hospital General de México. This study was realized to stablish the haematologic profile and to have better basis for interpret the different changes that ocur during pregnancy.


Mild respiratory symptoms in asthmatic patients might not be due to bronchoconstriction.
Although respiratory symptoms in asthmatic patients are likely to be caused by bronchoconstriction, this should be confirmed by spirometry. In this study, our aim was to determine the percentage of asthmatic patients who present with mild respiratory symptoms but fail to show any evidence of bronchoconstriction in spirometry. A total of 428 known asthmatic patients (57.5% females) participated in the study. Inclusion criteria were age ≥16 years, known asthmatics for at least 1 year, presenting with mild respiratory symptoms including cough, wheezes, shortness of breath, and chest tightness. Patients presenting with moderate or severe asthma exacerbations were excluded from the study. Spirometry measurements were performed according to the guidelines of the American Thoracic Society. SPSS was used for data analysis. The percentage of patients who did not show any evidence of airway obstruction was calculated. For spirometry variables, mean and standard deviation were calculated. For the categorical variables, Chi-square test was performed to determine statistical significance at alpha=0.05. Typical obstructive pattern was found in 38 (or 9%) of all participants. Evidence of obstruction within small or middle airways was found in all those who showed an obstructive pattern and more than 90% of those who showed restrictive or mixed patterns. About 11% of the participants showed a normal spirometric pattern with no evidence of small airway obstruction. Statistical analysis showed an insignificant relation between patterns of spirometry and gender or body mass index of the participants. About 11% of asthmatic patients with mild respiratory symptoms who attended the respiratory clinic have no evidence of bronchoconstriction. Spirometry is an essential step for evaluation of every asthmatic patient who presents with respiratory symptoms.


Volume estimation of excimer laser tissue ablation for correction of spherical myopia and hyperopia.
To determine the theoretical volumes of ablation for the laser treatment of spherical refractive errors in myopia and hyperopia. The cornea was modeled as a spherical shell. The ablation profiles for myopia and hyperopia were based on an established paraxial formula. The theoretical volumes of the ablated corneal lenticules for the correction of myopia and hyperopia were calculated by two methods: (1) mathematical approximation based on a simplified geometric model and (2) finite integration. These results were then compared for optical zone diameters of 0.5 to 11.00 mm and for initial radii of curvature of 7.5, 7.8, and 8.1 mm. Referring to a simplified geometrical model, the volume of ablated corneal tissue was estimated to be proportional to the magnitude of treatment (D) and to the fourth power of the treatment diameter (S(4)). For refractive correction of myopia and hyperopia, volume estimations using our formula, V congruent with D. (S/9)(4), were similar to those obtained by finite integration for optical zone diameters of 0.5 to 8.5 mm and for corneal radii of curvature within the clinical range (7.5, 7.8, and 8.1 mm). The theoretical volume of corneal tissue ablated within the optical zone for spherical corrections can be accurately approximated by this simplified formula. This may be helpful in evaluating factors that contribute to corneal ectasia after LASIK for myopia and hyperopia. Treatment diameter (S) is the most important determinant of the volume of tissue ablation during excimer laser surgery.


Leukotrienes C4 and D4 increase the ciliary beat frequency in human upper airway mucosa in vitro.
It has been suggested that leukotrienes C4 (LTC4) and D4 (LTD4) released from upper respiratory mucosa influence mucociliary transport during allergic reactions. We studied the in vitro effects of leukotrienes C4 and D4 on the ciliary beat frequency (CBF) of human adenoid explants over a 5-hour period. Tissue explants were cultured at 35 degrees C in Minimum Essential Medium Eagle (MEM). The CBF was measured using phase contrast microscopy and microphotometry. Measurements of CBF were recorded in medium alone and in medium containing LTC4 or LTD4 at concentrations of 10(-8) and 10(-6) M. LTC4 and LTD4 increased CBF at concentrations of 10(-8) and 10(-6) M with increases of 20.51% +/- 2.69% and 29.84% +/- 4.06%, respectively. To determine the specificity of the LTC4 and LTD4 effects, the ciliated epithelium was treated with the specific leukotriene receptor antagonist LY-171,883 before administration of LTC4 and LTD4. LY-171,883 (10(-6) M) significantly inhibited the ciliostimulatory effects of both leukotrienes. Our findings indicate that LTC4 and LTD4 increase CBF in vitro by activation of the LTD4 receptor.


Postdischarge growth failure among extremely low birth weight infants: Correlates and consequences.
To describe the prevalence and correlates of postdis-charge growth failure among extremely low birth weight (ELBW) infants (weighing less than 1 kg) and its impact on growth and development. One hundred fifty-four ELBW infants were followed to 20 months corrected age. Growth failure was defined as a decrease in weight z score (standard deviation score) of over 0.67 during one of three periods: between 40 weeks and four months (period I), between four and eight months (period II), and between eight and 20 months corrected age (period III). A decrease in weight z score of this magnitude is comparable to crossing major growth percentiles (eg, from 25th to 10th percentile). Developmental outcomes were assessed at 20 months. Growth failure occurred in 8% of infants during period I, 28% during period II and 12% during period III. This resulted in poorer growth attainment and motor function at 20 months. Significant predictors of growth failure included chronic lung disease during periods I and III, and cerebral palsy during period III. Growth failure during period II, while more common, was not associated with specific sequelae of prematurity. It may represent a physiological process comparable to shifts in weight percentiles in term-born infants. Postdischarge growth failure is common among ELBW infants and contributes to poorer growth outcomes. It is associated with poorer motor outcomes when it occurs early after discharge or later in infancy. A decrease in weight z score of over 0.67 can serve as a useful indicator of growth failure in ELBW infants.


[Novonal (diaethylallylacetamide) poisoning (author's transl)].
Novonal, a hypnotic dispensed without medical prescription in West Germany, has in the past two years been used relatively often in suicide attempts. It is quickly absorbed after oral intake, with a blood half-life of 6-11 hours, most of it being excreted by the kidneys after metabolic degradation. Analysis of 23 cases of Novonal intoxication revealed that cardiac arrhythmias, hypotension and respiratory depression may occur with 6 g Novonal (contained in a pocket of 20 tablets) and must thus be rated as potentially lethal. In severe cases of poisoning haemoperfusion or haemodialysis may be useful. Pharmacological data suggest that forced diuresis is ineffective.


The importance of asepsis in dental implantology.
All surgical procedures, including dental implantation, involve certain risk factors. One of the most common is that of postoperative infection. The harsh realities of today's medical environment such as the threat of blood-borne pathogens and transmittable infections, the increased cost of hospitalization, and the increase in malpractice suites against surgeons, make it imperative that all surgeons follow safe and sterile surgical protocols that reduce the chances of infections and ensure satisfactory results.


Immune evaluation with skin testing. A study of testicular, prostatic, and bladder neoplasms.
Fifty patients with testicular carcinoma, 45 with prostatic neoplasm, 84 with bladder carcinoma, and 13 with benign bladder papilloma were evaluated for skin reactivity to DNCB and other intradermal antigens. Correlation between pathologic staging and skin-test reactivity was sought. Reaction to DNCB among patients with testis tumors was more significantly depressed by chemotherapy than by the extent of retroperitoneal or distant metastatic disease indicating that skin testing as a means of following the course of disease or of predicting survival may be limited by alterations caused by chemotherapy. DNCB reactivity did not correlate with the prognosis for the different stages of disease, but follow-up studies of individual patient survival are needed for substantiation. Depression of DNCB reactivity exists among patients with prostatic carcinoma whether the disease is localized or widely metastatic. Only lengthy follow-up will determine if there is any correlation of reactivity with survival in individual patients. DNCB reactivity among patients with bladder tumors shows progressive reduction with increasing stage disease and lends support to the evidence suggesting immune deficiency in patients with bladder neoplasm.


