Trends in coronary heart disease mortality and morbidity and related factors in Finland.
A marked increase in the coronary heart disease (CHD) mortality of working-age men and women occurred in Finland from the 1950s until the 1960s. Around the year 1970, CHD mortality started to decline and this decline still continues. In the age group 35-64 years the average annual decline of CHD mortality in the 1970s was 1.8% for men and 3.4% for women. Limited data available on trends in CHD morbidity show that the decline in CHD mortality is accompanied by a decline in the incidence of non-fatal myocardial infarction. CHD mortality and incidence are higher in east Finland than in west Finland and this east-west difference has so far persisted during the declining trend. The decline in CHD mortality and incidence in the 1970s has been preceded and paralleled by changes into favourable direction in dietary fat consumption and population mean levels for serum cholesterol, prevalence of smoking among adult Finnish men, control of hypertension by antihypertensive drug therapy, and management of patients with symptomatic CHD. Both the changes in life-styles and CHD risk factor levels, as well as changes in the management of patients with CHD, appear to have been contributing to the decline in CHD mortality and incidence in Finland.


Cutaneous mastocytosis combined with eruptive melanocytic nevi and melanoma. Coincidence or a linkage in the pathogenesis?
Cellular combinations in the same neoplasm can have intriguing physiopathological implications, which may be useful to better understand the biology of the diseases. Urticaria pigmentosa in association with eruptive melanocytic nevi was observed in a female patient. Maculopapular lesions extended at the base of different melanocytic nevi and the histopathological examination revealed the presence of a mast cell population in the papillary and reticular dermis combined with overlying melanocytic nevi. The re-evaluation of a melanoma removed three years before revealed the presence of the same pathological features. Immunohistochemical assays showed a strong positivity to Giemsa, Toluidine blue and CD-117 in the mast cells, while a S-100 reaction was observed in the melanocytic population. We discuss possible pathogenetic linkage between cutaneous mastocytosis and melanoma.


The safety and tolerability of newer antiepileptic drugs in children and adolescents.
Childhood epilepsy continues to be intractable in more than 25% of patients diagnosed with epilepsy. The introduction of new anti-epileptic drugs (AEDs) provides more options for treatment of children with epilepsy. We review the safety and tolerability of seven new AEDs (levetiracetam, lamotrigine, oxcarbazepine, rufinamide, topiramate, vigabatrin and zonisamide) focusing on their side effect profiles and safety in children and adolescents. Many considerations that are specific for children such as the impact of AEDs on the developing brain are not addressed during the development of new AEDs. They are usually approved as adjunctive therapies based upon clinical trials involving adult patients with partial epilepsy. However, 2 of the AEDs reviewed here (rufinamide and vigabatrin) have FDA approval in the U.S. for specific Pediatric epilepsy syndromes, which are discussed below. The Pediatrician or Neurologists decision on the use of a new AED is an evolutionary process largely dependent on the patient characteristics, personal/peer experiences and literature about efficacy and safety profiles of these medications. Evidence based guidelines are limited due to a lack of randomized controlled trials involving pediatric patients for many of these new AEDs.


Co-induction of jun B and c-fos in a subset of neurons in the spinal cord.
Noxious stimulation in vivo provokes the transcriptional activation of several genes which are thought to play an important role in the phenomena of stress and pain. In the rat, the expression of the c-fos proto-oncogene is rapidly induced upon noxious stimulation in defined neurons in the dorsal horn of the spinal cord. Interestingly, expression of the prodynorphin gene, which is thought to be involved in the endogenous mechanisms for pain/stress control, also localizes in the same anatomical area. Fos proteins are known to associate in transcriptional complexes with the products of the jun family constituting nuclear factor AP-1. These considerations prompted us to analyse the expression of the jun gene family members c-jun, jun B and jun D in rats subjected to noxious stimulation. We present data indicating that in unstimulated animals the transcripts of the three genes are differentially expressed and abundant within the various laminas of the lumbar spinal cord. Surprisingly, upon stimulation only the jun B transcript is augmented, being co-localized with Fos in a subset of neurons of the medial dorsal horn.


Transcatheter aortic valve implantation in aortic coarctation.
A 77-year-old male patient was scheduled for transcatheter aortic valve implantation for symptomatic and severe aortic valve stenosis. Severe multidirectional kinking of the aorta based on aortic coarctation did not allow for the transfemoral, but only for the transapical approach. The procedure was complicated because of the technically challenging retrograde passage of the transfemorally inserted pig-tail catheter required for intraoperative angiography of the aortic root. Correct positioning of the pig-tail catheter into the ascending aorta was accomplished by use of a loop snare, which was advanced into the descending aorta via the antegrade route, passing the cardiac apex, the stenotic aortic valve, and the coarctation-associated kinking. The pig-tail catheter tip was manipulated into the loop snare, pulled traverse the coarctation, and released within the proximal ascending aorta. Subsequent procedures were uneventful and followed the standardized protocol. A 29 mm Edwards Lifescience transcatheter Sapien bioprosthesis was successfully implanted.


A limited form of Wegener's granulomatosis with bronchiolitis obliterans organizing pneumonitis-like variant in an ulcerative colitis patient.
A case of a 19-year-old man with ulcerative colitis (UC) developed multiple pulmonary nodular shadows with cavity formation, elevated perinuclear antinuclear cytoplasmic antibody (pANCA) and positive lymphocyte stimulation test by drug (DLST) for mesalazine suggesting mesalazine induced Wegener's granulomatosis (WG). Transbronchial biopsy specimens were consistent with WG and thoracoscopic biopsy specimens were consistent with bronchiolitis obliterans organizing pneumonitis (BOOP). We diagnosed WG with BOOP-like variant, which was induced by mesalazine. To our knowledge, this is the first report of a limited form of WG associated with orally administered mesalazine.


Platelet serotonin measures in migraine.
Serotonergic mechanisms play an important role in the pathogenesis of headache. To search for potential indicators of altered serotonin homeostasis in migraine, we have investigated three parameters of the platelet serotonin (5HT) system, platelet serotonin level (PSL), platelet serotonin uptake (PSU), and monoamine oxidase (MAO-B) activity, in a group of 55 patients with migraine and in 81 healthy controls. After platelet separation, PSL was determined fluorimetrically; PSU was measured by incubating aliquots of platelet-rich plasma with six concentrations of 14C-5-HT for 60 seconds at 37 degrees C, followed by vacuum filtration; platelet MAO-B activity (toward kynuramine as a substrate) was determined fluorimetrically. Values of the investigated measures, in patients versus controls, amounted to (mean +/- SD) 608 +/- 166 vs. 591 +/- 184 ng/10(9) platelets for PSL, 139 +/- 25 vs. 142 +/- 25 pmol 5HT/10(8) platelets/minute for Vmax of PSU, 376 +/- 62 vs. 404 +/- 72 nM for Km of PSU, and 15.8 +/- 5.1 vs. 14.3 +/- 5.7 nmol product/10(8) platelets/60 minutes for velocity of MAO-B. Mentioned parameters did not show statistical differences between patients and controls, with exception of a small difference in Km of PSU, reaching significance (P<0.01). After subgrouping of patients according to diagnosis (migraine with aura, migraine without aura, and migraine attack) and gender, no differences retained significance. Our results indicate the absence of a measurable disturbance in 5HT homeostasis in migraine, as shown by platelet 5HT parameters, and they question the suitability of the use of mentioned blood elements in this regard.


Diet, cognition, and Alzheimer's disease: food for thought.
The prevention of Alzheimer's disease (AD) has become a real challenge due to its rising prevalence and the lack of an effective cure. Diet and nutrients have gained significant interest as potentially modifiable protective factors. The aim of this review is to provide an updated summary of evidence related to the effect of diet and nutritional factors on the risk of AD and cognitive aging, and discuss the potential mechanisms and confounding factors involved. A search was conducted in Medline and Web of Knowledge for epidemiological and clinical studies in the international literature from January 2000 to February 2013 using combinations of the following keywords: "Alzheimer's disease", "mild cognitive impairment", "cognitive function", "dietary factors", "omega-3", "antioxidants", "B vitamins", "dietary patterns", and "Mediterranean diet". Data from observational studies point to a protective role for certain nutrients, such as omega-3 fatty acids, antioxidants or B vitamins, and dietary patterns (Mediterranean diet). However, data from randomized controlled trials do not show a consistent effect. Whether confounding factors such as age, disease stage, other dietary components, cooking processes, and other methodological issues explain the divergent results remains to be established. Moreover, if certain nutrients protect against dementia, it is as yet unknown whether they may have a general effect on brain vascular health or directly interfere with the etiopathogenesis of AD.


Development of a modified Cohen method of standard setting.
A new 'Cohen' approach to standard setting was recently described where the pass mark is calculated as 60% of the score of the student at the 95th percentile, after correcting for guessing. This article considers how two potential criticisms of the Cohen method can be addressed and proposes a modified version, with the assumptions tested using local data. The modified version removes the correction for guessing and uses the score of the 90th, rather than the 95th percentile student as the reference point, based on the cumulative density functions for 32 modules from one medical school; and incorporates an indirect criterion-referenced passing standard by changing the 60% multiplier to the ratio of the cut score to the score of the student at the 90th percentile on exams that have been standard set using modified Angoff. The assumption that the performance of the 90th percentile student is consistent over time holds for multiple choice questions. Applying the modified Cohen method to the 32 modules generally reduced the variation in failure rate across modules, compared to a fixed pass mark of 50%. The results suggest that the modified Cohen method holds much promise as an economical approach to standard setting.


Oxygen Venous Embolism After Hydrogen Peroxide Use During Percutaneous Nephrolithotomy.
Background: Hydrogen peroxide (H2O2) is a common antiseptic that is available without a prescription in the United States, and it is indicated for minor dermal abrasion; mouth, gum, or dental irritations; and removal of oral secretion. Several other medical uses have also been described, including clot dissolution during endoscopic gastrointestinal evaluation, cleansing of orthopedic surgical sites, and bladder irrigation. However, these uses of H2O2, as well as high-dose ingestion, have been associated with a wide variety of medical complications, including but not limited to air pulmonary embolism and stroke. Case Presentation: Our patient is a 51-year-old female with a medical history of hypertension, familial, hypercholesterolemia, gallstones, depression, coronary artery disease (identified on calcium study because of familial hypercholesterolemia), nephrolithiasis, and recurrent cystitis. She required percutaneous nephrolithotomy and had H2O2 administered for clot dissolution. The clinical and temporal evidence would suggest a transient pulmonary air embolus after the intrarenal administration of or irrigation with H2O2, large amounts under high pressure. Conclusion: This represents the first reported incidence of air embolus as a result of intrarenal administration of H2O2.


