Amphiphilic peptide carrier for the combined delivery of curcumin and plasmid DNA into the lungs.
In this study, the R7L10 peptide, which is composed of a 7-arginine stretch and a 10-leucine stretch, was evaluated as a carrier for the combined delivery of curcumin and plasmid DNA (pDNA) into the lungs. Curcumin is a natural product with anti-inflammatory and anti-tumor effects. Curcumin-loaded R7L10 (R7L10-curucmin) was prepared by an oil-in-water (O/W) emulsion/solvent evaporation method. In vitro transfection showed that R7L10-curcumin had higher transfection efficiency than R7L10. Although R7L10-curcumin had lower transfection efficiency than polyethylenimine (25 kDa, PEI25k) and lipofectamine, R7L10-curcumin had lower cytotoxicity. In gel retardation assays and heparin competition assays, R7L10-curcumin formed a more stable complex with pDNA than R7L10. The intracellular curcumin delivery efficiency of R7L10-curcumin was higher than that of curcumin only. Furthermore, R7L10-curcumin more efficiently decreased TNF-α level in lipopolysaccharide (LPS)-activated Raw264.7 macrophage cells than curcumin only. For in vivo evaluation, pDNA/R7L10-curcumin complexes were administered into mouse lungs by intratracheal instillation. The results revealed that R7L10-curcumin delivered pDNA more efficiently than R7L10, poly-L-lysine (PLL), or PEI25k. In addition, R7L10-curcumin decreased TNF-α level in lung tissues in an acute lung injury mouse model. In contrast to PEI25k, R7L10-curcumin did not show liver toxicity after intravenous injection. These results suggest that R7L10-curcumin is a useful carrier for the combined delivery of curcumin and pDNA into the lungs.


Genetic Association of Butyrylcholinesterase with Major Depressive Disorder.
Major depressive disorder (MDD) is characterized as clinical depression, which primarily affects the mood and behaviour of an individual. In the present study butyrylcholinesterase (BChE), a co-regulatory cholinergic neurotransmitter enzyme implicated in several putative neuronal and non-neuronal physiological roles was investigated for its role in MDD. Eighty MDD patients and sixty-one healthy controls were recruited for the study. BChE activity was measured by Ellman's method using serum while DNA samples of the patients were genotyped for BCHE polymorphisms rs3495 (c.*189G > A) and rs1803274 (c.1699G > A) by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and tetra-primer Amplification Refractory Mutation System- polymerase chain reaction (ARMS-PCR). The genotyping was further validated by Sanger Sequencing. Biochemical estimation of serum BChE levels revealed a statistically significant decrease of enzyme activity in MDD patients (69.96) as compared to healthy controls (90.97), which was independent of age and gender. BCHE single nucleotide polymorphism rs1803274 genotype GA was found to be associated with the disease under a dominant model (OR 2.32; 95% CI 1.09-4.96; p value = 0.025). Furthermore, risk allele-A frequency was higher in cases (p value = 0.013) than control. Carriers of rs1803274 GA genotype showed reduced mean BChE activity than wild-type allele GG homozygotes (p value = 0.040). Gender-based analysis revealed a protective role of rs3495 in females (χ2 = 6.87, p value = 0.032, RM: OR 0.173, CI = 0.043-0.699 (p value = 0.017). In addition, rs1803274 risk allele-A was observed to be significantly higher in males (χ2 = 4.258, p value = 0.039). In conclusion, the present study is indicative of a role of BChE in the pathophysiology of MDD where genetic polymorphisms were observed to effect BChE activity. Further replication studies in different ethnicities are recommended to validate the current observations.


Mechanically inspired laser scribing of thin flexible glass.
Laser cutting of thin glass (<100 μm) has proven problematic. We describe an alternative laser scribing method that utilizes surface stress raisers. An ultrashort laser source is used to precisely pattern a plurality of aligned elliptical recesses on the glass. The apex of an ellipse concentrates applied tensile stresses. Depending on the elliptical dimensions, the stress concentration factor can be several tens of magnitude. The orientation of the ellipses defines a preferred scribing path. Tensile stress is applied orthogonally to the path and causes mode I fracture. The resulting scribe is of higher quality and strength than are possible with a full body laser cut. The optical setup is simple, low in cost, and compatible with future roll-to-roll manufacturing. The stress field around a stress raiser was analyzed using finite element method analysis. Consequently, the stress raiser process offers an alternative to other processes which employ high numerical aperture optics for thin glass scribing.