Robust Bipolar Light Emission and Charge Transport in Symmetric Molecular Junctions.
Molecular junctions consisting of a Ru(bpy)3 oligomer between conducting carbon contacts exhibit an exponential dependence of junction current on molecular layer thickness (d) similar to that observed for other aromatic devices when d < 4 nm. However, when d > 4 nm, a change in transport mechanism occurs which coincides with light emission in the range of 600-900 nm. Unlike light emission from electrochemical cells or solid-state films containing Ru(bpy)3, emission is bipolar, occurs in vacuum, has rapid rise time (<5 ms), and persists for >10 h. Light emission directly indicates simultaneous hole and electron injection and transport, possibly resonant due to the high electric field present (>3 MV/cm). Transport differs fundamentally from previous tunneling and hopping mechanisms and is a clear "molecular signature" relating molecular structure to electronic behavior.


Immediate effect of three different electroacupuncture protocols on fasting blood glucose in obese patients: a pilot study.
Obesity is an increasing global health problem, and current methods of management are limited. Preliminary research data suggest that acupuncture may have an influence on metabolic parameters related to obesity. To determine the electroacupuncture (EA) protocol to be used in a future clinical trial examining the effect of acupuncture on metabolic parameters related to obesity and to examine whether a single EA treatment can change fasting blood glucose in obese subjects. 16 obese women aged 30-52 years with body mass index >30 kg/m(2) were assigned consecutively into three groups and their fasting blood glucose was measured before and after administering a single session, lasting 30 min, of one of three EA treatment protocols. The Dorsal group received EA to dorsal segmental acupuncture points BL18-23 bilaterally (corresponding to the segmental levels innervating the pancreas); the Ear group received EA to ear points in the cavum conchae; and the Limb group received EA to points in the arms and legs (LI10-LI11, ST36-Zongping). After a single session of EA there was a statistically significant decrease in fasting blood glucose in the Dorsal and Limb groups, but there was no change and even a trend towards an increase in the glucose level in the Ear group. The findings of this small pilot study suggest that EA to either dorsal segmental points corresponding to the pancreas or to muscle points in all four limbs may exert a beneficial effect on glucose metabolism in obese women.


Brush border enzyme hydrolysis and glycaemic effects of isomaltulose compared to other saccharides in dogs.
Digestible carbohydrates differ in glycaemic response, therewith having the potential to influence metabolic conditions such as insulin resistance and diabetes mellitus. Isomaltulose has been proven to lower the glycaemic response in humans, which to date has not been studied in dogs. Therefore, the aim of the present study was to characterise the digestibility, as well as the physiological effects of isomaltulose in dogs, in comparison to other saccharides. To this end, three studies were performed. Study 1 was an in vitro study, evaluating the small intestinal hydrolysis of isomaltulose compared to other relevant carbohydrate sources. Three of these saccharides, having close and low-moderate degrees of hydrolysis by brush border enzymes, were also evaluated in vivo for their glycaemic effects by measuring plasma levels of glucose, insulin and glucagon-like peptide 1 (GLP-1) 0-180 min after administration of a single dosage after an overnight fast (i.e., isomaltulose, sucrose and maltodextrin in a 3 × 3 Latin-square design, in 9 dogs, Study 2). To understand if digestive enzymes, underlying glycaemic responses for isomaltulose and sucrose can be upregulated, we exposed dogs to these saccharides for 2 weeks and repeated the measurements after an overnight fast in 18 dogs (Study 3). Isomaltulose was hydrolysed by intestinal enzyme preparation from all three dogs, but the degrading activity was low (e.g., 3.95 ± 1.03 times lower vs. sucrose), indicating a slower rate of hydrolysis. Isomaltulose had a low glycaemic response, in line with in vitro data. In vitro hydrolysis of sucrose was comparable or even higher than maltodextrin in contrast to the more pronounced glycaemic response to maltodextrin observed in vivo. The numerically higher blood glucose response to sucrose after continuous consumption, might indicate an adaptive response. In conclusion, the current work provides valuable insights into the digestion physiology of various saccharides in dogs. Further investigations on related benefits are thus warranted.


Multiplex Protein Profiling by Low-Signal-Loss Single-Cell Western Blotting with Fluorescent-Quenching Aptamers.
Single-cell western blotting (scWB) is a prevalent technique for high-resolution protein analysis on low-abundance cell samples. However, the extensive signal loss during repeated antibody stripping precludes multiplex protein detection. Herein, we introduce Fluorescent-quenching Aptamer-based Single-cell Western Blotting (FAS-WB) for multiplex protein detection at single-cell resolution. The minimal size of aptamer probes allows rapid in-gel penetration, diffusion, and elution. Meanwhile, the fluorophore-tagged aptamers, coordinated with complementary quenching strands, avoid the massive signal loss conventionally caused by antibody stripping during repeated staining. Such a strategy also facilitates multiplex protein analysis with a limited number of fluorescent tags. We demonstrated FAS-WB for co-imaging four biomarker proteins (EpCAM, PTK7, HER2, CA125) at single-cell resolution with lower signal loss and enhanced signal-to-noise ratio compared to conventional antibody-based scWB. Being more time-saving (less than 25 min per cycle) and economical (1/1000 cost of conventional antibody probes), FAS-WB offers a highly efficient platform for profiling multiplex proteins at single-cell resolution.


Effects of sleep pressure on endogenous cardiac autonomic activity and body temperature.
This study investigated the effects of variations in sleep pressure on cardiac autonomic activity and body temperature. In a counterbalanced design, 12 healthy, young subjects (6 men and 6 women) remained recumbent during 30 h of wakefulness (high sleep pressure) and 6 h of wakefulness (low sleep pressure). Both periods of wakefulness were immediately followed by a sleep opportunity, and the first 2 h of sleep were analyzed. During extended hours of wakefulness, a reduction in heart rate was mediated by a decline in cardiac sympathetic activity (measured via preejection period) and the maintenance of cardiac parasympathetic activity (measured via respiratory sinus arrhythmia). In subsequent high-pressure sleep, parasympathetic activity was amplified and sympathetic activity was negatively associated with electroencephalographic slow-wave activity. Sleep deprivation had no impact on foot temperature, but it did alter the pattern of change in core body temperature. A downregulation of cardiac autonomic activity during both extended hours of wakefulness and subsequent sleep may respectively provide "protection" and "recovery" from the temporal extension of cardiac demand.