Incidental ureteral fibroepithelial polyp.
Although rare, fibroepithelial polyp of the ureter is the most common type of benign urinary tract mesodermal tumor. It may cause symptoms such as hematuria, dysuria, and flank pain or be asymptomatic. Here we report incidental intraoperative detection of a left ureteral fibroepithelial polyp during an elective gyno-oncological surgery for left-sided ovarian thecoma.


The influence of glucose-lowering therapies on cancer risk in type 2 diabetes.
The risk of developing a range of solid tumours is increased in type 2 diabetes, and may be influenced by glucose-lowering therapies. We examined the risk of development of solid tumours in relation to treatment with oral agents, human insulin and insulin analogues. This was a retrospective cohort study of people treated in UK general practices. Those included in the analysis developed diabetes >40 years of age, and started treatment with oral agents or insulin after 2000. A total of 62,809 patients were divided into four groups according to whether they received monotherapy with metformin or sulfonylurea, combined therapy (metformin plus sulfonylurea), or insulin. Insulin users were grouped according to treatment with insulin glargine, long-acting human insulin, biphasic analogue and human biphasic insulin. The outcome measures were progression to any solid tumour, or cancer of the breast, colon, pancreas or prostate. Confounding factors were accounted for using Cox proportional hazards models. Metformin monotherapy carried the lowest risk of cancer. In comparison, the adjusted HR was 1.08 (95% CI 0.96-1.21) for metformin plus sulfonylurea, 1.36 (95% CI 1.19-1.54) for sulfonylurea monotherapy, and 1.42 (95% CI 1.27-1.60) for insulin-based regimens. Adding metformin to insulin reduced progression to cancer (HR 0.54, 95% CI 0.43-0.66). The risk for those on basal human insulin alone vs insulin glargine alone was 1.24 (95% CI 0.90-1.70). Compared with metformin, insulin therapy increased the risk of colorectal (HR 1.69, 95% CI 1.23-2.33) or pancreatic cancer (HR 4.63, 95% CI 2.64-8.10), but did not influence the risk of breast or prostate cancer. Sulfonylureas were associated with a similar pattern of risk as insulin. Those on insulin or insulin secretagogues were more likely to develop solid cancers than those on metformin, and combination with metformin abolished most of this excess risk. Metformin use was associated with lower risk of cancer of the colon or pancreas, but did not affect the risk of breast or prostate cancer. Use of insulin analogues was not associated with increased cancer risk as compared with human insulin.


Effects of reduced levels of sulfite in wine production using mixtures with lysozyme and dimethyl dicarbonate on levels of volatile and biogenic amines.
Sulphur dioxide (SO2) is an important preservative for wine, but its presence in foods can cause allergies and this has given impetus to the research for alternatives. The aim of this study was to reduce levels of sulfite in wine production using mixtures with lysozyme and dimethyl dicarbonate and examine the influence on levels of volatile and biogenic amines. To do so, vinifications were carried out using lysozyme, dimethyl dicarbonate (DMDC) and mixtures of these with SO2 in different concentrations (25 and 50 mg l-1). Results were compared with a control vinification with only SO2 (50 mg l-1). Mixing low concentrations of SO2 with lysozyme and DMDC reduced the concentration of biogenic amines (histamine, tyramine, putrescine, cadaverine, phenylethylamine + spermidine and spermine). In general, the total concentration of volatile amines (dimethylamine, isopropylamine, isobutylamine, pyrrolidine, ethylamine, diethylamine, amylamine and hexylamine) was higher in the sample fermented only with SO2. The concentrations of amines with secondary amino groups (dimethylamine, diethylamine, pyrrolidine) were higher in the sample only fermented with SO2 than those fermented with DMDC and lysozyme or with a mixture of preservatives. When SO2 was the only preservative in wine, total amine concentration (biogenic and volatile amines) was higher than for the rest of the treatments. Lysozyme by itself, and lysozyme mixed with SO2, both reduced the formation of biogenic amines but given the antioxidant activity of SO2 the use of the preservative mixture seems more advisable.


Experiences with carrier-mediated transport in liquid-phase microextraction.
Different organic borates, phosphates, sulphates, and carboxylic acids are evaluated as extraction carriers in three-phase liquid-phase microextraction (LPME). Hydrophilic basic drugs form ion-pairs with the carriers and are extracted as ion-pair complexes into an organic liquid membrane of n-octanol or peppermint oil immobilized in the pores of a polypropylene hollow fiber. From this point, the basic drugs are released into a 20-microL solution of 50mM HCl placed inside the lumen of the hollow fiber (acceptor solution). Simultaneously, the carrier is neutralized by protons from the acceptor solution (protonated to maintain the charge balance). Both water-soluble and water-insoluble carriers are tested. One promising candidate among the water-soluble carriers is 1-heptanesulfonic acid. This is added to the sample solution to a final concentration of 25mM and served to ion-pair the analytes within the sample solution. Among the less water-soluble candidates, a mixture of di(2-ethylhexyl) phosphate (DEHP) and tris(2-ethylhexyl) phosphate (TEHP) serve as efficient carriers. Ten percent (w/w) of each of DEHP and TEHP are added to the organic liquid membrane, and these carriers principally worked through ion-pairing with the analytes at the interface between the sample solution and the organic liquid membrane. Several carriers are found to be compatible with human plasma samples, and bromthymol blue is particularly efficient in combination with these protein-containing matrices. Following optimization of the conditions for bromthymol blue, including saturation of the plasma samples with sodium sulphate, extraction recoveries between 45% and 75% are obtained for eight model drugs after 60 min of extraction. With bromthymol blue as the carrier, highly acceptable validation data are obtained for phenylpropanolamine and practolol extracted from human plasma.


Locally advanced rectal adenocarcinoma: Are preoperative short and long course radiotherapy truly equivalent?
In the neoadjuvant treatment of locally advanced rectal adenocarcinoma, long- and short-course radiotherapy are considered to be of equivalent efficacy based upon several randomized trials. The present study assessed the effect of radiotherapy dose on overall survival. Using the National Cancer Database (2006-2013) 458 individuals with clinical stage II/III rectal adenocarcinoma treated were identified, with either short- (25 Gy) or long- (45 or 50.4 Gy) course neoadjuvant radiotherapy followed by surgery, without neoadjuvant or adjuvant chemotherapy. Multivariate COX regression was employed to evaluate differences in overall survival according to radiotherapy regimen. An association with improved overall survival in individuals treated with long- compared with short-course radiotherapy was demonstrated (HR=0.50, 0.34-0.73). The 30- and 90-day post-surgery mortality rates were higher in the short-course group when compared with the long-course group (12.2 vs. 2.4%; and 18.5 vs. 5.4%, respectively). Following the exclusion of patients that succumbed within 90-days post-surgery, overall survival advantage in the long-course group compared with the short-course group was maintained [hazard ratio (HR)=0.62, 0.39-0.99], with a median overall survival of 25.3 months (IQR 16.9-41.6) for the short-course group compared with 43.5 months (IQR 25.6-67.9) for the long-course group. To the best of our knowledge, the present results suggest for the first time that long-course radiotherapy is associated with an improved overall survival compared with short-course radiotherapy in locally advanced rectal adenocarcinoma in the absence of chemotherapy usage. This possible advantage is clinically relevant mainly in patients who cannot tolerate systemic chemotherapy.


Investigation into the molecular and thermodynamic basis of protein interactions in multimodal chromatography using functionalized nanoparticles.
Although multimodal chromatography offers significant potential for bioseparations, there is a lack of molecular level understanding of the nature of protein binding in these systems. In this study a nanoparticle system is employed that can simulate a chromatographic resin surface while also being amenable to isothermal titration calorimetry (ITC) and solution NMR. ITC and NMR titration experiments are carried out with (15)N-labeled ubiquitin to investigate the interactions of ubiquitin with nanoparticles functionalized with two industrially important multimodal ligands. The ITC results suggest that binding to both multimodal ligand surfaces is entropically driven over a range of temperatures and that this is due primarily to the release of surface bound waters. In order to reveal structural details of the interaction process, binding-induced chemical shift changes obtained from the NMR experiments are employed to obtain dissociation constants of individual amino acid residues on the protein surface. The residue level information obtained from NMR is then used to identify a preferred binding face on ubiquitin for interaction to both multimodal ligand surfaces. In addition, electrostatic potential and spatial aggregation propensity maps are used to determine important protein surface property data that are shown to correlate well with the molecular level information obtained from NMR. Importantly, the data demonstrate that the cluster of interacting residues on the protein surface act co-operatively to give rise to multimodal binding affinities several orders of magnitude greater than those obtained previously for interactions with free solution ligands. The use of NMR and ITC to study protein interactions with functionalized nanoparticles offers a new tool for obtaining important molecular and thermodynamic insights into protein affinity in multimodal chromatographic systems.


IRMPD spectroscopic study of microsolvated [Na(GlyAla)]+ and [Ca(GlyAla-H)]+ and the blue shifting of the hydrogen-bonded amide stretch with each water addition.
In this study, the structures of [Na(GlyAla)(H2O)](+) and [Ca(GlyAla-H)(H2O)n](+), (n = 1-3) solvated ion-molecule complexes (as well as the AlaGly isomers) were investigated using infrared multiple photon dissociation (IRMPD) spectroscopy and with computational methods. Calculations showed that in the calcium clusters, the lowest-energy complex is the one in which the peptide is deprotonated at the carboxylic acid end and that Ca(2+) binds to both carboxylate oxygen atoms as well as the amide carbonyl oxygen. For the microsolvated structures, all three water molecules also bind directly to Ca(2+). For the singly, doubly, and triply solvated complexes, these structures are supported by experimental IRMPD spectra. For the [Na(GlyAla)(H2O)](+) complex, both carbonyl oxygen atoms, one from the intact carboxylic acid and one from the amide group, as well as the water molecule were found to be bound to the Na(+). In all of the spectra, a strong band is observed between 3300 and 3400 cm(-1) and is assigned to the amide N-H stretch, which is red-shifted due to hydrogen bonding with the amine nitrogen. The position of the hydrogen-bonded amide N-H stretch is experimentally and theoretically found to be sensitive to the number of water molecules; it is shown to blue shift upon successive hydrations.


Identifying cow - level factors and farm characteristics associated with locomotion scores in dairy cows using cumulative link mixed models.
Lameness is a tremendous problem in intensively managed dairy herds all over the world. It has been associated with considerable adverse effects on animal welfare and economic viability. The majority of studies have evaluated factors associated with gait disturbance by categorising cows into lame and non-lame. This procedure yet entails a loss of information and precision. In the present study, we extend the binomial response to five categories acknowledging the ordered categorical nature of locomotion assessments, which conserves a higher level of information. A cumulative link mixed modelling approach was used to identify factors associated with increasing locomotion scores. The analysis revealed that a low body condition, elevated somatic cell count, more severe hock lesions, increasing parity, absence of pasture access, and poor udder cleanliness were relevant variables associated with higher locomotion scores. Furthermore, distinct differences in the locomotion scores assigned were identified in regard to breed, observer, and season. Using locomotion scores rather than a dichotomised response variable uncovers more refined relationships between gait disturbances and associated factors. This will help to understand the intricate nature of gait disturbances in dairy cows more deeply.