Keeping Superprotonic Conductivity over a Wide Temperature Region via Sulfate Hopping Sites-Decorated Zirconium-Oxo Clusters.
Metal-oxo clusters have emerged as advanced proton conductors with well-defined and tunable structures. Nevertheless, the exploitation of metal-oxo clusters with high and stable proton conductivity over a relatively wide temperature range still remains a great challenge. Herein, three sulfate groups decorated zirconium-oxo clusters (Zr6 , Zr18 , and Zr70 ) as proton conductors are reported, which exhibit ultrahigh bulk proton conductivities of 1.71 × 10-1 , 2.01 × 10-2 , and 3.73 × 10-2 S cm-1 under 70 °C and 98% relative humidity (RH), respectively. Remarkably, Zr6 and Zr70 with multiple sulfate groups as proton hopping sites show ultralow activation energies of 0.22 and 0.18 eV, respectively, and stable bulk conductivities of >10-2 S cm-1 between 30 and 70 °C at 98% RH. Moreover, a time-dependent proton conductivity test reveals that the best performing Zr6 can maintain high proton conductivity up to 15 h with negligible loss at 70 °C and 98% RH, representing one of the best crystalline cluster-based proton conducting materials.


MERRF/MELAS overlap syndrome in a family with A3243G mtDNA mutation.
Four members of a family were found to carry the A3243G mtDNA mutation. Clinical features varied from typical MELAS to myoclonic epilepsy to simple deafness without neurological signs. Several other members of the family had symptoms consistent with a mitochondrial disease. Muscle biopsy in 3 of the 4 patients showed the most prominent mitochondrial alterations with partial deficiency of cytochrome c oxidase in the case with the mildest phenotype. Mitochondrial DNA analysis detected a variable percentage of A3243G mutation, roughly correlating with the phenotype. The interesting feature of the family lies in the great intrafamilial variability of the severity of clinical expression, encompassing MELAS and MERRF features, associated with the A3243G mtDNA mutation. A search for the most common mtDNA mutations is recommended in all patients featuring incomplete MELAS or MERRF syndromes and in all familial cases presenting minimal clinical signs.


Mortality Burden of Cardiovascular Disease Attributable to Ozone in China: 2019 vs 2050.
Due to a combination of lifestyle risk factors, the burden of cardiovascular disease (CVD) has been increasing in China, affecting an estimated 330 million people. Environmental risk factors can exacerbate these risks or independently contribute to CVD. Ozone is an overlooked and invisible risk factor, and it plays a significant role in the development of CVD. Our study provides a novel quantification of the ozone-attributable CVD mortality burden based on daily maximum 8-h average ozone concentration during May to October (6mDMA8) in Chinese adults in 2050, projected under Shared Socioeconomic Pathways 585 and 126, and using the updated WHO air quality guideline level. The study also considers the contributions made by changes in ozone exposure, population aging, population size, and baseline death rates of CVD between 2019 and 2050. While adopting a sustainable and green pathway (SSP 126) can reduce the projected magnitude of premature CVD deaths to 359,200 in 2050, it may not be sufficient to reduce the CVD mortality burden significantly. Therefore, it is crucial to implement strategies for stricter ozone control and reducing the baseline death rate of CVD to mitigate the impacts of ozone on Chinese adults.