Key principles involved in applying and removing wound dressings.
This article aims to help practitioners consider a variety of wound dressings and develop their knowledge about the use of dressings to ensure patients are protected and care is cost-effective. The article focuses on which dressing should be used depending on wound type and how different dressings should be applied and removed. Author


[Molecular characterization of O-antigen gene cluster of Escherichia coli O23 reference strain and identification of UDP-N-acetylglucosamine 4-epimerase].
Lipopolysaccharide (LPS) is one of the major components of the outer membrane of gram-negative bacteria. It is an amphipathic molecule composing of lipid A, a core oligosaccharide and an O-specific antigen. O-antigen, which is a repeat unit polysaccharide, is a major contribution to the antigenic variability of the bacterial cell surface. Genes involved in O-antigen biosynthesis are generally found to be clustered between the housekeeping genes galF and gnd on the chromosome of E. coli. E. coli O23 is one of the enterotoxigenic E. coli causing pediatric diarrhea in the developing world. The O-antigen gene cluster of E. coli O23 type strain was amplified by long-range PCR using primers based on galF and gnd and then sequenced. Except for galF and gnd, seven open reading frames were identified and assigned functions on the basis of their similarity to those from available databases. The seven genes include a UDP-N-acetylglucosamine 4-epimerase gene (gne), the O-antigen polymerase gene (wzy), the O-antigen transferase gene (wzx) and four glycosyltransferase genes (orf2, orf4, orf5, orf6). The UDP-N-acetylglucosamine 4-epimerase (Gne) was identified by mutation and complementation complement tests. The structure of Gne was predicted by the homology modeling method, and the active sites were also analyzed. The phylogenetic and structural analysis showed that the Gne derived from the common ancestor with E. coli O23 Gne were UDP-GlcNAc/UDP-GalNAc epimerases. The specific DNA used for rapid molecular genotyping for E. coli O23 was also identified.


Modeling the dynamics of transcriptional gene regulatory networks for animal development.
The dynamic process of cell fate specification is regulated by networks of regulatory genes. The architecture of the network defines the temporal order of specification events. To understand the dynamic control of the developmental process, the kinetics of mRNA and protein synthesis and the response of the cis-regulatory modules to transcription factor concentration must be considered. Here we review mathematical models for mRNA and protein synthesis kinetics which are based on experimental measurements of the rates of the relevant processes. The model comprises the response functions of cis-regulatory modules to their transcription factor inputs, by incorporating binding site occupancy and its dependence on biologically measurable quantities. We use this model to simulate gene expression, to distinguish between cis-regulatory execution of "AND" and "OR" logic functions, rationalize the oscillatory behavior of certain transcriptional auto-repressors and to show how linked subcircuits can be dealt with. Model simulations display the effects of mutation of binding sites, or perturbation of upstream gene expression. The model is a generally useful tool for understanding gene regulation and the dynamics of cell fate specification.


Dosing parameters for the effects of high-frequency transcranial magnetic stimulation on smoking cessation: study protocol for a randomized factorial sham-controlled clinical trial.
Despite the considerable success of comprehensive tobacco control efforts, tobacco use remains one of the greatest preventable causes of death and disease today. Over half of all smokers in the US make quit attempts every year, but over 90% relapse within 12 months, choosing the immediate reinforcement of smoking over the long-term benefits of quitting. Conceptual and empirical evidence supports continued investigation of high frequency repetitive transcranial magnetic stimulation (rTMS) of the left dorsolateral prefrontal cortex in reducing relapse and decreasing cigarette consumption. While this evidence is compelling, an optimal dosing strategy must be determined before a long-term efficacy trial can be conducted. The goal of this study is to determine a dosing strategy for 20 Hz rTMS that will produce the best long-term abstinence outcomes with the fewest undesirable effects. This is a fully crossed, double-blinded, sham-controlled, 3x2x2 randomized factorial study. The three factors are duration (stimulation days: 8, 12, and 16); intensity (900 or 1800 pulses per day); and sham control. Participants (n = 258) will consist of adults (18-65) who are motivated to quit smoking cigarettes and who will be followed for 6 months post-quit. Outcomes include latency to relapse, point prevalence abstinence rates, delay discounting rates, cognitive-behavioral skills acquisition, and multiple measures of potential undesirable effects that impact participant compliance. This study integrates existing theoretical concepts and methodologies from neuropsychology, behavioral economics, brain stimulation, clinical psychology, and the evidence-based treatment of tobacco dependence in the development of a promising and innovative approach to treat tobacco dependence. This study will establish an optimal dosing regimen for efficacy testing. Findings are expected to have a significant influence on advancing this approach as well as informing future research on clinical approaches that combine rTMS with other evidence-based treatments for tobacco dependence and perhaps other addictions. Clinical Trials NCT03865472 (retrospectively registered). The first participant was fully enrolled on November 26, 2018. Registration was posted on March 7, 2019.


p53-like protein binding affinity to the hormone responsive element of the haptoglobin gene in fetal rat livers.
In order to identify nucleoproteins involved in transcriptional regulation of the haptoglobin (Hp) gene, fetal rat livers of dams exposed to inflammation on day 19 of pregnancy were used. Previously observed acute phase-dependent elevation of Hp gene transcriptional activity in prenatal liver was accompanied by increased binding affinities of several fetal soluble nucleoproteins and the hormone response element (RE) of the Hp gene (-170/-56). One of these proteins, a hepatic nucleoprotein of 53 kDa, was identified by Western blotting analysis as a protein within the same molecular mass and epitopes as transcription factor p53. Also, in vitro phosphorylation experiments revealed that the examined fetal nucleoprotein could be liable to the same phosphorylative post-translational modification as p53. The obtained results suggest that the fetal 53 kDa-nucleoprotein could be a homologue of transcription factor p53, participating in the transcriptional modulation of the Hp gene throughout prenatal hepatic development.


Measurement method for slant visibility with slant path scattered radiance correction by lidar and the SBDART model.
Different from the existing methods for estimating averaged slant visibility by lidar and the traditional Koschmieder visibility formula, a measurement method for slant visibility is fundamentally proposed in this paper that considers the correction of slant path scattered radiance. Lidar is adopted to provide aerosol parameters, including optical depth and scattering parameters, and the SBDART (Santa Barbara DISORT Atmospheric Radiative Transfer) model is used to solve the radiative transfer equation to obtain the corresponding radiance distribution; thus, the corrected apparent brightness contrast between the object and background along the slant path is used to achieve accurate slant visibility. Based on the measurement principle of slant visibility, a theoretical simulation and an analysis of the slant path scattered radiance are performed, and the resulting slant visibility is studied in detail in this paper.


Transcriptome analysis of Ruditapes philippinarum hepatopancreas provides insights into immune signaling pathways under Vibrio anguillarum infection.
The Manila clam, Ruditapes philippinarum, is one of the most economically important aquatic clams that are harvested on a large scale by the mariculture industry in China. However, increasing reports of bacterial pathogenic diseases have had a negative effect on the aquaculture industry of R. philippinarum. In the present study, the two transcriptome libraries of untreated (termed H) and challenged Vibrio anguillarum (termed HV) hepatopancreas were constructed and sequenced from Manila clam using an Illumina-based paired-end sequencing platform. In total, 75,302,886 and 66,578,976 high-quality clean reads were assembled from 101,080,746 and 99,673,538 raw data points from the two transcriptome libraries described above, respectively. Furthermore, 156,116 unigenes were generated from 210,685 transcripts, with an N50 length of 1125 bp, and from the annotated SwissProt, NR, NT, KO, GO, KOG and KEGG databases. Moreover, a total of 4071 differentially expressed unigenes (HV vs H) were detected, including 903 up-regulated and 3168 down-regulated genes. Among these differentially expressed unigenes, 226 unigenes were annotated using KEGG annotation in 16 immune-related signaling pathways, including Toll-like receptor, NF-kappa B, MAPK, NOD-like receptor, RIG-I-like receptor, and the TNF and chemokine signaling pathways. Finally, 20,341 simple sequence repeats (SSRs) and 214,430 potential single nucleotide polymorphisms (SNPs) were detected from the H and HV transcriptome libraries. In conclusion, these studies identified many candidate immune-related genes and signaling pathways and conducted a comparative analysis of the differentially expressed unigenes from Manila clam hepatopancreas in response to V. anguillarum stimulation. These data laid the foundation for studying the innate immune systems and defense mechanisms in R. philippinarum.