Angiotensin II, hypertension and angiotensin II receptor antagonism: Roles in the behavioural and brain pathology of a mouse model of Alzheimer's disease.
Elevated angiotensin II causes hypertension and contributes to Alzheimer's disease by affecting cerebral blood flow. Angiotensin II receptor blockers may provide candidates to reduce (vascular) risk factors for Alzheimer's disease. We studied effects of two months of angiotensin II-induced hypertension on systolic blood pressure, and treatment with the angiotensin II receptor blockers, eprosartan mesylate, after one month of induced hypertension in wild-type C57bl/6j and AβPPswe/PS1ΔE9 (AβPP/PS1/Alzheimer's disease) mice. AβPP/PS1 showed higher systolic blood pressure than wild-type. Subsequent eprosartan mesylate treatment restored this elevated systolic blood pressure in all mice. Functional connectivity was decreased in angiotensin II-infused Alzheimer's disease and wild-type mice, and only 12 months of Alzheimer's disease mice showed impaired cerebral blood flow. Only angiotensin II-infused Alzheimer's disease mice exhibited decreased spatial learning in the Morris water maze. Altogether, angiotensin II-induced hypertension not only exacerbated Alzheimer's disease-like pathological changes such as impairment of cerebral blood flow, functional connectivity, and cognition only in Alzheimer's disease model mice, but it also induced decreased functional connectivity in wild-type mice. However, we could not detect hypertension-induced overexpression of Aβ nor increased neuroinflammation. Our findings suggest a link between midlife hypertension, decreased cerebral hemodynamics and connectivity in an Alzheimer's disease mouse model. Eprosartan mesylate treatment restored and beneficially affected cerebral blood flow and connectivity. This model could be used to investigate prevention/treatment strategies in early Alzheimer's disease.


Endothelin-1 induces NAD(P)H oxidase in human endothelial cells.
Superoxide anions (O(*-)(2)) induce oxidative stress and reduce endothelial NO availability by peroxynitrite formation. In human endothelial cells gp91(phox) was identified as the limiting subunit of the forming NAD(P)H oxidase. Because endothelin-1 (ET-1) is considered as a pro-atherosclerotic stimulus, we analyzed the effect of ET-1 on gp91(phox) expression and O(*-)(2) generation in primary cultures of human umbilical vein endothelial cells (HUVECs). The gp91(phox) mRNA expression was quantified by standard calibrated competitive reverse transcriptase-polymerase chain reaction. ET-1 induces gp91(phox) mRNA expression in HUVEC (max. after 1 h). The induction of gp91(phox) expression was dose-dependent, reaching its maximum at 10 nmol/L ET-1. The increased gp91(phox) expression is mediated by endothelial receptor type B (ET(B)). Furthermore, ET-1 augments O(*-)(2) generation in human endothelial cells as measured by coelenterazine chemiluminescence. These data support a new mechanism: how ET-1 increases oxidative stress in the vessel wall leading to endothelial dysfunction and enhanced susceptibility to atherosclerosis.


A fingerprint marker from early gestation associated with diabetes in middle age: the Dutch Hunger Winter Families Study.
Fetal programming of diabetes might originate in early pregnancy when fingerprints are permanently established. The mean dermatoglyphic ridge count difference between fingertips 1 and 5 ('Md15') varies with the early prenatal environment. We hypothesized that Md15 would be associated with adult-onset diabetes. We obtained Md15 from 577 Dutch adults (aged 58.9 years, SD 1.1) whose births in 1943-47 were documented in maternity records and from 260 of their same-sex siblings for whom birth weights were not available. Of these 837 participants, complete anthropometry and diabetes status (from history or glucose tolerance test) were obtained for 819. After adjustment for age, sex, parental diabetes and adult anthropometry, fingerprint Md15 was associated with prevalent diabetes [odds ratio (OR) = 1.37 per 1 SD (95% confidence interval 1.02-1.84)]. This relationship held [OR = 1.40 (1.03-1.92)] for diabetic cases restricted to those recently diagnosed (within 7 years). In the birth series restricted to recently diagnosed cases, the mutually adjusted ORs were 1.34 (1.00-1.79) per SD of Md15 and 0.83 (0.62-1.10) per SD of birth weight. Further adjustments for maternal smoking, conception season or prenatal famine exposure in 1944-45 did not alter these estimates. Among 42 sibling pairs discordant for diabetes, the diabetic sibling had higher Md15 by 3.5 (0.6-6.3) after multivariable adjustment. Diabetes diagnosed at age 50+ years was associated with a fingerprint marker established in early gestation, irrespective of birth weight. Fingerprints may provide a useful tool to investigate prenatal developmental plasticity.


Ionic crystals [M3O(OOCC6H5)6(H2O)3]4[α-SiW12O40] (M = Cr, Fe) as heterogeneous catalysts for pinacol rearrangement.
Complexation of trinuclear oxo-centered carboxylates with a silicododecatungstate resulted in the formation of ionic crystals of [M(3)O(OOCC(6)H(5))(6)(H(2)O)(3)](4)[α-SiW(12)O(40)]·nH(2)O·mCH(3)COCH(3) [M = Cr (Ia), Fe (IIa)]. Treatments of Ia and IIa at 373 K in vacuo formed guest-free phases Ib and IIb, respectively. Compounds Ib and IIb heterogeneously catalyzed the pinacol rearrangement to pinacolone with high conversion at 373 K, and the catalysis is suggested to proceed size selectively in the solid bulk.


Spontaneous activity in peripheral diaphragmatic lymphatic loops.
The spontaneous contractility of FITC-dextran-filled lymphatics at the periphery of the pleural diaphragm was documented for the first time "in vivo" in anesthetized Wistar rats. We found that lymphatic segments could be divided into four phenotypes: 1) active, displaying rhythmic spontaneous contractions (51.8% of 197 analyzed sites); 2) stretch-activated, whose contraction was triggered by passive distension of the vessel lumen (4.1%); 3) passive, which displayed a completely passive distension (4.5%); and 4) inert, whose diameter never changed over time (39.6%). Smooth muscle actin was detected by immunofluorescence and confocal microscopy in the vessel walls of active but also of inert sites, albeit with a very different structure within the vessel wall. Indeed, while in active segments, actin was arranged in a dense mesh completely surrounding the lumen, in inert segments actin decorated the vessels wall in sparse longitudinal strips. When located nearby along the same lymphatic loop, active, stretch-activated, and passive sites were always recruited in temporal sequence starting from the active contraction. The time delay was ∼0.35 s between active and stretch-activated and 0.54 s between stretch-activated and passive segments, promoting a uniform lymph flux of ∼150/200 pl/min. We conclude that, unlike more central diaphragmatic lymphatic vessels, loops located at the extreme diaphragmatic periphery do require an intrinsic pumping mechanism to propel lymph centripetally, and that such an active lymph propulsion is attained by means of a complex interplay among sites whose properties differ but are indeed able to organize lymph flux in an ordered fashion.


Intraoperative radiation therapy in malignant glioma: early clinical results.
Intraoperative radiation therapy (IORT) with high energy electron beams is a treatment modality that has been included in multimodal programs in oncology to improve local tumor control. From August 1991 to December 1993, 17 patients with primary (8) or recurrent (9) high grade malignant gliomas, anaplastic astrocytoma (4), anaplastic oligodendroglioma (6) and glioblastoma multiforme (7), underwent surgical resection and a single dose of 10-20 Gy intraoperative radiation therapy was delivered in tumor bed. Fourteen patients received either pre-operative (8) or post-operative (6) external beam radiation therapy. Primary gliomas: 18-months actuarial survival rate has been 56% (range: 1-21+ months) and the median survival time has not yet been achieved. Four patients developed tumor progression (median time to tumor progression: 9 months). Recurrent gliomas: 18-months actuarial survival rate and median survival time has been 47% and 13 months (range: 6-32+ months) respectively. The median time to tumor progression was 11 months. No IORT related mortality has been observed. IORT is an attractive, tolerable and feasible treatment modality as antitumoral intensification procedure in high grade malignant gliomas.


AML1/RUNX1 works as a negative regulator of c-Mpl in hematopoietic stem cells.
In this study, we analyzed the roles for AML1/RUNX1 in the regulation of the c-mpl promoter. Wild-type AML1 activated the c-mpl promoter through the proximal AML-binding site in luciferase assays using 293T and HeLa cells. In accord with this result, electrophoretic mobility shift assay and chromatin immunoprecipitation assays demonstrated that AML1 bound to this site. Next, we analyzed the function of AML1 using a mutant of AML1 lacking the C terminus (AML1dC), which was originally found in a patient with myelodysplastic syndromes. AML1dC dominant-negatively suppressed transcriptional activity of wild-type AML1. However, unexpectedly, AML1dC-transduced murine c-Kit(+)Sca1(+)Lineage(-) cells expressed c-mpl mRNA and c-Mpl protein more abundantly than mock-transduced cells, which led to the enhanced thrombopoietin-mediated proliferation. Moreover, when AML1dC was induced to express during the development of hematopoietic cells from embryonic stem (ES) cells, AML1dC augmented the c-Mpl expression on hematopoietic stem/progenitor cells. Furthermore, we found that early hematopoietic cells that derived from AML1(+/-) ES cells expressed c-Mpl more intensely than those that developed from wild-type ES cells. In contrast, AML1dC hardly affected c-Mpl expression and maturation of megakaryocytes. As for the mechanism of the different roles of AML1 in the regulation of the c-mpl promoter, we found that AML1 forms a complex with a transcription repressor mSin3A on the c-mpl promoter in hematopoietic stem/progenitor cells, although it forms a complex with a transcription activator p300 on the same promoter in megakaryocytic cells. Together, these data indicate that AML1 can regulate the c-mpl promoter both positively and negatively by changing the binding partner according to cell types.