HLA-DQA1 and DQB1 alleles in French and Algerian type 1 diabetic subjects.
Some alleles of the HLA-DQB1 and DQA1 loci are preferentially associated with susceptibility to type 1 (insulin-dependent) diabetes mellitus (IDDM). Analysis of the HLA-DQ genetic profile may therefore become important for the screening of subjects at risk of IDDM. However ethnic variations in the genetic profile can occur and require background knowledge of the HLA-DQ allelic distribution before screening campaigns. In the present work, HLA-DQA1 and DQB1 genes have been analyzed, after PCR amplification of the genomic DNA, in French and Algerian control subjects (a total of 148) and diabetic patients (a total of 107). Allelic distributions have been investigated in view of a) possible inter-ethnic differences; b) identification of risk and protective alleles and c) the prevalence of DQB1 aspartate 57 negative and DQA1 arginine 52 positive alleles in control and diabetic groups. The DQB1 allelic distribution was similar in both control groups; alleles negative for aspartate at position 57 were 48% in French and 50% in Algerian. In both diabetic groups, the prevalence of alleles negative for aspartate at position 57 was significantly higher: 91% (French) and 81% (Algerian) (p less than 0.001). A majority of patients were homozygote for DQB1 Asp 57 negativity: 83% (French) and 63% (Algerian). The highest relative risk was associated with HLA-DQB1 0201/0302 heterozygosity. The HLA-DQA1 allelic distribution was also similar in French and Algerian controls. Alleles positive for arginine (ARG+) at position 52 were 50% (French) and 57% (Algerian) of controls. In both diabetic groups the prevalence of alleles positive for arginine at position 52 was significantly higher: 78% (French) and 84% (Algerian).(ABSTRACT TRUNCATED AT 250 WORDS)


Nutrient Channels Aid the Growth of Articular Surface-Sized Engineered Cartilage Constructs.
Symptomatic osteoarthritic lesions span large regions of joint surfaces and the ability to engineer cartilage constructs at clinically relevant sizes would be highly desirable. We previously demonstrated that nutrient transport limitations can be mitigated by the introduction of channels in 10 mm diameter cartilage constructs. In this study, we scaled up our previous system to cast and cultivate 40 mm diameter constructs (2.3 mm overall thickness); 4 mm diameter and channeled 10 mm diameter constructs were studied for comparison. Furthermore, to assess whether prior results using primary bovine cells are applicable for passaged cells-a more clinically realistic scenario-we cast constructs of each size with primary or twice-passaged cells. Constructs were assessed mechanically for equilibrium compressive Young's modulus (EY), dynamic modulus at 0.01 Hz (G*), and friction coefficient (μ); they were also assessed biochemically, histologically, and immunohistochemically for glycosaminoglycan (GAG) and collagen contents. By maintaining open channels, we successfully cultured robust constructs the size of entire human articular cartilage layers (growing to ∼52 mm in diameter, 4 mm thick, mass of 8 g by day 56), representing a 100-fold increase in scale over our 4 mm diameter constructs, without compromising their functional properties. Large constructs reached EY of up to 623 kPa and GAG contents up to 8.9%/ww (% of wet weight), both within native cartilage ranges, had G* >2 MPa, and up to 3.5%/ww collagen. Constructs also exhibited some of the lowest μ reported for engineered cartilage (0.06-0.11). Passaged cells produced tissue of lower quality, but still exhibited native EY and GAG content, similar to their smaller controls. The constructs produced in this study are, to our knowledge, the largest engineered cartilage constructs to date which possess native EY and GAG, and are a testament to the effectiveness of nutrient channels in overcoming transport limitations in cartilage tissue engineering.


Ocular findings, surgery details and outcomes in proliferative diabetic retinopathy patients with chronic kidney disease.
We investigated the influence of impaired renal function on fundus characteristics, pars-plana vitrectomy (PPV) details, and outcomes in patients with proliferative diabetic retinopathy (PDR). A retrospective cohort study. We investigated a consecutive series of PDR patients who underwent PPV. The diabetic complications, previous photocoagulation, intravitreal injections before PPV, ocular findings during PPV, surgical details, short-term visual outcome and post-PPV complications were recorded and compared between patients with and without impaired renal function. 149 patients had normal renal function (67.7%), and 71 (32.3%) patients had impaired renal function; 85.4% of patients were identified with chronic kidney disease (CKD) during the preoperative assessment. Impaired renal function was related to hypertension (3.40[1.58-7.29], p = 0.002), incomplete pan-retinal photocoagulation (PRP) (3.18[1.50-6.72], p = 0.002), severe fibrovascular membrane (8.19[3.43-19.54], p<0.001), and extensive retinal vascular closure (3.40[1.54-7.52], p = 0.002). There was a more frequent occurrence of severe intraoperative bleeding (56.3%, 32.2%, p = 0.001) and a higher percentage of intraocular subretinal fluid drainage (45.1%, 22.1%, p = 0.008) in patients with impaired renal function. The percentage of patients whose visual acuity (VA) increased was similar between the two groups (42.3%, 54.4%, p = 0.34). In PDR patients, screening for CKD was required before PPV. PDR patients with impaired renal function tended to have more severe ischemic retinal conditions. Comparable PPV outcomes could be obtained in patients with and without impaired renal damage.