[The genetic bases of neurodevelopmental disorders].
In the last decade, progress made in genetics is questioning the current implicit nosological model in the Diagnostic and Statistical Manual of Mental Disorders, fourth edition, text revision (DSM-IV-TR) and the International Classification of Diseases, tenth revision. Both the categorical nature and the comorbidity detected on applying diagnostic criteria become unsustainable in the light of the genetic architecture that is emerging from studies being conducted on the genetics of mental disorders. The classical paradigms -one gene for one disease- or even a specific distinctive genetic pattern for each condition, are concepts restricted to specific cases. In this review the objective is to describe the current scenario that has arisen following the latest advances in genetics. The lines of work being traced by research both in the present and in the near future include: the identification of variations in the number of copies (both frequent and rare), indiscriminately linked to different disorders; the concurrence of multiple variants for a single disorder; the double hit phenomenon; and epigenetic modulation. The new version of the DSM, fully aware of the deficiencies in the current model, will mark a turning point that, while somewhat timid, is decidedly oriented towards incorporating a dimensional conception of mental disorders.


Report of the National Institutes of Health (NIH) Workshop on the Development of Research Priorities in Eating Disorders.
The National Institutes of Health (NIH) Workshop on the Development of Research Priorities in Eating Disorders was convened in New York on April 24 and 25, 1996. The goals of the workshop were (1) to identify important unanswered questions in the study and treatment of eating disorders, (2) to discuss potentially fruitful approaches to answering these questions through basic and clinical research, and (3) to assist the NIH and other funding agencies in assigning priorities for research on eating disorders. The program consisted of a series of brief presentations by moderators, each followed by facilitated discussion of the topic with members of the audience. Three reporters (CMG, MJD, FMC) took detailed notes of the proceedings, which have been incorporated into this article. A summary of this workshop is presented, along with recommendations for future research that were identified by workshop participants.


Acute effects of nisoldipine, propranolol, and their combination in patients with chronic stable angina: a double-blind, randomized, cross-over, placebo-controlled study.
We studied the short-term effects of oral administration of nisoldipine (10 mg) and propranolol (80 mg) alone and in combination in 14 patients with chronic exertional angina pectoris in a double-blind, randomized, cross-over study. The 14 patients (13 men and 1 woman, mean age 56 +/- 7 years) performed symptoms-limited bicycle exercise stress test 3 h after placebo or active substance administration. Maximal work load, exercise duration, and time to 1-mm ST segment depression were significantly increased and ST depression at peak exercise was significantly decreased by drugs alone and in combination. Propranolol and nisoldipine alone improved exercise duration similarly and as well as the combination; however, a different response to the three pharmacologic interventions was found in patients treated with single drugs. The improvement in exercise tolerance was associated with rate-pressure product values at peak exercise, unchanged after nisoldipine and significantly reduced after both propranolol alone and in combination. After placebo, all patients had exercise-induced angina, in 9, 8, and 4 patients after nisoldipine, propranolol, and the combination of the two drugs, respectively. Nisoldipine is effective in the treatment of effort angina and its combination with propranolol may be useful and superior in patients who show poor response to monotherapy.


Prevalence of Psychological Stress Among Women Attending the Assisted Reproductive Technology (ART) Unit at King Abdulaziz Medical City: A COVID-19 Pandemic Experience.
Aim The present study aimed to measure the prevalence of psychological stress among women attending the Assisted Reproductive Technology (ART) unit in King Abdulaziz Medical City during the COVID-19 pandemic. Method The study adopted a descriptive cross-sectional design and was conducted between January 2020 and December 2022. A sample of 104 women attending the ART unit were recruited in this study. To collect data, the researchers developed a study questionnaire that consisted of 26 items. Nine items were designed to measure the participants' demographic characteristics, and 17 items were designed to depict their responses related to psychological stress during the COVID-19 pandemic. Data were analyzed using the Statistical Product and Service Solutions (SPSS) (v. 26l; IBM SPSS Statistics for Windows, Armonk, NY). Chi-square and t-tests were used to assess the association between stress and sociodemographics. Results The findings of the study showed an overall prevalence of 86.53% (n=90). Levels of psychological stress were moderate (34.62%, n=36), severe (39.42%, n=41), and extremely severe (12.50%, n=13). The study found that there is a significant statistical interaction between the number of previous ART attempts and psychological stress (p=0.0019). Conclusion The study concluded that women attending the ART unit at King Abdulaziz Medical City (KAMC), Riyadh, Saudi Arabia, were experiencing high levels of psychological stress, and these levels were affected by the number of previous ART attempts.


HIV-1 viral load measurement in venous blood and fingerprick blood using Abbott RealTime HIV-1 DBS assay.
HIV RNA suppression is a key indicator for monitoring success of antiretroviral therapy. From a logistical perspective, viral load (VL) testing using Dried Blood Spots (DBS) is a promising alternative to plasma based VL testing in resource-limited settings. To evaluate the analytical and clinical performance of the Abbott RealTime HIV-1 assay using a fully automated one-spot DBS sample protocol. Limit of detection (LOD), linearity, lower limit of quantitation (LLQ), upper limit of quantitation (ULQ), and precision were determined using serial dilutions of HIV-1 Virology Quality Assurance stock (VQA Rush University), or HIV-1-containing armored RNA, made in venous blood. To evaluate correlation, bias, and agreement, 497 HIV-1 positive adult clinical samples were collected from Ivory Coast, Uganda and South Africa. For each HIV-1 participant, DBS-fingerprick, DBS-venous and plasma sample results were compared. Correlation and bias values were obtained. The sensitivity and specificity were analyzed at a threshold of 1000 HIV-1 copies/mL generated using the standard plasma protocol. The Abbott HIV-1 DBS protocol had an LOD of 839 copies/mL, a linear range from 500 to 1×107 copies/mL, an LLQ of 839 copies/mL, a ULQ of 1×107 copies/mL, and an inter-assay SD of ≤0.30 log copies/mL for all tested levels within this range. With clinical samples, the correlation coefficient (r value) was 0.896 between DBS-fingerprick and plasma and 0.901 between DBS-venous and plasma, and the bias was -0.07 log copies/mL between DBS-fingerprick and plasma and -0.02 log copies/mL between DBS-venous and plasma. The sensitivity of DBS-fingerprick and DBS-venous was 93%, while the specificity of both DBS methods was 95%. The results demonstrated that the Abbott RealTime HIV-1 assay with DBS sample protocol is highly sensitive, specific and precise across a wide dynamic range and correlates well with plasma values. The Abbott RealTime HIV-1 assay with DBS sample protocol provides an alternative sample collection and transfer option in resource-limited settings and expands the utility of a viral load test to monitor HIV-1 ART treatment for infected patients.