Molecular Characterization of Reduced Susceptibility to Biocides in Clinical Isolates of Acinetobacter baumannii.
Active efflux is regarded as a common mechanism for antibiotic and biocide resistance. However, the role of many drug efflux pumps in biocide resistance in Acinetobacter baumannii remains unknown. Using biocide-resistant A. baumannii clinical isolates, we investigated the incidence of 11 known/putative antimicrobial resistance efflux pump genes (adeB, adeG, adeJ, adeT1, adeT2, amvA, abeD, abeM, qacE, qacEΔ1, and aceI) and triclosan target gene fabI through PCR and DNA sequencing. Reverse transcriptase quantitative PCR was conducted to assess the correlation between the efflux pump gene expression and the reduced susceptibility to triclosan or chlorhexidine. The A. baumannii isolates displayed high levels of reduced susceptibility to triclosan, chlorhexidine, benzalkonium, hydrogen peroxide, and ethanol. Most tested isolates were resistant to multiple antibiotics. Efflux resistance genes were widely distributed and generally expressed in A. baumannii. Although no clear relation was established between efflux pump gene expression and antibiotic resistance or reduced biocide susceptibility, triclosan non-susceptible isolates displayed relatively increased expression of adeB and adeJ whereas chlorhexidine non-susceptible isolates had increased abeM and fabI gene expression. Increased expression of adeJ and abeM was also demonstrated in multiple antibiotic resistant isolates. Exposure of isolates to subinhibitory concentrations of triclosan or chlorhexidine induced gene expression of adeB, adeG, adeJ and fabI, and adeB, respectively. A point mutation in FabI, Gly95Ser, was observed in only one triclosan-resistant isolate. Multiple sequence types with the major clone complex, CC92, were identified in high level triclosan-resistant isolates. Overall, this study showed the high prevalence of antibiotic and biocide resistance as well as the complexity of intertwined resistance mechanisms in clinical isolates of A. baumannii, which highlights the importance of antimicrobial stewardship and resistance surveillance in clinics.


Exploring the process of progressive disorientation.
While an increasing number of behavioral studies examining spatial cognition use experimental paradigms involving disorientation, the process by which one becomes disoriented is not well explored. The current study examined this process using a paradigm in which participants were blindfolded and underwent a succession of 70 degrees or 200 degrees passive, whole body rotations around a fixed vertical axis. After each rotation, participants used a pointer to indicate either their heading at the start of the most recent turn or their heading at the start of the current series of turns. Analyses showed that in both cases, mean pointing errors increased gradually over successive turns. In addition to the gradual loss of orientation indicated by this increase, analysis of the pointing errors also showed evidence of occasional, abrupt loss orientation. Results indicate multiple routes from an oriented to a disoriented state, and shed light on the process of becoming disoriented.


Smartphone-Based Electrochemical Biosensors for Directly Detecting Serum-Derived Exosomes and Monitoring Their Secretion.
Exosomes are potential biomarkers, which play an important role in early diagnosis and prognosis prediction of cancer-related diseases. Nevertheless, direct quantification of exosomes in biological fluid, especially in point-of-care tests (POCTs), remains extremely challenging. Herein, we developed a sensitive and portable electrochemical biosensor in combination with smartphones for quantitative analysis of exosomes. The improved double-antibody sandwich method-based poly-enzyme signal amplification was adopted to detect exosomes. We could detect as low as 7.23 ng of CD63-positive exosomes in 5 μL of serum within 2 h. Importantly, we demonstrated that the biosensor worked well with microliter-level serum and cell culture supernatant. The biosensor holds great potential for the detection of CD-63-expressing exosomes in early diagnosis of prostate disease because CD63-positive exosomes were less detected from the prostate patient serum. Also, the biosensor was used to monitor the secretion of exosomes with the drug therapy, showing a close relationship between the secretion of exosomes and the concentration of cisplatin. The biosensing platform provides a novel way toward POCT for the diagnosis and prognosis prediction of prostate disease and other diseases via biomarker expression levels of exosomes.


Common molecular determinants of local anesthetic, antiarrhythmic, and anticonvulsant block of voltage-gated Na+ channels.
Voltage-gated Na+ channels are the molecular targets of local anesthetics, class I antiarrhythmic drugs, and some anticonvulsants. These chemically diverse drugs inhibit Na+ channels with complex voltage- and frequency-dependent properties that reflect preferential drug binding to open and inactivated channel states. The site-directed mutations F1764A and Y1771A in transmembrane segment IVS6 of type IIA Na+ channel alpha subunits dramatically reduce the affinity of inactivated channels for the local anesthetic etidocaine. In this study, we show that these mutations also greatly reduce the sensitivity of Na+ channels to state-dependent block by the class Ib antiarrhythmic drug lidocaine and the anticonvulsant phenytoin and, to a lesser extent, reduce the sensitivity to block by the class Ia and Ic antiarrhythmic drugs quinidine and flecainide. For lidocaine and phenytoin, which bind preferentially to inactivated Na+ channels, the mutation F1764A reduced the affinity for binding to the inactivated state 24.5-fold and 8.3-fold, respectively, while Y1771A had smaller effects. For quinidine and flecainide, which bind preferentially to the open Na+ channels, the mutations F1764A and Y1771A reduced the affinity for binding to the open state 2- to 3-fold. Thus, F1764 and Y1771 are common molecular determinants of state-dependent binding of diverse drugs including lidocaine, phenytoin, flecainide, and quinidine, suggesting that these drugs interact with a common receptor site. However, the different magnitude of the effects of these mutations on binding of the individual drugs indicates that they interact in an overlapping, but nonidentical, manner with a common receptor site. These results further define the contributions of F1764 and Y1771 to a complex drug receptor site in the pore of Na+ channels.


Financial inclusion and its influence on renewable energy consumption-environmental performance: the role of ICTs in China.
Financial inclusion means that individuals and businesses can easily avail financial goods and services at an affordable cost. It is widely recognized that financial inclusion can help preserve the environment by increasing the consumption of renewable energy sources. Hence, our basic aim is to investigate whether financial inclusion has any effect on renewable energy consumption and environmental quality in China. To get the estimates of the variables, we have preferred the ARDL model. The estimates of the model confirm that a rise in the number of ATMs and total insurance has a positive impact on renewable energy consumption in China in the long run. Conversely, an increase in the number of ATMs and total insurance negatively affects CO2 emissions in China. In general, we can say that financial inclusion increases renewable energy consumption and reduces CO2 emissions in China. Therefore, by using financial inclusion, policymakers should try to divert the resources towards environmentally friendly consumption and production activities.


Spatial considerations in the resolution of inflammation: Elucidating leukocyte interactions via an experimentally-calibrated agent-based model.
Many common medical conditions (such as cancer, arthritis, chronic obstructive pulmonary disease (COPD), and others) are associated with inflammation, and even more so when combined with the effects of ageing and multimorbidity. While the inflammatory response varies in different tissue types, under disease and in response to therapeutic interventions, it has common interactions that occur between immune cells and inflammatory mediators. Understanding these underlying inflammatory mechanisms is key in progressing treatments and therapies for numerous inflammatory conditions. It is now considered that constituent mechanisms of the inflammatory response can be actively manipulated in order to drive resolution of inflammatory damage; particularly, those mechanisms related to the pro-inflammatory role of neutrophils and the anti-inflammatory role of macrophages. In this article, we describe the assembly of a hybrid mathematical model in which the spatial spread of inflammatory mediators is described through partial differential equations, and immune cells (neutrophils and macrophages) are described individually via an agent-based modelling approach. We pay close attention to how immune cells chemotax toward pro-inflammatory mediators, presenting a model for cell chemotaxis that is calibrated against experimentally observed cell trajectories in healthy and COPD-affected scenarios. We illustrate how variations in key model parameters can drive the switch from resolution of inflammation to chronic outcomes, and show that aberrant neutrophil chemotaxis can move an otherwise healthy outcome to one of chronicity. Finally, we reflect on our results in the context of the on-going hunt for new therapeutic interventions.


Extracellular cell wall β(1,3)glucan is required to couple septation to actomyosin ring contraction.
Cytokinesis has been extensively studied in different models, but the role of the extracellular cell wall is less understood. Here we studied this process in fission yeast. The essential protein Bgs4 synthesizes the main cell wall β(1,3)glucan. We show that Bgs4-derived β(1,3)glucan is required for correct and stable actomyosin ring positioning in the cell middle, before the start of septum formation and anchorage to the cell wall. Consequently, β(1,3)glucan loss generated ring sliding, oblique positioned rings and septa, misdirected septum synthesis indicative of relaxed rings, and uncoupling between a fast ring and membrane ingression and slow septum synthesis, suggesting that cytokinesis can progress with defective septum pushing and/or ring pulling forces. Moreover, Bgs4-derived β(1,3)glucan is essential for secondary septum formation and correct primary septum completion. Therefore, our results show that extracellular β(1,3)glucan is required for cytokinesis to connect the cell wall with the plasma membrane and for contractile ring function, as proposed for the equivalent extracellular matrix in animal cells.


Penicillin-binding proteins (PBP) and Lmo0441 (a PBP-like protein) play a role in Beta-lactam sensitivity of Listeria monocytogenes.
While seven penicillin-binding proteins (PBPs) or PBP-like proteins have been identified either by radiolabelled penicillin binding studies or genomic analysis, only PBP3 has been considered of interest for Beta-lactams activity against Listeria monocytogenes. Herein we reveal that both PBP4 and Lmo0441 (a PBP-like protein) play a direct role in cephalosporin activity in L. monocytogenes while PBP4 additionally has a protective affect against both penicillin and carbapenem.


Factors That Predispose Undergraduates to Mental Issues: A Cumulative Literature Review for Future Research Perspectives.
Distress and mental health issues among college students is an emerging topic of study. The aim of this research work is to illustrate academic and social risk factors and how they prove to be predictors of anxiety and depressive disorders. The methodology used is a cumulative literature review structured over 10 systematic phases, and is replicable. Showing considerable potential for cumulative research, the relevance of this study reflects the concern of the academic community and international governments. The articles selected range from categorization of disorders in relation to mental health, to reporting the condition of rhinestones and difficulties of students in university contexts. In conclusion, the research focusses upon predisposing, concurrent or protective factors relating to the mental health of university students, so that institutions can act on concrete dynamics or propose targeted research on this topic.


Facial reanimation surgery utilizing the Mitek anchor system: A case report.
A flaccid hemi-face is frequently the most noticeable and cosmetically unacceptable consequence of facial nerve palsy, whether due to trauma, Bell's palsy or other etiologies. A variety of face-lift and reanimation techniques have been utilized in the past, but with time, these frequently require further surgery. We describe the use of Mitek (Norwood, MA) suture anchors for cheek resuspension in a patient with facial palsy. This system is composed of a drill guide, drill, inserter, and anchor. Although the titanium alloy anchors come in multiple sizes, the Mini GII Anchor is typically most appropriate for use in facial procedures. The actual size of the Mini GII Anchor is 1.8 mm in diameter and 5.4 mm in length. Two small arched prongs extend from the body of the anchor, and an eyelet at the superior surface is used for suture placement. When placed into a pre-drilled hole with the insertion tool, the prongs extend, effectively fixing the anchor in place. The drill guide protects adjacent soft tissues during the drilling process and allows drilling to a predetermined fixed depth. Sutures attached to the anchor may then be used for soft tissue fixation to bone.