[Adult progeria (Werner's syndrome)].
A 48-year-old man was referred with progressive claudication and nocturnal pain at rest in the right foot. During the last few years exercise-induced pain, ulcerations and necroses had developed in both legs without any evidence of relevant macroangiopathies. The patient showed marked morphological changes. He had bilateral cataracts since aged 35 years. At 40 years he had undergone a transurethral resection of a bladder carcinoma. One of his brothers showed similar morphological changes, but they were absent in his parents and three siblings. Radiology of the right foot revealed extensive para-osseous soft-tissue calcifications. Direct right femoral angiography demonstrated distal occlusions of the anterior and posterior tibial arteries as well as subtotal occlusion of the fibular artery. Routine laboratory tests were normal, as were all measured antibody titres. The peripheral vascular disease, stage IV, of the right leg were presumably caused by adult progeria (Werner's syndrome). Percutaneous transluminal angioplasty (PTAP) of the distal fibular and, later, of the posterior tibial artery was performed, and prostacyclins, antibiotics and platelet-aggregation inhibitors were administered. Although the PTAP was technically successful, the state of the right foot remained critical, due to the existing microangiopathy and the impaired proliferation and healing capacity of the tissues as part of the adult progeria. The diagnosis of adult progeria is usually made because of the patients' characteristic morphology and typical concomitant diseases. The average age of patients at diagnosis is 37 years. The syndrome is caused by a helicase defect. This enzyme group unwinds double-helix RNA and DNA. Because the syndrome is rare and gene analysis is complex, it is unlikely that a commercial screening test will become available.


Emotional and sensory dysregulation as a possible missing link in attention deficit hyperactivity disorder: A review.
Attention Deficit Hyperactivity Disorder (ADHD) is a common developmental disorder affecting 5-7% of adults and children. We surveyed the literature to examine ADHD through three pillars: developmental characteristics, symptomatology, and treatment strategies. Firstly, in terms of developmental characterstics, early life stress may increase the risk of developing ADHD symptoms according to animal models' research. Secondly, the current core symptoms of ADHD are comprised of inattention, hyperactivity, and impulsivity. However, the up-to-date literature indicates individuals with ADHD experience emotional and sensory dysregulation as well, which early-life stress may also increase the risk of. Finally, we discuss the therapeutic benefits of methylphenidate on both the current core ADHD symptoms and the sensory and emotional dysregulation found in those with ADHD. In summation, we surveyed the recent literature to analyze (i) the potential role of early-life stress in ADHD development, (ii) the involvement of emotional and sensory dysregulation in ADHD symptomatology and finally, (iii) the therapeutic intervention with methylphenidate, aiming to reduce the potential effect of early life stress in ADHD, and mainly emotional and sensory dysregulation. The apparent but currently less recognized additional symptoms of emotional and sensory dysregulation in ADHD call for further investigation of these possible causes and thus increasing treatments efficacy in individuals with ADHD.


Liver disease with unknown etiology - have you ruled out alpha-1 antitrypsin deficiency?
Although a less well-known consequence of alpha-1 antitrypsin deficiency (AATD) liver disease is the second leading cause of death among patients with the condition. The alpha-1 antitrypsin (AAT) protein is produced by hepatocytes within the liver, which retain pathological variants of AAT instead of secreting the proteinase inhibitor into the systemic circulation. This intracellular retention is caused by inefficient folding and polymerization of mutant AAT and the accumulation of these AAT aggregates leads to diverse manifestations of liver disease, which can present differently in both children and adults. The progression from hepatocyte apoptosis to liver inflammation, fibrosis and cirrhosis, and liver failure is still not fully understood, but in older patients, liver disease can surpass lung disease as the principal cause of death. Liver function tests (LFTs) can measure plasma levels of liver enzymes to assess liver function but require careful interpretation. Non-invasive tests are being developed that can detect early liver disease, but liver biopsy is still the gold standard for assessing liver fibrosis once abnormal LFTs have been detected in a patient. Currently, there is no licensed treatment for AATD-related liver disease (intravenous AAT therapy is not indicated for this purpose), but liver transplantation is associated with positive outcomes and may even slow emphysema progression. Therefore, new strategies are being developed to address treatment of AATD-related liver disease, such as accelerating degradation of mutant AAT and assisting hepatocytes in the folding and secretion of mutant AAT, but these approaches remain at early stages of development.