Application of a patient-derived xenograft model in cytolytic viral activation therapy for nasopharyngeal carcinoma.
Nasopharyngeal carcinoma (NPC) is an Epstein Barr virus (EBV)-related malignancy in which the tumor microenvironment plays a pivotal role in tumor progression. Here, we developed two patient-derived xenograft (PDX) mouse lines from engrafted NPC metastatic tumors. Positive staining for EBV-encoded small RNAs confirmed that these tumors harbored EBV, and gene expression profile analyses further showed that the PDX was highly similar to the primary parent tumor. In vivo drug screening using the PDX system demonstrated that gemcitabine had the best antitumor effect among the tested drugs. The donor of this PDX also showed excellent responsiveness to gemcitabine treatment. The combination of gemcitabine and valproic acid exerted synergistic antitumor effects. Further addition of ganciclovir to this two-drug combination regimen enhanced cytolytic viral activation, yielding the best antitumor response among tested regimens. Treatment with this three-drug combination regimen decreased plasma EBV-DNA load, tumor viral concentration, and the number of viable tumor cells to a greater extent than the two-drug gemcitabine and valproic acid combination. These results highlight the value of PDX models in the development of EBV-targeted strategies to treat NPC.


PFO Closure for Cryptogenic Stroke: A Review and Clinical Treatment Algorithm.
With a high prevalence in the general population of approximately 25%, and a prevalence in the cryptogenic stroke population approaching 40%, the propensity of a patent foramen ovale (PFO) to precipitate or enable stroke, especially in young, otherwise healthy individuals, has been the subject of much debate. With proof of concept achieved via imaging modalities documenting thrombus-in-transit, and the development of minimally-invasive percutaneous approaches to closure, multiple observational studies and, more recently, several completed randomized controlled trials have sought to answer the question of when and in whom PFO closure should occur. We describe the historical context of PFO closure and review the observational and randomized control trial evidence in this field, culminating in the recent Food and Drug Administration approval of the first dedicated closure device for PFO. Guidelines and consensus statements are discussed, and a novel treatment algorithm is proposed. Future directions in PFO closure will include new devices, further data from completed and upcoming clinical trials, and potential expansion into other disease states associated with PFO.


Treatment algorithms of patients with impaction of maxillary central incisors caused by supernumerary teeth.
Introduction: Supernumerary teeth, like etiological factor of impaction, occur in 7,05% of patients, are usually located in anterior maxillary region - 93,2%. The aim of the study is to develop an algorithm of treatment of patients with impaction of permanent maxillary central incisors, caused by supernumerary teeth, during periods of mixed and permanent dentition by processing our own approaches to treatment. Materials and methods: Treatment results of seven patients (6, 7 (2 patients), 8, 9, 10 and 15 years old) with delay in eruption of maxillary central incisors were analyzed. According to the results of clinical and additional examination methods (photometric examination of face, study of diagnostic models of jaws and orthopantomograms, 3D computed tomography), impacted central incisors, supernumerary teeth, blocking their eruption, were diagnosed. Results: Clinical experience of treatment of patients with impacted teeth caused by presence of supernumerary teeth in the jaws, allowed us to propose algorithms of providing complex treatment in such cases, depending on physiological state of occlusion - mixed or permanent, and depth of supernumerary tooth location. Conclusions: If there are no central incisors in the dental arch after the term of their physiological eruption, orthopantomogram must be taken to confirms or reject tooth impaction, and then 3D computed tomography should be done to show accurate localization of the tooth and only after these stages it is possible to choose the treatment plan. Patients with impacted maxillary central incisors caused by supernumerary teeth should undergo orthodontic treatment in several stages.


[A cholesterol reducing diet is not useful].
In daily practice a cholesterol-lowering diet with intensive monitoring by dieticians leads to a mean decrease of the serum cholesterol level by 2%. Once the monitoring is discontinued, the serum cholesterol level as a rule returns to baseline. Were it possible to maintain a 5% cholesterol reduction throughout life, occurrence of coronary heart disease would in theory be postponed by one to a few months. In diet studies, with observation lasting from 6 to 15 years, no clear favourable effect on the development of coronary heart disease was demonstrated. Prescription of a cholesterol-reducing diet and referral to a dietician are therefore fairly useless.


All mixed up: defining roles for β-cell subtypes in mature islets.
Following differentiation during fetal development, β cells further adapt to their postnatal role through functional maturation. While adult islets are thought to contain functionally mature β cells, recent analyses of transgenic rodent and human pancreata reveal a number of novel heterogeneity markers in mammalian β cells. The marked heterogeneity long after maturation raises the prospect that diverse populations harbor distinct roles aside from glucose-stimulated insulin secretion. In this review, we outline our current understanding of the β-cell maturation process, emphasize recent literature on novel heterogeneity markers, and offer perspectives on reconciling the findings from these two areas.


[Renal angiomyolipoma: diagnostic difficulties and surgical indications in 20 surgically treated patients].
Between 1982 and 1993, 6 male and 14 female patients underwent partial or total nephrectomy due to renal angiomyolipoma (AML). Presentation forms were: in 6 cases (30%) acute pain with severe haematuria or signs of visceral bleeding, 3 (15%) acute pain without other symptoms, 7 (35%) chronic pain associated to haematuria or fever, or isolated haematuria, and in 4 (20%) it was a chance finding. No significant differences were found among tumor sizes considering the severity of the presentation form, but all AMLs with serious signs and symptoms had more than 5 cm diameter. A pre-operative diagnosis of AML was made in 15 (75%) patients, while 5 (25%) underwent surgery for other presumed diagnosis. Nine were conservative procedures of renal parenchyma and 11 were total or radical nephrectomies. There were no significant differences between tumour sizes by treatment performed. The authors review the imaging diagnostic difficulties and the various criteria supporting surgery in AML, and conclude by recommending individualized diagnosis and treatment in patients with AML suspicion of including puncture biopsy or cytology when in doubt, trying to avoid surgery in well characterized tumours, under 4 or 5 cm, provided that they could be followed-up regularly.


Muscarinic cholinergic receptors on skin fibroblasts in familial affective disorder.
Cultured human skin fibroblasts possess muscarinic receptors with the properties of specific binding, saturability, pharmacologic specificity, inhibition of norepinephrine-stimulated adenylate cyclase, and increased binding after incubation with an antagonist. The number of binding sites appears to be a stable characteristic of each cell line. We studied fibroblasts from 18 patients with a major affective disorder and found that they had a higher density of binding sites than cells from 12 normal controls. Fibroblasts from 18 relatives who had histories of major or minor affective disorder also had a higher density, and those from five normal relatives were similar to controls. These results, although still preliminary, suggest that increased cholinergic-receptor density may be associated with vulnerability to affective disorders in some familial cases.