Structures of wild-type and selected CMT1X mutant connexin 32 gap junction channels and hemichannels.
In myelinating Schwann cells, connection between myelin layers is mediated by gap junction channels (GJCs) formed by docked connexin 32 (Cx32) hemichannels (HCs). Mutations in Cx32 cause the X-linked Charcot-Marie-Tooth disease (CMT1X), a degenerative neuropathy without a cure. A molecular link between Cx32 dysfunction and CMT1X pathogenesis is still missing. Here, we describe the high-resolution cryo-electron cryo-myography (cryo-EM) structures of the Cx32 GJC and HC, along with two CMT1X-linked mutants, W3S and R22G. While the structures of wild-type and mutant GJCs are virtually identical, the HCs show a major difference: In the W3S and R22G mutant HCs, the amino-terminal gating helix partially occludes the pore, consistent with a diminished HC activity. Our results suggest that HC dysfunction may be involved in the pathogenesis of CMT1X.


Toward Systems Metabolic Engineering of Streptomycetes for Secondary Metabolites Production.
Streptomycetes are known for their inherent ability to produce pharmaceutically relevant secondary metabolites. Discovery of medically useful, yet novel compounds has become a great challenge due to frequent rediscovery of known compounds and a consequent decline in the number of relevant clinical trials in the last decades. A paradigm shift took place when the first whole genome sequences of streptomycetes became available, from which silent or "cryptic" biosynthetic gene clusters (BGCs) were discovered. Cryptic BGCs reveal a so far untapped potential of the microorganisms for the production of novel compounds, which has spurred new efforts in understanding the complex regulation between primary and secondary metabolism. This new trend has been accompanied with development of new computational resources (genome and compound mining tools), generation of various high-quality omics data, establishment of molecular tools, and other strain engineering strategies. They all come together to enable systems metabolic engineering of streptomycetes, allowing more systematic and efficient strain development. In this review, the authors present recent progresses within systems metabolic engineering of streptomycetes for uncovering their hidden potential to produce novel compounds and for the improved production of secondary metabolites.


Hereditary leiomyomatosis and renal cell cancer syndrome associated renal cell carcinoma.
Hereditary leiomyomatosis and renal cell carcinoma (HLRCC) is a recently described entity with unknown exact prevalence. The affected individuals are predisposed to have multiple leiomyomas and renal cancer due to germline mutation in fumarate hydratase gene on chromosome 1. The knowledge of this rare tumour is essential for early recognition and institution of appropriate therapy, since they have a grave prognosis. Herein, we present the first case from India of HLRCC in a 42 year old lady who presented with a renal mass and metastasis with consequent fulminant course of disease. We discuss the detailed histomorphologic features and iunique immunohistochemical signature of this unusual renal tumour with discussion of differential diagnosis.


Salmonella bacteraemia occurring concurrently with the first presentation of systemic lupus erythematosus.
There is increasing recognition of the association between Salmonella infections and systemic lupus erythematosus (SLE). However, these infections have usually occurred in patients with established SLE who were receiving immunosuppressive therapy. We describe two young women with previously undiagnosed SLE who developed Salmonella bacteraemia. Neither of these patients had received immunosuppressive drugs. From these and two previously reported patients, it may be concluded that SLE per se is likely to be associated with a defect in host defences to Salmonella bacteria.


[The features of a functional state of the coronary circulation in women-smokers with non st elevation acute coronary syndrome].
Acute coronary syndrome is one of the most adverse prognostic clinical forms of ischemic heart disease. In recent years, attention of researchers and cardiologists practical attract female patients due to a substantial increase in the prevalence including acute coronary syndrome, differences in the causes and features of its course. Women often diagnosed myocardial infarction without ST elevation and nonobstructive coronary heart disease associated with coronary spasm or stratification of coronary artery. The study of the pathophysiological mechanisms of myocardial infarction also showed the existence of certain gender differences. However, the available literature contains little data on the impact of smoking on morphological and functional condition of the coronary arteries in women with severe forms of coronary heart disease. to conduct contrastive analysis of coronary circulation state according to data of coronarography (CAG) in women with non ST elevation acute coronary syndrome. 61 women aged 35-77 were examined (middle age 57, 9±1,2) who were hospitalized because of non ST elevation acute coronary syndrome. Depending on smoking habit all the patients of general group were divided into two subgroups. First subgroup (n=29, middle age 54,3±1,5) consisted of smoking women who had been smoking for 5 years ≥5 cigarettes a day (an average number of smoked cigarettes per day was 9,8) Second subgroup (n=32, middle age 61,8±1,3) consisted of ill non-smoking women. We studied a type of coronary circulation? its nature, localization and evidence of coronary arteria damage and characteristics of microvascular and collateral blood flow. smoking women with non ST elevation acute coronary syndrome more often have bigger damages of coronary arteria: hemodynamically relevant and diffusive damages mostly located in proximal medial segments often with connected damage of distal segments especially of the left coronary arteria and its interventricular branch that causes a compensatory hypernormal development collateral blood flow and predominance of right blood supply type. Approximately 59,02% of women have sustainable violation of coronary microcirculation and myocardial perfusion, especially in cases of absence of coronary arteria hemodynamically relevant stenosis; much more often (by half) it is observed in smoking women (75,86%) that can be recognized on a coronarography as rather delayed blood flow and retention of contrast release. One-third of women occurs in the setting of almost unchanged coronary red vessels. durable smoking causes hard vessel damages with the occurrence of hemodynamic stenosis in proximal and medial segments of magistral coronary arteries, which are often connected with their distal (diffusive) changes and more frequent violation in microcirculation system.


Achievement of disease control with donor-derived EB virus-specific cytotoxic T cells after allogeneic peripheral blood stem cell transplantation for aggressive NK-cell leukemia.
Aggressive NK-cell leukemia (ANKL) is characterized by systemic infiltration of Epstein-Barr virus (EBV)-associated natural killer cells and poor prognosis. We report a case of ANKL in which EBV-specific cytotoxic T lymphocytes (CTLs) were induced. A 41-year-old male suffered from fever, pancytopenia, and hepatosplenomegaly. The number of abnormal large granular lymphocytes in the bone marrow was increased and the cells were positive for CD56 and EBV-encoded small nuclear RNAs. The patient was diagnosed with ANKL and achieved a complete response following intensive chemotherapy. He then underwent allogeneic peripheral blood stem cell transplantation from his sister. Conditioning therapy consisted of total body irradiation and cyclophosphamide. Graft-versus-host disease prophylaxis consisted of cyclosporine and methotrexate. On day 31, complete donor chimerism was achieved and no acute graft-versus-host disease developed. The ANKL relapsed on day 80, and cyclosporine was rapidly tapered and chemotherapy was started. During hematopoietic recovery, the number of atypical lymphocytes increased, but they were donor-derived EBV-specific CTLs. The patient achieved a partial response and EBV viral load decreased to normal range. Unfortunately, ANKL worsen again when the CTLs disappeared from his blood. This is the first case report of ANKL in which induced EBV-specific CTLs may have contributed to disease control.


Postangioplasty restenosis rate between segments of the major coronary arteries.
Conflicting data have been published regarding the rate of postangioplasty restenosis observed in diverse segments of the coronary tree. However, these studies may be criticized for their biased selection of patients, methods of analysis, and definitions of restenosis. In the present study, 1,353 patients underwent a successful coronary dilatation of greater than or equal to 1 site. In all, 1,234 patients (91%) had a follow-up angiogram after 6 months, or earlier when indicated by symptoms. All films were processed and analyzed at the thoraxcenter core laboratory with the coronary angiography analysis system (automated contour detection). Restenosis was considered present if the diameter stenosis at follow-up was greater than 50%. No differences in restenosis rates were observed between coronary segments using this categorical definition. A continuous approach was also used; absolute changes in minimal luminal diameter adjusted for vessel size were used in order to allow comparison between vessels of different sizes (relative loss). No significant differences were observed between the coronary segments with this continuous approach. These results suggest that restenosis is a ubiquitous phenomenon without any predilection for a particular site in the coronary tree.


Dynamic reversal of random X-Chromosome inactivation during iPSC reprogramming.
Induction and reversal of chromatin silencing is critical for successful development, tissue homeostasis, and the derivation of induced pluripotent stem cells (iPSCs). X-Chromosome inactivation (XCI) and reactivation (XCR) in female cells represent chromosome-wide transitions between active and inactive chromatin states. Although XCI has long been studied, providing important insights into gene regulation, the dynamics and mechanisms underlying the reversal of stable chromatin silencing of X-linked genes are much less understood. Here, we use allele-specific transcriptomics to study XCR during mouse iPSC reprogramming in order to elucidate the timing and mechanisms of chromosome-wide reversal of gene silencing. We show that XCR is hierarchical, with subsets of genes reactivating early, late, and very late during reprogramming. Early genes are activated before the onset of late pluripotency genes activation. Early genes are located genomically closer to genes that escape XCI, unlike genes reactivating late. Early genes also show increased pluripotency transcription factor (TF) binding. We also reveal that histone deacetylases (HDACs) restrict XCR in reprogramming intermediates and that the severe hypoacetylation state of the inactive X Chromosome (Xi) persists until late reprogramming stages. Altogether, these results reveal the timing of transcriptional activation of monoallelically repressed genes during iPSC reprogramming, and suggest that allelic activation involves the combined action of chromatin topology, pluripotency TFs, and chromatin regulators. These findings are important for our understanding of gene silencing, maintenance of cell identity, reprogramming, and disease.


Purification and Properties of Arginase from Soybean, Glycine max, Axes.
Arginase (EC 22.214.171.124) was purified to homogeneity from cytosol of soybean, Glycine max, axes by chromatographic separations on Sephadex G-200, DEAE-sephacel, hydroxyapatite, and arginine-affinity columns. The molecular weight of the enzyme estimated by pore gradient gel electrophoresis was 240,000, while sodium dodecyl sulfate polyacrylamide gel electrophoresis gave a single band at the molecular weight of 60,000. The optimal pH for activity was 9.5 and the K(m) value was 83 millimolar. The enzyme was stimulated by polyamines such as putrescine.