Practical experience with databases for congenital heart disease: a registry versus an academic database.
Increasingly, pooled data from multiple institutions are the source of published clinical results. A computerized database program is essential to compile and analyze clinical experience. The scope of data collection defines a database. Two types of databases, the registry and academic, are compared. In a registry database, some of the data are collected on all patients. The resources dedicated to data collection and entry are the practical limit to the extent of information in the database. The agreement on nomenclature for surgical diagnosis and procedure codes of congenital heart disease has paved the way for the development of a multi-institutional registry database. The registry database could provide a standard of care reference for early results after congenital heart surgery. The practical difficulty of data collection is obviated by limiting information to a basic minimum dataset. The academic database, in which all of the data are collected for a defined subset of patients, is designed to investigate a specific population of patients to generate new knowledge. It contains sufficient data to allow sophisticated statistical analysis to clarify the determinants of good and poor outcome, including early, mid- and long-term follow-up information. Multi-institutional pooling of detailed information derived from academic databases will be of increasing importance in generating new knowledge to foster improved therapy for patients with congenital heart disease.


Potassium hydroxide 5% for the treatment of molluscum contagiosum.
Molluscum contagiosum is a common reason for consultation in primary care. The condition is normally benign and self-limiting1 and the standard advice is to wait for the lesions to resolve spontaneously.2 Recently, potassium hydroxide 5% (MolluDab-Alliance Pharmaceuticals Limited) has been marketed in the UK for the treatment of the condition.3 It is sold as a medical device rather than a licensed medicinal product. Here we consider the evidence for potassium hydroxide 5% in the management of molluscum contagiosum.


Bariatric surgery for type 2 diabetes: to whom and when?
Achieving clinical and biochemical targets in type 2 diabetes mellitus (diabetes) is fundamental to preventing diabetic microvascular and macrovascular complications. These clinical imperatives are made more challenging in the setting of obesity. This review examines the evidence for bariatric surgery as a means of optimizing diabetes care, addressing diabetes resolution, glucose and lipid metabolism and cardiac effects. Mechanistic evidence of diabetes amelioration is summarized. The question of when bariatric surgery should be offered in the course of obesity-associated diabetes is discussed. Current guidelines and specialist society recommendations for bariatric surgery in diabetes are summarized.


Granulomatous Lobular Mastitis Associated with Mycobacterium abscessus in South China: A Case Report and Review of the Literature.
Mycobacteria, which are known as rapidly growing bacteria, are pathogens that are responsible for cutaneous or subcutaneous infections that especially occur after injection, trauma, or surgery. In this report, we describe a species of Mycobacterium abscessus that was isolated from a breast abscess in a patient who was previously diagnosed with granulomatous lobular mastitis (GLM). This current case is the first ever presented case of GLM associated with M. abscessus documented in South China. The case presentation highlights the role of M. abscessus in GLM. The association of M. abscessus and GLM is discussed and a summary of breast infection due to Mycobacteria is given.


PCSK9 R46L, lower LDL, and cardiovascular disease risk in familial hypercholesterolemia: a cross-sectional cohort study.
Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a downregulator of the low density lipoprotein receptor. The aims of this cross-sectional cohort-study were to examine whether the PCSK9 R46L loss of function variant found in a cohort of familial hypercholesterolemia (FH) patients was associated with lower low density lipoprotein cholesterol, lower frequency of xanthomata, and cardiovascular risk. We studied FH patients attending the IRCM (Institut de Recherches Cliniques de Montréal) Lipid Clinic and whose DNA genotyping was positive for a low density lipoprotein receptor mutation. The presence of the PCSK9 loss of function R46L missense variant was determined among a cohort of 582 FH patients by genotyping. Frequency of the R46L variant was 3%. Carriers had significantly lower low density lipoprotein cholesterol (11%, P=0.002), total cholesterol (9%, P=0.007), apolipoprotein B (10%, P=0.037), and non-high density lipoprotein (12%, P<0.001) concentrations compared with noncarriers. Furthermore, R46L carriers showed a decreased average number of xanthoma per individual compared with noncarriers (0.33 and 0.76, respectively; P<0.001). Importantly, the R46L genetic variant was associated with a significant 86% lower odd of presenting a cardiovascular event (odds ratio, 0.14; 95% confidence interval, 0.032-0.63; P=0.001). Even though the R46L variant was present in 3% of our FH population, carriers of this polymorphism showed attenuated effect of the low density lipoprotein receptor mutation on parameters, such as low density lipoprotein cholesterol, apolipoprotein B, total cholesterol, and non-high density lipoprotein. More importantly, this mutation is associated with a significant lower risk of cardiovascular disease compared with noncarriers. It is therefore likely that targeting PCSK9 in FH patients with novel anti-PCSK9 therapies will be useful in reducing cardiovascular risk in affected subjects.