A transient decrease in reactive oxygen species in roots leads to root hair deformation in the legume-rhizobia symbiosis.
* A possible role for reactive oxygen species (ROS) in root hair deformation in response to Nod factor (NF) was investigated using Medicago truncatula nodulation mutants, and an inhibitor and precursors of ROS. * In wild-type roots, ROS efflux transiently decreased approximately 1 h after NF treatment. Transcript accumulation of two NADPH oxidase homologs, respiratory burst oxidase homolog 2 (MtRBOH2) and MtRBOH3, also transiently decreased at 1 h. However, in the nonnodulating mutant Nod factor perception (nfp), transcript accumulation did not change. * Exogenous application of ROS prevented root hair swelling and branching induced by NF. When accumulation of ROS was prevented by diphenylene iodonium (DPI), NF did not induce root hair branching. Root treatment with DPI alone reduced ROS efflux and induced root hair tip swelling. Transient treatment of roots with DPI mimicked NF treatment and resulted in root hair branching in the absence of NF. A transient DPI treatment did not induce root hair branching in the nonlegumes Arabidopsis thaliana and tomato (Lycopersicon esculentum). * The results suggest a role for the transient reduction of ROS accumulation in governing NF-induced root hair deformation in legumes.


Regulation of Autophagy through TORC1 and mTORC1.
Autophagy is an intracellular protein-degradation process that is conserved across eukaryotes including yeast and humans. Under nutrient starvation conditions, intracellular proteins are transported to lysosomes and vacuoles via membranous structures known as autophagosomes, and are degraded. The various steps of autophagy are regulated by the target of rapamycin complex 1 (TORC1/mTORC1). In this review, a history of this regulation and recent advances in such regulation both in yeast and mammals will be discussed. Recently, the mechanism of autophagy initiation in yeast has been deduced. The autophagy-related gene 13 (Atg13) and the unc-51 like autophagy activating kinase 1 (Ulk1) are the most crucial substrates of TORC1 in autophagy, and by its dephosphorylation, autophagosome formation is initiated. Phosphorylation/dephosphorylation of Atg13 is regulated spatially inside the cell. Another TORC1-dependent regulation lies in the expression of autophagy genes and vacuolar/lysosomal hydrolases. Several transcriptional and post-transcriptional regulations are controlled by TORC1, which affects autophagy activity in yeast and mammals.


Blood Transfusion and Survival for Resected Adrenocortical Carcinoma: A Study from the United States Adrenocortical Carcinoma Group.
Perioperative blood transfusion is associated with decreased survival in pancreatic, gastric, and liver cancer. The effect of transfusion in adrenocortical carcinoma (ACC) has not been studied. Patients with available transfusion data undergoing curative-intent resection of ACC from 1993 to 2014 at 13 institutions comprising the United States Adrenocortical Carcinoma Group were included. Factors associated with blood transfusion were determined. Primary and secondary end points were recurrence-free survival (RFS) and overall survival (OS), respectively. Out of 265 patients, 149 were included for analysis. Out of these, 57 patients (38.3%) received perioperative transfusions. Compared to nontransfused patients, transfused patients more commonly had stage 4 disease (46% vs 24%, P = 0.01), larger tumors (15.8 vs 10.2 cm, P < 0.001), inferior vena cava involvement (24.6% vs 5.4%, P = 0.002), additional organ resection (78.9% vs 36.3%, P < 0.001), and major complications (29% vs 2%, P < 0.001). Transfusion was associated with decreased RFS (8.9 vs 24.7 months, P = 0.006) and OS (22.8 vs 91.0 months, P < 0.001). On univariate Cox regression, transfusion, stage IV, hormonal hypersecretion, and adjuvant therapy were associated with decreased RFS. On multivariable analysis, only transfusion [hazard ratio (HR) = 1.7, 95% confidence interval (CI) =1.0-2.9, P = 0.04], stage IV (HR = 3.2, 95% CI = 1.7-5.9, P < 0.001), and hormonal hypersecretion (HR = 2.6, 95% CI = 1.5-4.2, P < 0.001) were associated with worse RFS. When applying this model to OS, similar associations were seen (transfusion HR = 2.0, 95% CI = 1.1-3.8, P = 0.02; stage 4 HR = 6.2, 95% CI = 3.1-12.4, P < 0.001; hormonal hypersecretion HR = 3.5, 95% CI = 1.9-6.4, P < 0.001). There was no difference in outcomes between patients who received 1 to 2 units versus >2 units of packed red blood cells in median RFS (8.9 vs 8.4 months, P = 0.95) or OS (26.5 vs 18.6 months, P = 0.63). Perioperative transfusion is associated with earlier recurrence and decreased survival after curative-intent resection of ACC. Strategies and protocols to minimize blood transfusion should be developed and followed.


Activation of T and B lymphocytes in vitro. I. Regulatory influence of bacterial lipopolysaccharide (LPS) on specific T-cell helper function.
The present studies were undertaken to analyze the nature of the effect of bacterial lipopolysaccharide (LPS) on antibody production in vitro. We have done this by making comparative studies of the effects of LPS on in vitro primary and secondary antibody responses to soluble hapten-protein conjugates and to particulate and soluble sheep erythrocyte antigens. The results obtained demonstrate that the biological action of LPS in vitro may be predominantly manifested on the function of B lymphocytes or T lymphocytes depending on the conditions employed. In the absence of antigen, LPS appears to act primarily on B lymphocytes. In the presence of antigen, however, the data presented here show that LPS significantly influences specific helper T-cell function and it is this latter influence that is predominantly responsible for the adjuvant effects of LPS on antigen-specific antibody responses.


Function of a mutant ryanodine receptor (T4709M) linked to congenital myopathy.
Physiological muscle contraction requires an intact ligand gating mechanism of the ryanodine receptor 1 (RyR1), the Ca2+-release channel of the sarcoplasmic reticulum. Some mutations impair the gating and thus cause muscle disease. The RyR1 mutation T4706M is linked to a myopathy characterized by muscle weakness. Although, low expression of the T4706M RyR1 protein can explain in part the symptoms, little is known about the function RyR1 channels with this mutation. In order to learn whether this mutation alters channel function in a manner that can account for the observed symptoms, we examined RyR1 channels isolated from mice homozygous for the T4709M (TM) mutation at the single channel level. Ligands, including Ca2+, ATP, Mg2+ and the RyR inhibitor dantrolene were tested. The full conductance of the TM channel was the same as that of wild type (wt) channels and a population of partial open (subconductive) states were not observed. However, two unique sub-populations of TM RyRs were identified. One half of the TM channels exhibited high open probability at low (100 nM) and high (50 μM) cytoplasmic [Ca2+], resulting in Ca2+-insensitive, constitutively high Po channels. The rest of the TM channels exhibited significantly lower activity within the physiologically relevant range of cytoplasmic [Ca2+], compared to wt. TM channels retained normal Mg2+ block, modulation by ATP, and inhibition by dantrolene. Together, these results suggest that the TM mutation results in a combination of primary and secondary RyR1 dysfunctions that contribute to disease pathogenesis.


N-acetylaspartate (NAA) correlates inversely with cannabis use in a frontal language processing region of neocortex in MDMA (Ecstasy) polydrug users: a 3 T magnetic resonance spectroscopy study.
Impaired verbal memory is common in MDMA (Ecstasy) polydrug users. The contributions of Ecstasy or polydrug exposure to reduced verbal memory are unclear, as is the neural basis for this cognitive deficit. Ecstasy users have reduced gray matter in brain regions mediating verbal memory (BA 18, 21 and 45). N-acetylaspartate (NAA) as a neuronal marker and myoinositol (mI) as a glial marker are inconsistently affected in Ecstasy users. We used 3 T MRS in 17 recreational drug users to test the hypothesis that Ecstasy polydrug use would be associated with altered NAA or mI in BA 18, 21 and 45. No effects were seen for mI. Metabolite ratios for NAA (mean+/-SD) were: BA 18-NAA/Cr (2.030+/-0.188); BA 21-NAA/Cr (1.861+/-0.325); BA 45-NAA/Cr (1.925+/-0.329). Lifetime cannabis use was significantly associated with BA 45 NAA/Cr (r=-0.687, p=0.014) but not with NAA in BA 18 or 21. In contrast, there were no statistically significant associations for lifetime use of Ecstasy, alcohol, or cocaine with NAA. These findings suggest that cannabis use may contribute to altered neuronal integrity in Ecstasy polydrug users in a brain region associated with verbal memory processing.