[Analysis of the small supernumerary marker chromosome in Turner syndrome with 45, X/46, X, + mar karyotype].
To identify the origin and study the morphology of small supernumerary marker chromosome (sSMC) in Turner syndrome with 45, X/46, X, + mar karyotype. Using the conventional chromosome G-banding technique, 10 cases of Turner syndrome with 45, X/46, X, + mar chromosome karyotype were obtained, dual-color fluorescence in situ hybridization was applied to study the origin and morphology of the sSMC. In the 10 cases of Turner syndrome with 45, X/46, X, + mar karyotype, the sSMC of 7 cases was derived from X chromosome [sSMC(X)], the sSMC of 2 cases was derived from Y chromosome [sSMC(Y)] and the remaining 1 case was derived from the autosome. There were 4 cases of ring(r) chromosomes and 3 of centric minutes (min) in the 7 sSMC (X) cases. In the 2 sSMC(Y), one case was dicentric (dic) and the other was centric minute (min). The sSMC originated from the autosome was a centric minute (min). The origin of sSMC of Turner syndrome with 45, X/46, X, + mar karyotype was almost all from sex chromosomes, and rarely from autosomes. sSMC can exist as isodicentric, ring, or centric minute. The molecular cytogenetic features of the sSMC can provide useful information for genetic counseling, prenatal diagnosis and treatment of the Turner syndrome patients with a 45, X/46, X, + mar karyotype.


p38 Mitogen-activated protein kinase-, calcium-calmodulin-dependent protein kinase-, and calcineurin-mediated signaling pathways transcriptionally regulate myogenin expression.
In this report, we identify myogenin as an important transcriptional target under the control of three intracellular signaling pathways, namely, the p38 mitogen-activated protein kinase- (MAPK), calcium-calmodulin-dependent protein kinase- (CaMK), and calcineurin-mediated pathways, during skeletal muscle differentiation. Three cis-elements (i.e., the E box, myocyte enhancer factor [MEF] 2, and MEF3 sites) in the proximal myogenin promoter in response to these three pathways are defined. MyoD, MEF2s, and Six proteins, the trans-activators bound to these cis-elements, are shown to be activated by these signaling pathways. Our data support a model in which all three signaling pathways act in parallel but nonredundantly to control myogenin expression. Inhibition of any one pathway will result in abolished or reduced myogenin expression and subsequent phenotypic differentiation. In addition, we demonstrate that CaMK and calcineurin fail to activate MEF2s in Rhabdomyosarcoma-derived RD cells. For CaMK, we show its activation in response to differentiation signals and its effect on the cytoplasmic translocation of histone deacetylases 5 are not compromised in RD cells, suggesting histone deacetylases 5 cytoplasmic translocation is necessary but not sufficient, and additional signal is required in conjunction with CaMK to activate MEF2 proteins.


Circular harmonic averaging of rotary-shadowed and negatively stained creatine kinase macromolecules.
The structure of mitochondrial creatine kinase is investigated by high-resolution shadowing at very low temperature and conventional negative staining. The electron microscopic images are analyzed with circular harmonic averaging, a method suited for the processing of single molecules. The rotational alignment and averaging is performed with the circular harmonic components, which allows data compression and several steps of noise reduction to be carried out within the averaging procedure. In addition, the symmetry can be deduced. For the mitochondrial creatine kinase, a fourfold symmetry is found that is compatible with the biochemical and biophysical characterization of the molecule.


Mu and delta, but not kappa, opioid agonists induce contractions of the canine small intestine in vivo.
Extraluminal strain gage transducers were sutured along the transverse axis of the duodenum in order to monitor circular muscle contractile activity in the pentobarbital anesthetized dog. Administration by intravenous bolus of a variety of mu- and delta-directed opioid ligands resulted in a dose-dependent increase in duodenal contractile activity. In contrast, all kappa-directed ligands were devoid of stimulatory activity. Naloxone reversed the effects of normorphine and [Met5]enkephalin but was 20 times more effective against normorphine than [Met5]enkephalin. Based on the inactivity of all kappa ligands examined and the differential potency of naloxone against [Met5]enkephalin and normorphine, we suggest that this model may be useful in the classification of opioid ligands as to their receptor selectivity in vivo. Further, these data indicate that the stimulation of duodenal contractile activity is not mediated by enteric kappa receptors.


Allergen-free probiotics.
Food sensitivities are constantly increasing in "westernized" countries and may pose serious health risks to sensitized individuals. Severe allergy episodes have also been reported after the intake of probiotic products containing milk protein residues, especially in children. The need for safe and effective probiotic strains and food supplements, which contain them, is now emerging clearly. The present work describes the way of achieving this aim by the avoidance of any kind of raw materials at risk, both in probiotic strain industrial manufacturing and finished product formulation. Allergen-free probiotics represent, without any doubt, an innovative and safe tool for human health.


BIM mediates oncogene inactivation-induced apoptosis in multiple transgenic mouse models of acute lymphoblastic leukemia.
Oncogene inactivation in both clinical targeted therapies and conditional transgenic mouse cancer models can induce significant tumor regression associated with the robust induction of apoptosis. Here we report that in MYC-, RAS-, and BCR-ABL-induced acute lymphoblastic leukemia (ALL), apoptosis upon oncogene inactivation is mediated by the same pro-apoptotic protein, BIM. The induction of BIMin the MYC- and RAS-driven leukemia is mediated by the downregulation of miR-17-92. Overexpression of miR-17-92 blocked the induction of apoptosis upon oncogene inactivation in the MYC and RAS-driven but not in the BCR-ABL-driven ALL leukemia. Hence, our results provide novel insight into the mechanism of apoptosis upon oncogene inactivation and suggest that induction of BIM-mediated apoptosis may be an important therapeutic approach for ALL.


Hypnotic amnesia and learning: a dissociation interpretation.
The effect of hypnotically induced amnesia on positive transfer learning was examined within the conceptual framework of dissociation theory. An Experimental group learned two paired-associates word lists that contained highly similar (between the two lists) stimulus words. After learning the first list, subjects were given suggestions for amnesia for that list. Control group 1 learned the same two lists without amnesia suggestions. Control group 2, with amnesia suggestions following the first list, learned two lists that contained dissimilar stimulus words. All subjects remained hypnotized throughout the session. Positive transfer between the two lists was demonstrated for the Experimental group and for Control group 1, but not for Control group 2. The results are viewed as consistent with predictions from dissociation theory.


The effect of bezafibrate on hyperlipidaemia in experimental nephrotic syndrome in rats.
The effects of bezafibrate on hyperlipidaemia in experimental nephrotic syndrome in rats has been investigated. The treated group received bezafibrate 10 mg kg-1 p.o. daily. No significant differences in total serum cholesterol occurred, but a significant reduction in serum triglyceride (P less than 0.005) and elevation in HDL cholesterol (P less than 0.005) occurred. These findings may have implications for therapeutic intervention in severe hyperlipidaemia of the nephrotic syndrome in man.


The NordiNet® International Outcome Study and NovoNet® ANSWER Program®: rationale, design, and methodology of two international pharmacoepidemiological registry-based studies monitoring long-term clinical and safety outcomes of growth hormone therapy (Norditropin®).
Randomized controlled trials have shown that growth hormone (GH) therapy has effects on growth, metabolism, and body composition. GH therapy is prescribed for children with growth failure and adults with GH deficiency. Carefully conducted observational study of GH treatment affords the opportunity to assess long-term treatment outcomes and the clinical factors and variables affecting those outcomes, in patients receiving GH therapy in routine clinical practice. The NordiNet® International Outcome Study (IOS) and the American Norditropin® Web Enabled Research (ANSWER Program®) are two complementary, non-interventional, observational studies that adhere to current guidelines for pharmacoepidemiological data. The studies include pediatric and adult patients receiving Norditropin®, as prescribed by their physicians. The studies gather long-term data on the safety and effectiveness of reallife treatment with the recombinant human GH, Norditropin®. We describe the origins, aims, objectives, and design methodology of the studies, as well as their governance and validity, strengths, and limitations. The NordiNet® IOS and ANSWER Program® studies will provide valid insights into the effectiveness and safety of GH treatment across a diverse and large patient population treated in accordance with real-world clinical practice and following the Good Pharmacoepidemiological Practice and STrengthening the Reporting of OBservational studies in Epidemiology (STROBE) guidelines.


Increased plasma vasopressin in low renin essential hypertension.
Baseline plasma vasopressin concentrations were measured in 48 men (all 50 years old) with decreased plasma renin concentration and untreated, sustained essential hypertension and in 29 healthy normotensive men. Mean hypertensive plasma vasopressin concentration was more than twice as high as the corresponding normotensive level (15.7 +/- 2.2 [SE] vs 7.5 +/- 1.0 pg/ml; p less than 0.001). Plasma renin concentration in the hypertensive group was reduced compared with that in the normotensive group (0.28 +/- 0.04 vs 0.46 +/- 0.06 Goldblatt units X 10(-4)/ml). These differences appeared despite virtually identical serum osmolality, creatinine clearance, and urinary sodium excretion in the two groups. In the first 38 hypertensive subjects, arterial plasma epinephrine concentrations were significantly increased over those of the first 28 control subjects (99 +/- 12 vs 68 +/- 6 pg/ml; p less than 0.025). In contrast to those with low renin essential hypertension, 35 men with normal renin essential hypertension (all 40 years old) had normal plasma vasopressin levels that were not significantly different from those in a comparable normotensive control group (3.7 +/- 0.8 vs 3.5 +/- 0.4 pg/ml). Arterial epinephrine concentrations were not significantly different between normal renin subjects and the control group. After 6 weeks of treatment with the nonselective beta-adrenergic receptor blocker oxprenolol in 11 subjects with low renin hypertension, blood pressure was reduced and the plasma vasopressin concentration fell from 27.6 +/- 6.4 to 13.5 +/- 4.2 pg/ml (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)


[Construction of lentiviral vector of siRNA specific for γ-synuclein and its inhibition effect on colorectal carcinoma cell line SW1116].
To construct a lentiviral vector carrying the γ-synuclein(SNCG) gene and establish a human colorectal carcinoma cell line SW1116 stably expressing this gene, and then investigate the inhibition of the growth and invasion capacity of SW1116 cells. RNA interference fragment was designed according to the SNCG sequence (GenBank: No.NM003087.2), and then SNCG RNAi effective target genes were screened. After the Oligo DNA of target sequences was synthesized, the lentiviral vectors carrying LV-SNCG-RNAi-EGFP (RNAi group) and LV-SNCG-NC-EGFP (NC group) were constructed and packaged to produce lentivirus venom. The supernatants of different virus-producing cells were used to transfect SW1116 cells respectively. Wild SW1116 cells were used as blank control (CON group) EGFP fluorescence was detected by fluorescent microscopy and the differential expression of SNCG mRNA and protein was detected by real-time PCR and Western blot. CCK-8, soft agar assay and Transwell chamber were employed to estimate the inhibiting effect on growth and invasion of SW1116 respectively. Recombinant lentiviral vectors respectively carrying the SNCG-RNAi-EGFP and SNCG-NC-EGFP were successfully constructed and the supernatants of lentivirus could effectively infect SW1116 cells. The titer of the virus carrying LV-SNCG-RNAi-EGFP or LV-SNCG-NC-EGFP was 8×10(8) TU/ml. Real-time PCR and Western blot confirmed that compared with the NC group, SNCG-RNAi group had lower SNCG expression (1.009±0.161 vs. 0.114±0.030, P=0.009), and showed tremendous silencing effect as 76.8%(P<0.05). SNCG protein expression was also significantly reduced (RNAi:12.001±2.884, NC:32.443±4.731, CON:34.308±6.920, P<0.05). After SNCG knockdown, the number of proliferation cells was obviously reduced at 48, 72, 96 and 120 hours respectively(P=0.036). In soft agar assay, clones in RNAi group were smaller[RNAi:(0.582±0.103) mm, NC:(1.863±0.316) mm, CON:(1.749±0.525) mm]. Colony formation rate of RNAi group was down to (17.1±3.5)%, which was significantly lower than (36.5±4.3)% in NC group and (33.8±3.9)% in CON group. In migration test, the number of invasion cell was 37.4±9.3 in RNAi group, which was significantly less than 112.3±8.6 in NC group and 100±0.0 in CON group. Expression of SNCG mRNA and protein plays an important role in the growth and the invasion capacity of SW1116 cells.