A suppressor analysis of residues involved in cation transport in the lactose permease: identification of a coupling sensor.
Four amino acids critical for lactose permease function were altered using site-directed mutagenesis. The resulting Quad mutant (E269Q/R302L/H322Q/E325Q) was expressed at 60% of wild-type levels but found to have negligible transport activity. The Quad mutant was used as a parental strain to isolate suppressors that regained the ability to ferment the alpha-galactoside melibiose. Six different suppressors were identified involving five discrete amino acid changes and one amino acid deletion (Q60L, V229G, Y236D, S306L, K319N and DeltaI298). All of the suppressors transported alpha-galactosides at substantial rates. In addition, the Q60L, DeltaI298 and K319N suppressors regained a small but detectable amount of lactose transport. Assays of sugar-driven cation transport showed that both the Q60L and K319N suppressors couple the influx of melibiose with cations (H(+) or H(3)O(+)). Taken together, the data show that the cation-binding domain in the lactose permease is not a fixed structure as proposed in previous models. Rather, the data are consistent with a model in which several ionizable residues form a dynamic coupling sensor that also may interact directly with the cation and lactose.


The Next Generation of KRAS Targeting: Reasons for Excitement and Concern.
The development of selective KRASG12C inhibitors that directly inhibit KRAS, an oncogene historically thought to be "undruggable," represents a watershed moment in oncology and developmental therapeutics. Now, as KRAS-targeted therapy moves into its second phase, there is significant excitement and anticipation for durable disease control in tumor types where options remain limited, with clinical trials testing combination therapies, indirect pan-RAS/MAP kinase pathway inhibitors, and active-state RAS(on) inhibitors. However, there is also reason for caution regarding the safety and tolerability of expanded RAS inhibition. This is evidenced by the intolerability of some combination therapies with selective KRASG12C inhibitors and foreshadowed by prior failures of combination therapies in other oncogene-driven tumors. Herein, we review the landscape of and outlook for KRAS-targeted therapies. We specifically focus upon strategies to combat resistance to KRAS-targeted therapies, and discuss the possibility of off-target or unanticipated on-target effects that may limit clinical use.


Absence of correlations in the QCD dirac spectrum at high temperature.
I propose a simple model of the distribution of the small eigenvalues of the QCD Dirac operator well above the finite temperature phase transition where chiral symmetry is restored and the spectral density at zero vanishes. Assuming the absence of correlations between different regions of the low-lying spectrum I derive analytic formulas for the distribution of the first two eigenvalues. I find good agreement with data obtained using the overlap Dirac operator in quenched SU(2) lattice simulations. This suggests that if chiral symmetry is restored spectral correlations are not important and all the statistical properties of the spectrum are encoded in the spectral density.