Exact conditional distribution of a three-phase invariant in space group P1. IV. Further improvements of Cochran-like approximations.
A new approximate method of computing the conditional probability density function (c.p.d.f.) of a three-phase invariant is investigated, the results being compared to accurate calculations [Shmueli, Rabinovich & Weiss (1989). Acta Cryst. A45, 361-367]. A direct dependence on N, the number of atoms in the unit cell, is incorporated into the new approximation and its performance in the equal-atom case appears to be excellent over a large range of N values. The polynomial approximation of the recently published method [Posner, Shmueli & Weiss (1993). Acta Cryst. A49, 260-265] has been improved in both accuracy and range.


Par6 is phosphorylated by aPKC to facilitate EMT.
The conserved polarity proteins Par6 and aPKC regulate cell polarization processes. However, increasing evidence also suggests that they play a role in oncogenic progression. During tumor progression, epithelial to mesenchymal transition (EMT) delineates an evolutionary conserved process that converts stationary epithelial cells into mesenchymal cells, which have an acquired ability for independent migration and invasion. In addition to signaling pathways that alter genetic programes that trigger the loss of cell-cell adhesion, alternative pathways can alter cell plasticity to regulate cell-cell cohesion and increase invasive potential. One such pathway involves TGFβ-induced phosphorylation of Par6. In epithelial cells, Par6 phosphorylation results in the dissolution of junctional complexes, cytoskeletal remodelling, and increased metastatic potential. Recently, we found that aPKC can also phosphorylate Par6 to drive EMT and increase the migratory potential of non-small cell lung cancer cells. This result has implications with respect to homeostatic and developmental processes involving polarization, and also with respect to cancer progression-particularly since aPKC has been reported to be an oncogenic regulator in various tumor cells.


Pharmacological features of vascular responses of isolated dog and monkey lingual arteries to vasoactive substances.
Using a perfusion technique of isolated vessels, vasoconstrictor responses to alpha-adrenoceptor agonists (norepinephrine [NE], phenylephrine [PE], clonidine, xylazine and tyramine) and KCl were investigated in isolated, perfused dog and monkey lingual arteries. A stainless steel cannula was inserted into the lingual artery segment and perfused with Krebs-Henseleit solution at a constant flow rate. In dog lingual arteries, the agonists induced vasoconstrictions with the following order of potency: NE greater than PE greater than tyramine much greater than clonidine greater than xylazine greater than KCl. In monkey preparations, the order was NE greater than PE much greater than clonidine greater than or equal to tyramine greater than xylazine greater than KCl. In both preparations, NE- and PE-induced constrictions were blocked by bunazosin (an alpha-1 adrenoceptor antagonist), but not influenced by midaglizole (a potent alpha-2 antagonist). Diltiazem (a Ca entry blocker) significantly attenuated NE-induced vasoconstrictions in dog lingual arteries, but did not significantly influence these in monkey preparations. These results suggest that: [1] these arteries contain mostly alpha-1 but scarcely any alpha-2 adrenoceptors; [2] in dog preparations, tyramine induced a marked vasoconstriction which may contribute to investigation on the mechanisms of catecholamine releases from sympathetic nerve terminals; and [3] different blocking effects of diltiazem may indicate that extracellular Ca++ influx may have varying degrees of importance in alpha-1 adrenoreceptor-mediated constrictions in different species, although participation of an intracellular mechanism might not be ruled out.


Dichlorodiphenyltrichloroethane (DDT) induced extracellular vesicle formation: a potential role in organochlorine increased risk of Parkinson's disease.
A number of studies have demonstrated that rural living and exposure to pesticides such as dichlorodiphenyltrichloroethane (DDT) highly increase the chances of developing Parkinson's disease. In a previous work, we have found that DDT leads to the formation of vesicular buds that are released from the cells upon fusion of an intermediate endocytic compartment with the plasma membrane. Since extracellular vesicles like exosomes have been implicated in the development of neurodegenerative diseases through the propagation of neurotoxic misfolded proteins from neuron to neuron, in this minireview we propose that organochlorine pesticides could enhance the risk of neurodegenerative diseases by increasing the formation of exosomes.


Effect of glucocorticoid-induced osteoporotic-like conditions on osteoblast cell attachment to implant surface microtopographies.
The objectives of this work were to: (1) establish methodology for pretreating osteoblast-like cells in vitro with dexamethasone to cause glucocorticoid-induced osteoporosis, (2) perform quantitative and qualitative assessments of cellular attachment of osteoporosis-like osteoblasts when grown on implant surfaces of differing roughness, (3) and explore the hypothesis that dexamethasone-treated osteoblasts have altered cell attachment properties by focal adhesion disassembly and decreased tyrosine phosphorylation of the focal adhesion tyrosine kinase. Osteoblasts were cultured with dexamethasone (10(-7) and 10(-6) M) for up to 4 days of incubation to induce osteoporosis-like conditions. Cellular attachment assays demonstrated the effect of dexamethasone treatments on cellular attachment properties of osteoblasts. Qualitative data were obtained utilizing immunofluorescent microscopy and Western blotting. Focal adhesion kinase (FAK) immunoprecipitation and tyrosine-phosphorylation Western blots were obtained from dexamethasone-treated human embryonic palatal mesenchymal- 1486 osteoblast cultures supplemented with ascorbate and beta-glycerol phosphate medium. Cellular attachment was significantly greater (P < 0.05) with non-dexamethasone-treated osteoblasts (92%) as compared to dexamethasone-treated osteoblasts after 1 (72%), 2 (63%), and 4 days (53%) of exposure. Dexamethasone-treated osteoblasts were viable and capable of proliferation, suggesting that the reduction of cellular attachment may be related to these cell adhesion processes. Immunofluorescent microscopy of both dexamethasone-treated osteoblasts and non-dexamethasone-treated osteoblasts failed to show any relative difference in the disassembly of focal adhesions and actin filaments. Extended dexamethasone treatment periods (up to 3 weeks) showed changes in the levels of FAK and FAK-phosphotyrosine in human embryonic palatal mesenchymal-1486 osteoblasts. The protocol used in this study demonstrated a glucocorticoid-induced osteoporosis-like suppression of osteoblasts. FAK disassembly was not a significant factor in short period; however, FAK protein levels and phosphotyrosine signaling on FAK were affected after 1-week exposure to dexamethasone. Phosphorylated FAK was not associated with the rise in the level of FAK, further indicating the possibility of FAK involvement in reduced cell attachment.