Monoclonal B-cell lymphocytosis and chronic lymphocytic leukemia.
A diagnosis of chronic lymphocytic leukemia (CLL) requires a count of over 5000 circulating CLL-phenotype cells per cubic millimeter. Asymptomatic persons with fewer CLL-phenotype cells have monoclonal B-cell lymphocytosis (MBL). The goal of this study was to investigate the relation between MBL and CLL. We investigated 1520 subjects who were 62 to 80 years of age with a normal blood count and 2228 subjects with lymphocytosis (>4000 lymphocytes per cubic millimeter) for the presence of MBL, using flow cytometry. Monoclonal B cells were further characterized by means of cytogenetic and molecular analyses. A representative cohort of 185 subjects with CLL-phenotype MBL and lymphocytosis were monitored for a median of 6.7 years (range, 0.2 to 11.8). Monoclonal CLL-phenotype B cells were detected in 5.1% of subjects (78 of 1520) with a normal blood count and 13.9% (309 of 2228) with lymphocytosis. CLL-phenotype MBL had a frequency of 13q14 deletion and trisomy 12 similar to that of CLL and showed a skewed repertoire of the immunoglobulin heavy variable group (IGHV) genes. Among 185 subjects presenting with lymphocytosis, progressive lymphocytosis occurred in 51 (28%), progressive CLL developed in 28 (15%), and chemotherapy was required in 13 (7%). The absolute B-cell count was the only independent prognostic factor associated with progressive lymphocytosis. During follow-up over a median of 6.7 years, 34% of subjects (62 of 185) died, but only 4 of these deaths were due to CLL. Age above 68 years and hemoglobin level below 12.5 g per deciliter were the only independent prognostic factors for death. The CLL-phenotype cells found in the general population and in subjects with lymphocytosis have features in common with CLL cells. CLL requiring treatment develops in subjects with CLL-phenotype MBL and with lymphocytosis at the rate of 1.1% per year.


Pleurovenous Shunt Placement for the Management of Nonmalignant Pleural Effusion.
Therapeutic thoracentesis is a first-line therapy in the management of patients with medically refractory, nonmalignant pleural effusion. However, when required in short intervals, serial thoracenteses can lead to increased procedure-related complications and negatively impact quality of life. Alternative treatment options vary depending on the etiology of fluid accumulation. Hepatic hydrothorax secondary to cirrhosis is a common cause of medically refractory pleural effusion encountered by interventional radiologists. In select patients in whom surgical pleurodesis, transjugular intrahepatic portosystemic shunt placement, and/or tunneled pleural catheter placement cannot be performed or provide inadequate relief, implantation of a pleurovenous (Denver) shunt may assist in palliation. The Denver shunt system allows decompression of pleural fluid into the central venous circulation by utilizing unidirectional valves and a manually operated subcutaneous pump. Though limited reports have described favorable technical and clinical success, more research is required to determine the safety and efficacy of this procedure. This article discusses pleurovenous shunt placement, postprocedure shunt evaluation, and potential associated complications.


Functional annotation of a full-length Arabidopsis cDNA collection.
Full-length complementary DNAs (cDNAs) are essential for the correct annotation of genomic sequences and for the functional analysis of genes and their products. We isolated 155,144 RIKEN Arabidopsis full-length (RAFL) cDNA clones. The 3'-end expressed sequence tags (ESTs) of 155,144 RAFL cDNAs were clustered into 14,668 nonredundant cDNA groups, about 60% of predicted genes. We also obtained 5' ESTs from 14,034 nonredundant cDNA groups and constructed a promoter database. The sequence database of the RAFL cDNAs is useful for promoter analysis and correct annotation of predicted transcription units and gene products. Furthermore, the full-length cDNAs are useful resources for analyses of the expression profiles, functions, and structures of plant proteins.


Sclerochorioretinal abnormalities in hypercholesterolemic rabbits treated with rosiglitazone.
To evaluate early retinal, choroidal, and scleral abnormalities induced by a hypercholesterolemic diet and the prevention of these abnormalities after oral administration of rosiglitazone in rabbits. Fifty-four New Zealand rabbits were divided into four study groups: control group, normal diet; group 1, hypercholesterolemic diet; group 2, hypercholesterolemic diet associated with daily administration of 3 mg of rosiglitazone from day 14 after beginning the diet; and group 3, hypercholesterolemic diet associated with daily administration of 3 mg of rosiglitazone since the beginning of the experiment. Sclera and choroid underwent histologic and histomorphometric analyses. Retina underwent immunohistochemical analysis with anti-calretinin and anti-glial fibrillary acidic protein (GFAP) antibodies. No abnormalities were observed in the control group. Group 1 had significant increases in scleral and choroidal thicknesses compared with the control group (P < .01) and group 3 (P < .05). Group 1 presented significant increases in immunoreactivity (P < .001) to the anti-calretinin antibody compared with the other groups. Groups 2 and 3 had significant (P < .002) increases in calretinin immunoreactivity compared with the control group. GFAP was negative in all groups. The hypercholesterolemic diet induced early retinal, choroidal, and scleral abnormalities. Rosiglitazone preserved the structural anatomy.


Microbiological-Chemical Sourced Chondroitin Sulfates Protect Neuroblastoma SH-SY5Y Cells against Oxidative Stress and Are Suitable for Hydrogel-Based Controlled Release.
Chondroitin sulfates (CS) are a class of sulfated glycosaminoglycans involved in many biological processes. Several studies reported their protective effect against neurodegenerative conditions like Alzheimer's disease. CS are commonly derived from animal sources, but ethical concerns, the risk of contamination with animal proteins, and the difficulty in controlling the sulfation pattern have prompted research towards non-animal sources. Here we exploited two microbiological-chemical sourced CS (i.e., CS-A,C and CS-A,C,K,L) and Carbopol 974P NF/agarose semi-interpenetrating polymer networks (i.e., P.NaOH.0 and P.Ethanol.0) to set up a release system, and tested the neuroprotective role of released CS against H2O2-induced oxidative stress. After assessing that our CS (1-100 µM) require a 3 h pre-treatment for neuroprotection with SH-SY5Y cells, we evaluated whether the autoclave type (i.e., N- or B-type) affects hydrogel viscoelastic properties. We selected B-type autoclaves and repeated the study after loading CS (1 or 0.1 mg CS/0.5 mL gel). After loading 1 mg CS/0.5 mL gel, we evaluated CS release up to 7 days by 1,9-dimethylmethylene blue (DMMB) assay and verified the neuroprotective role of CS-A,C (1 µM) in the supernatants. We observed that CS-A,C exhibits a broader neuroprotective effect than CS-A,C,K,L. Moreover, sulfation pattern affects not only neuroprotection, but also drug release.


Erratum: Anharmonicity and Universal Response of Linear Carbon Chain Mechanical Properties under Hydrostatic Pressure [Phys. Rev. Lett. 125, 105501 (2020)].
This corrects the article DOI: 10.1103/PhysRevLett.125.105501.


Angiomyolipoma of the liver: immunohistochemical study of a case.
Angiomyolipoma, a not uncommon tumor of the kidney, is rarely encountered in the liver. We report a case of angiomyolipoma of the liver in a 68-year-old woman, in which immunohistochemical study was performed.


Prospective evaluation of allogeneic hematopoietic stem-cell transplantation from matched related and matched unrelated donors in younger adults with high-risk acute myeloid leukemia: German-Austrian trial AMLHD98A.
To assess the impact of allogeneic hematopoietic stem-cell transplantation (HSCT) from matched related donors (MRDs) and matched unrelated donors (MUDs) on outcome in high-risk patients with acute myeloid leukemia (AML) within a prospective multicenter treatment trial. Between 1998 and 2004, 844 patients (median age, 48 years; range, 16 to 62 years) with AML were enrolled onto protocol AMLHD98A that included a risk-adapted treatment strategy. High risk was defined by the presence of unfavorable cytogenetics and/or by no response to induction therapy. Two hundred sixty-seven (32%) of 844 patients were assigned to the high-risk group. Of these 267 patients, 51 patients (19%) achieved complete remission but had adverse cytogenetics, and 216 patients (81%) had no response to induction therapy. Allogeneic HSCT was actually performed in 162 (61%) of 267 high-risk patients, after a median time of 147 days after diagnosis. Graft sources were as follows: MRD (n = 62), MUD (n = 89), haploidentical donor (n = 10), and cord blood (n = 1). The 5-year overall survival rates were 6.5% (95% CI, 3.1% to 13.6%) for patients (n = 105) not proceeding to HSCT and 25.1% (95% CI, 19.1% to 33.0%; from date of transplantation) for patients (n = 162) receiving HSCT. Multivariable analysis including allogeneic HSCT as a time-dependent covariable revealed that allogeneic HSCT significantly improved outcome; there was no difference in outcome between allogeneic HSCT from MRD and MUD. Allogeneic HSCT in younger adults with high-risk AML has a significant beneficial impact on outcome, and allogeneic HSCT from MRD and MUD yields similar results.


Proteomic analysis of mitotic RNA polymerase II reveals novel interactors and association with proteins dysfunctional in disease.
RNA polymerase II (RNAPII) transcribes protein-coding genes in eukaryotes and interacts with factors involved in chromatin remodeling, transcriptional activation, elongation, and RNA processing. Here, we present the isolation of native RNAPII complexes using mild extraction conditions and immunoaffinity purification. RNAPII complexes were extracted from mitotic cells, where they exist dissociated from chromatin. The proteomic content of native complexes in total and size-fractionated extracts was determined using highly sensitive LC-MS/MS. Protein associations with RNAPII were validated by high-resolution immunolocalization experiments in both mitotic cells and in interphase nuclei. Functional assays of transcriptional activity were performed after siRNA-mediated knockdown. We identify >400 RNAPII associated proteins in mitosis, among these previously uncharacterized proteins for which we show roles in transcriptional elongation. We also identify, as novel functional RNAPII interactors, two proteins involved in human disease, ALMS1 and TFG, emphasizing the importance of gene regulation for normal development and physiology.