The first Japanese nationwide multicenter study of BRCA mutation testing in ovarian cancer: CHARacterizing the cross-sectionaL approach to Ovarian cancer geneTic TEsting of BRCA (CHARLOTTE).
BRCA gene mutations are associated with hereditary ovarian cancer. BRCA plays a key role in genome integrity, and mutations result in an increased risk for ovarian cancer. Although various guidelines recommend BRCA testing in patients with ovarian cancer, data on germline BRCA (gBRCA) mutation frequency in ovarian cancer in Japan are scarce. This study aimed to determine gBRCA1/2 mutations in Japanese patients with ovarian cancer, stratified by clinicopathological characteristics, and to assess patients' satisfaction with pre-test genetic counseling. The CHARLOTTE study (CHARacterizing the cross-sectionaL approach to Ovarian cancer: geneTic TEsting of BRCA; UMIN000025597) is the first large multicenter epidemiological survey of Japanese women, aged ≥20, with newly diagnosed ovarian cancer (epithelial, primary peritoneal, or fallopian tube cancer), with histologically confirmed specimens. Patients were enrolled sequentially and underwent pre-test genetic counseling for BRCA testing. Blood samples were centrally tested for the presence or absence of known gBRCA mutations. A questionnaire was used to assess patient satisfaction with pre-test genetic counseling. A total of 634 patients with a mean age of 56.9 years were included. Most patients (84.2%) had epithelial ovarian cancer, and 51.1% had FIGO stage III-IV cancer. Nearly all patients (99.5%) received genetic counseling before the BRCA testing, either by an obstetrician-gynecologist (42.0%) or a clinical geneticist (42.0%). The overall prevalence of gBRCA1/2 mutations was 14.7% (93/634), with gBRCA1 mutations (9.9%) more common than gBRCA2 mutations (4.7%). High-grade serous carcinoma showed a prevalence of gBRCA mutations of 28.5%. Most patients were satisfied with pre-test counseling, irrespective of the service provider's professional position. Patients with high-grade serous carcinoma and family history of ovarian cancer had a slightly higher prevalence of gBRCA mutations, but none of the subgroups had considerably high gBRCA mutation prevalence. These data suggest that gBRCA testing should be carried out in all patients with ovarian cancer.


[Motivational Goals and Couple Relationship Quality in a Sample of Individuals with Infertility].
3,5-16,6% of couples worldwide with childwish suffer from infertility, and about 20% thereof show clinically relevant psychological symptoms. Until now it remained unclear, up to which extent motivational factors are associated with psychological distress when suffering from infertility. The goal of this study was to assess individual motivational factors in a sample of infertile people, and to explore their effect on psychological distress. Additionally, the role of couple relationship quality as a dyadic predictor was to be examined. We assessed psychological distress, specificity of motivational goals and their achievement, and couple relationship quality in a sample of 123 infertile subjects by online survey. About half of the subjects indicated significant levels of psychological distress. Compared to psychopathologically distressed samples (without explicit infertility problems), infertile subjects specified less approach motivation. Furthermore, low levels of approach motivation were associated with low levels of distress. This applied specifically to the need for control and self-worth. Motivational factors explained 30% of variance in distress, whereas couple relationship quality, specifically the prevalence of arguing, explained merely 4%. These findings show that infertility is associated with distinct motivational patterns, and that these may be suitable to predict psychological distress.


Short-term use of ceftriaxone sodium leads to intestinal barrier disruption and ultrastructural changes of kidney in SD rats.
Antibiotic treatments are known to disturb gut microbiota, but their effects on the mucosal barrier and extraintestinal diseases are rarely discussed. The aim of this study was to evaluate and visualize the impact of antibiotics on colonic mucus and the microbial community, and to assess whether intestinal dysbacteriosis is involved in the pathogenesis and progression of extraintestinal diseases in vivo. Twenty-one SD rats were randomly assigned into three groups followed by different experimental treatments. The albumin-creatinine ratio, urinary protein and occult blood semi-quantified test were tested. Fecal samples were collected at different time points (0,4, and 12 weeks) for 16S rRNA gene sequencing. Colon and kidney specimens were examined using light microscopy and transmission electron microscopy (TEM) to identify morphological changes. Ceftriaxone intervention for one week did not cause any symptoms of diarrhea or weight loss, but the alpha and beta diversities of gut microbiota decreased quickly and significantly, a lower Firmicutes/Bacteroidetes (F/B) ratio was observed. At week 12, although the alpha and beta diversities increased to a level similar to that of the control (CON) group, LEfSe analysis indicated that the microbial community composition still differed significantly in each group. In addition, KEGG metabolic prediction revealed different metabolic functions in each group. TEM examination of colon revealed that dramatic morphological changes were observed in the ceftriaxone (Cef) group, wherein microvilli were misaligned and shortened significantly and morphologically intact bacteria were seen on the epithelial cell surface. TEM examination of kidneys from the Cef group showed characteristic glomerular changes in the form of widely irregularly thickened glomerular basement membrane (GBM) and foot process fusion or effacement; mild thickening of the GBM and foot process fusion was detected when ceftriaxone and Resatorvid (TAK242, an inhibitor of TLR4 signaling) are used together in the ceftriaxone + TAK242 (TAK) group. Short-term use of ceftriaxone induced dynamic changes of gut microbiota and lead to intestinal barrier disruption and ultrastructural changes of kidneys in the SD rats. Moreover, interference with the TLR4-dependent signaling pathway can alleviate the damage to the intestinal barrier and kidney.