Neoplastic pleural effusion and intrathoracic metastasis of a scapular osteosarcoma in a dog: a multidisciplinary integrated diagnostic approach.
A 10-year-old, female spayed mixed-breed or cross-bred dog was referred to the Small Animal Teaching Hospital of the University of Liverpool due to tachypnea, dyspnea, and pleural effusion not responding to diuretics and antibiotics. The chest was drained and cytology of the pleural fluid was consistent with a modified transudate with presence of atypical cells initially attributed to mesothelial hyperplasia and dysplasia. Computed tomography detected, in addition to the bilateral pleural effusion, diffuse pleural thickening, multiple pleural and pulmonary nodules, and a mineralized and lytic mass in the left scapula. Imaging findings were suggestive of a primary bone tumor with intrathoracic metastasis. Cytology of the left scapular and pleural masses revealed a malignant neoplasm highly suggestive of osteosarcoma. The diagnosis was confirmed by demonstration of a positive cytochemical reaction for alkaline phosphatase on prestained cytology slides. This finding prompted review of the initial interpretation of the pleural effusion cytology. The presence of neoplastic osteoblasts in the thoracic fluid was identified by a combination of cytochemistry, cell pellet immunohistochemistry, and transmission electron microscopy findings. In this report, a multidisciplinary integrated diagnostic approach was used to diagnose and confirm a neoplastic pleural effusion due to osteosarcoma metastasis in a dog.


Development of a novel immuno-PCR assay for detection of ricin in ground beef, liquid chicken egg, and milk.
Reliable, sensitive, and high-throughput methods are essential for food defense, to detect foodborne contaminants and to facilitate remediation and recovery from potential toxin-related incidents. Ricin is a protein toxin that has been used for intentional contamination of foods in the past. In this study, we developed procedures for quantification of ricin in foods using immuno-PCR (IPCR). The direct adsorption of ricin onto the wells of a microtitration plate was compared with indirect immobilization via a capture antibody (sandwich IPCR). The latter procedure provided much greater sensitivity. We also compared a protocol with the immunoassay and PCR conducted in a single plate to a two-step procedure in which the PCR was conducted in a second plate, following release and transfer of the DNA marker. The two-step procedure proved 1,000-fold more sensitive for ricin detection, so this format was used to detect ricin in spiked samples of ground beef, chicken egg, and milk, and the results were compared with those obtained from enzyme-linked immunosorbent assay (ELISA). The IPCR had a limit of detection of 10 pg/ml in chicken egg and milk samples and 100 pg/ml in ground beef extracts. Comparable ELISA results were in the 1 to 10 ng/ml range. Thus, IPCR affords sensitivity that is 10-fold greater in the ground beef matrix, 100-fold greater in the milk, and 1,000-fold greater in the egg matrix than the sensitivity obtained by ELISA. Further optimization of the sandwich IPCR was performed by comparing various antibody combinations. Among the four formats investigated, the pAb-pAb combination yielded the lowest limit of detection (10 fg/ml).


Accumbens shell-hypothalamus interactions mediate extinction of alcohol seeking.
The nucleus accumbens shell (AcbSh) is required to inhibit drug seeking after extinction training. Conversely, the lateral hypothalamus (LH), which receives projections from AcbSh, mediates reinstatement of previously extinguished drug seeking. We hypothesized that reversible inactivation of AcbSh using GABA agonists (baclofen/muscimol) would reinstate extinguished alcohol seeking and increase neuronal activation in LH. Rats underwent self-administration training for 4% (v/v) alcoholic beer followed by extinction. AcbSh inactivation reinstated extinguished alcohol seeking when infusions were made after, but not before, extinction training. We then used immunohistochemical detection of c-Fos as a marker of neuronal activity, combined with immunohistochemical detection of the orexin and cocaine- and amphetamine-related transcript (CART) peptides, to study the profile and phenotype of neural activation during reinstatement produced by AcbSh inactivation. AcbSh inactivation increased c-Fos expression in hypothalamus, as well as in paraventricular thalamus and amygdala. Within hypothalamus, there was an increase in the number of orexin and CART cells expressing c-Fos. Finally, we hypothesized that concurrent inactivation of LH would prevent reinstatement produced by inactivation of AcbSh alone. Our results confirmed this. Together, these findings suggest that AcbSh mediates extinction of reward seeking by inhibiting hypothalamic neuropeptide neurons. Reversible inactivation of the AcbSh removes this influence, thereby releasing hypothalamus from AcbSh inhibition and enabling reinstatement of reward seeking. These ventral striatal-hypothalamic circuits for extinction overlap with those that mediate satiety, and we suggest that extinction training inhibits drug seeking because it co-opts neural circuits originally selected to produce satiety.


Transcriptome analysis of Houttuynia cordata Thunb. by Illumina paired-end RNA sequencing and SSR marker discovery.
Houttuynia cordata Thunb. is an important traditional medical herb in China and other Asian countries, with high medicinal and economic value. However, a lack of available genomic information has become a limitation for research on this species. Thus, we carried out high-throughput transcriptomic sequencing of H. cordata to generate an enormous transcriptome sequence dataset for gene discovery and molecular marker development. Illumina paired-end sequencing technology produced over 56 million sequencing reads from H. cordata mRNA. Subsequent de novo assembly yielded 63,954 unigenes, 39,982 (62.52%) and 26,122 (40.84%) of which had significant similarity to proteins in the NCBI nonredundant protein and Swiss-Prot databases (E-value <10(-5)), respectively. Of these annotated unigenes, 30,131 and 15,363 unigenes were assigned to gene ontology categories and clusters of orthologous groups, respectively. In addition, 24,434 (38.21%) unigenes were mapped onto 128 pathways using the KEGG pathway database and 17,964 (44.93%) unigenes showed homology to Vitis vinifera (Vitaceae) genes in BLASTx analysis. Furthermore, 4,800 cDNA SSRs were identified as potential molecular markers. Fifty primer pairs were randomly selected to detect polymorphism among 30 samples of H. cordata; 43 (86%) produced fragments of expected size, suggesting that the unigenes were suitable for specific primer design and of high quality, and the SSR marker could be widely used in marker-assisted selection and molecular breeding of H. cordata in the future. This is the first application of Illumina paired-end sequencing technology to investigate the whole transcriptome of H. cordata and to assemble RNA-seq reads without a reference genome. These data should help researchers investigating the evolution and biological processes of this species. The SSR markers developed can be used for construction of high-resolution genetic linkage maps and for gene-based association analyses in H. cordata. This work will enable future functional genomic research and research into the distinctive active constituents of this genus.


Single-substrate liquid-crystal displays by photo-enforced stratification.
Data visualization plays a crucial role in our society, as illustrated by the many displays that surround us. In the future, displays may become even more pervasive, ranging from individually addressable image-rendering wall hangings to data displays integrated in clothes. Liquid-crystal displays (LCDs) provide most of the flat-panel displays currently used. To keep pace with the ever-increasing possibilities afforded by developments in information technology, we need to develop manufacturing processes that will make LCDs cheaper and larger, with more freedom in design. Existing batch processes for making and filling LCD cells are relatively expensive, with size and shape limitations. Here we report a cost-effective, single-substrate technique in which a coated film is transformed into a polymer-covered liquid-crystal layer. This approach is based on photo-enforced stratification: a two-step photopolymerization-induced phase separation of a liquid-crystal blend and a polymer precursor. The process leads to the formation of micrometre-sized containers filled with a switchable liquid-crystal phase. In this way, displays can be produced on a variety of substrates using current coating technology. The developed process may be an important step towards new technologies such as 'display-on-anything' and 'paintable displays'.