Prenatal dexamethasone exposure and developmental programming of the ovary of the offspring: a structural study in the rat.
Overexposure to glucocorticoids during fetal development alters fetal organ growth and maturation patterns, which can result in adverse programming outcomes in adulthood. The aim of this study was to determine whether exposure to dexamethasone (Dx) during the fetal period programmed ovary development and function in infant (16-day-old) and peripubertal (38-day-old) female offspring. Pregnant Wistar rats were separated into control and Dx-treated (0.5mg kg-1) groups and were injected with Dx or an equivalent volume of vehicle on Days 16, 17 and 18 of gestation. Ovaries from 16- and 38-day-old female offspring were prepared for histological and stereological examination. The volume of the ovary and the number of primordial and primary follicles were significantly reduced in prenatally Dx-exposed infant and peripubertal female offspring compared with control offspring. The number of multilaminar follicles was decreased in infant female offspring. In peripubertal females, prenatal exposure to Dx increased the number of multilaminar and large follicles of all classes. Because vaginal opening did not occur up to Day 38 postpartum in the Dx-exposed offspring, the absence of ovulation and corpora lutea is confirmation that the onset of puberty had been delayed. We can conclude that overexposure to glucocorticoids early in life programs ovary development, which may affect fertility in adulthood.


Gene for the catalytic subunit of mouse DNA-dependent protein kinase maps to the scid locus.
The gene encoding the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) has been proposed recently as a candidate gene for the mouse severe combined immune deficiency (scid) locus. We have used a partial cDNA clone for human DNA-PKcs to map the mouse homologue using a large interspecific backcross panel. We found that the mouse gene for DNA-PKcs does not recombine with scid, consistent with the hypothesis that scid is a mutation in the mouse gene for DNA-PKcs.


Contributions of transcriptional and post-transcriptional mechanisms to the regulation of c-myc expression in mouse erythroleukemia cells.
Chemically induced differentiation of mouse erythroleukemia (MEL) cells leads to complex changes in c-myc mRNA levels. Within 1-2 hr after the addition of the inducer hexamethylene bisacetamide (HMBA), c-myc mRNA levels decrease 10- to 20-fold and remain low until 12-24 hr, at which time the mRNA reaccumulates to its original level. Thereafter as the cells undergo terminal differentiation, c-myc mRNA again declines to a low level. We have investigated the regulation of these changes by measuring c-myc gene transcription and mRNA turnover. We find that the early rapid decline in c-myc mRNA is due to an increase in the block to elongation of transcription within the c-myc first exon. Effective c-myc transcription is then restored after 2 hr of HMBA treatment to the level present in uninduced cells and is maintained throughout the remainder of the differentiation program. These results demonstrate that, except for the rapid decline in c-myc mRNA immediately following inducer treatment, all subsequent regulation of message levels occurs through post-transcriptional mechanisms. Studies of c-myc mRNA turnover suggest that some post-transcriptional regulation is nuclear.


Molecular dynamics study of hydrated imogolite. 2. Structure and dynamics of confined water.
The behaviour of water confined in an imogolite nanotube was studied by means of molecular dynamics simulations. The results of the study show an important difference between the interaction of water molecules with the internal and external surfaces of the nanotube. The analysis of the density profiles of confined molecules, of their spatial organisation, of the size of molecular clusters, of the lifetime of H-bonds in the system and of dynamical characteristics of molecules permits us to qualify the external imogolite surface as hydrophobic, whereas the internal surface reveals a hydrophilic character.


The effect of physician sleep deprivation on patient safety in perinatal-neonatal medicine.
Recent data indicate that as many as 180,000 patients die each year due to harm suffered as a result of medical care. Between 40 and 60% of these deaths-and the millions of injuries due to medical care-are preventable. In the neonatal intensive care unit (NICU), neonates' size and fragility makes them especially susceptible to serious medical errors, which occur at a far higher rate in this population than elsewhere in hospitals. A growing body of literature demonstrates that the work schedules and sleep deprivation of physicians are important contributors to this epidemic of error. Nowhere is there a higher risk of adverse outcomes due to provider sleep deprivation than in the NICU, where even minor lapses of attention or miscalculations can lead to dire patient outcomes. This review will discuss what is known about how provider sleep deprivation may impact perinatal and neonatal medicine and will discuss ongoing research questions that must be addressed to guide future improvement efforts.


Nitrate transporter NPF7.3/NRT1.5 plays an essential role in regulating phosphate deficiency responses in Arabidopsis.
AtNPF7.3/AtNRT1.5, which is a nitrate transporter that drives root-to-shoot transport of NO3-, is also involved in modulating the response to K+ deprivation in Arabidopsis by affecting root development and K+ transport. However, whether NPF7.3/NRT1.5 functions in regulating plant responses to deficiencies of other nutrients remains unknown. In this study, we found that the expression of AtNPF7.3/AtNRT1.5 was predominant in the roots and was substantially induced by phosphate (Pi) starvation. The atnrt1.5 mutants displayed conspicuously longer primary roots along with a significantly reduced lateral root density under Pi-deficient conditions than did the wild-type plants, and these morphological differences in the roots were eliminated to a certain extent by the ethylene synthesis antagonist Co2+. Further analyses revealed that the expression of important Pi starvation-induced genes, which are directly involved in Pi transport, mobilization and distribution, were significantly higher in the atnrt1.5 mutants than that in the wild-type plants under Pi-starvation conditions; therefore, the atnrt1.5 mutants retained higher tissue Pi concentrations. Taken together, our results suggest that NPF7.3/NRT1.5 is an important component in the regulation of phosphate deficiency responses in Arabidopsis.


Human DNA ligase IV is able to use NAD+ as an alternative adenylation donor for DNA ends ligation.
All the eukaryotic DNA ligases are known to use adenosine triphosphate (ATP) for DNA ligation. Here, we report that human DNA ligase IV, a key enzyme in DNA double-strand break (DSB) repair, is able to use NAD+ as a substrate for double-stranded DNA ligation. In the in vitro ligation assays, we show that the recombinant Ligase IV can use both ATP and NAD+ for DNA ligation. For NAD+-mediated ligation, the BRCA1 C-terminal (BRCT) domain of Ligase IV recognizes NAD+ and facilitates the adenylation of Ligase IV, the first step of ligation. Although XRCC4, the functional partner of Ligase IV, is not required for the NAD+-mediated adenylation, it regulates the transfer of AMP moiety from Ligase IV to the DNA end. Moreover, cancer-associated mutation in the BRCT domain of Ligase IV disrupts the interaction with NAD+, thus abolishes the NAD+-mediated adenylation of Ligase IV and DSB ligation. Disrupting the NAD+ recognition site in the BRCT domain impairs non-homologous end joining (NHEJ) in cell. Taken together, our study reveals that in addition to ATP, Ligase IV may use NAD+ as an alternative adenylation donor for NHEJ repair and maintaining genomic stability.


Genetics of melanoma susceptibility.
Both genetic and environmental factors confer a significantly increased risk for cutaneous melanoma. This review discusses hereditary predisposition to the disease, focusing on the high-penetrance candidate genes INK4A/ARF and CDK4, and on pathogenetic mechanisms of mutations in those genes. As known mutations account for approximately 25 to 40% of melanoma families reported to date, it is clear that other melanoma genes and other mechanisms underlying predisposition remain to be discovered. Low penetrance susceptibility genes such as melanocortin 1 receptor and their modifying effect, also in concert with UV radiation, are likely to be implicated. Recent reports on a new candidate locus on chromosome 1p22 and somatic mutations in genes of the RAS-RAF-ERK signalling pathway raise interesting questions for further investigation.


Development of a new software and test setup for analyzing hVOR in very young children by vHIT.
Earlier work revealed that vHIT examination is often difficult to perform on very young children. In particular, the calibration of the system can be difficult, as active cooperation of the patient is required. Additionally, the patient must be able to follow the examiner's instructions, which is challenging for very young children. Therefore, the aim of the present study was to develop and validate a new, software-based approach enabling vHIT testing of young children and infants. Six patients (3 boys and 3 girls) aged 5-36 months were included in a prospective, monocentric study between January 2015 and August 2015.The newly developed intuitive software enabled calibration of the eye position signal with the subjects fixating on animated animal graphics which were projected on a screen. Testing ten healthy adults validated this new calibration and measurement method. After calibration, a vHIT goggle (EyeSeeCam ©) was used to perform head impulses in the horizontal plane while the patient was watching a movie sitting on their parent's lap or in a baby chair. At least 15 impulses to each side were obtained and the occurrence of refixation saccades was analyzed. All tests were performed by one of two experienced examiners. The new calibration method and modified test setup provided reproducible results for all patients tested. An increased incidence of artifacts was not observed. In 2 patients, more than one test was needed. None of the included children showed catch-up overt or catch-up covert saccades. There was no gain reduction of more than two standard deviations as compared to the normative results published in the literature on vHIT examinations of children. The proposed protocol allows vHIT testing in very young children and infants (aged 5 months to 3 years). The study emphasizes that vHIT is an easy and sensitive screening tool to evaluate vestibular function in children and should be used as the gold standard in pediatric vestibular assessment.


Association of genetic variation in pharmacodynamic factors with methadone dose required for effective treatment of opioid addiction.
The interindividual variability in the dose required for effective methadone maintenance treatment (MMT) for opioid addiction may be influenced in part by genetic variations in genes encoding pharmacodynamic factors of methadone. This study was conducted to identify some of these variants. This study focused on 11 genes encoding components of the opioidergic (OPRM1, POMC and ARRB2), the dopaminergic (ANKK1 and DRD2) and the glutamatergic pathways (GRIN1 and GRIN2A), as well as the neurotrophin system (NGFB, BDNF, NTRK1 and NTRK2). The study includes 227 Israeli patients undergoing stable MMT. Out of the 110 variants analyzed, 12 SNPs (in BDNF, NTRK2, OPRM1, DRD2 and ANKK1) were associated with methadone dose (nominal p < 0.05). Of these SNPs, ANKK1 rs7118900 and DRD2 rs2283265 are known to affect gene expression. Logistic regression of five representative SNPs discriminated between individuals requiring a methadone dose of >120 mg/day and <120 mg/day (p = 0.019), and showed moderate sensitivity and specificity (AUC of 0.63 in receiver operating characteristic analysis). This data should stimulate further research on the potential influence and clinical significance of these variants on MMT.