[Be careful in using flecainide by hemodialyzed patients].
Cardiac arrhythmias are frequent by hemodialyzed patients and may lead to the prescription of antiarrhythmic drugs among which flecainide. However there is for this latter a wide interindividual variability in the dose-plasma concentration relationship because of a genetically controlled polymorphic metabolism. That explains why it is not possible to adjust dose only to renal clearance. It becomes even more difficult at hemodialysis stage, especially if hepatic insufficiency and/or certain medications join to it. So this antiarrhythmic drug has to be cautiously initiated and plasma concentrations have to be carefully monitored to avoid side effects, sometimes crippling as in our observation, sometimes as cardiac complications.


The Outcome of Large B-Cell Lymphoma Evolving in a Hematopoietic Stem Cell Transplant Patient During Treatment of Chronic Graft-Versus-Host Disease.
Immunosuppression is a risk factor for the development of posttransplant lymphoproliferative disorder. This type of malignancy developed in an immunocompromised man. The patient presented with focal convulsion starting from right face and right arm followed by two generalized convulsions with one minute interval. Diagnosis was made by computed tomographic (CT) scan of the cranium following oral and intravenous administration of contrast dyes which revealedmass leisons in the frontotemporal lobe and by stereotaxic biopsy. Tissue sections showed a malignant tumor cell infiltration with large areas of necrosis and many mitoses. Many tumor cells were positive for CD 20 and CD 10. These findings were consistent with large B-cell lymphoma. Central nervous system radiation with a dose of 56 Gy was given with clinical and radiological improvement. The patient died due to multiorgan failure. Finally, the immunocompromised patients should be closely followed for the development of lymphoproliferative disorder.


Evaluating the Impact of Dictionary Updates on Automatic Annotations Based on Clinical NLP Systems.
Concept encoding, which maps text spans to concepts in standard terminologies, is a critical component in clinical natural language processing (NLP) systems to allow semantic interoperability with other clinical applications. A majority of clinical NLP systems adopt dictionary or lexicon based approaches and the performance of concept encoding is often evaluated using a human created gold standard generated with reference to the most up-to-date standard terminologies available at the time of gold standard creation. With the advance of medical science, standard terminologies or dictionaries can evolve. However, it remains unknown whether the dictionary updates will impact the performance of concept encoding. In this study, we evaluated the annotation performance of two clinical NLP systems, cTAKES and MedXN based on updated dictionaries to gain further insights. Specifically, we compared the automatic annotation results with previously manually generated gold standards. The results of our study demonstrate the annotation changes based on dictionary updates in clinical NLP systems and that it is necessary to do temporal management for gold standards, which raises the need for appropriate terminology management tools for back version compatibility to update gold standards.


Prolactin, progesterone, and dexamethasone coordinately and adversely regulate glucokinase and cAMP/PDE cascades in MIN6 beta-cells.
Islet cells undergo major changes in structure and function to meet the demand for increased insulin secretion during pregnancy, but the nature of the hormonal interactions and signaling events is incompletely understood. Here, we used the glucose-responsive MIN6 beta-cell line treated with prolactin (PRL), progesterone (PRG), and dexamethasone (DEX, a synthetic glucocorticoid), all elevated during late pregnancy, to study their effects on mechanisms of insulin secretion. DEX alone or combined with PRL and PRG inhibited insulin secretion in response to 16 mM glucose-stimulating concentrations. However, in the basal state (3 mM glucose), the insulin levels in response to DEX treatment were unchanged, and the three hormones together maintained higher insulin release. There were no changes of protein levels of GLUT2 or glucokinase (GK), but PRL or PRG treatment increased GK activity, whereas DEX had an inhibitory effect on GK activity. alpha-Ketoisocaproate (alpha-KIC)-stimulated insulin secretion was also reduced by DEX alone or combined with PRL and PRG, suggesting that DEX may inhibit distal steps in the insulin-exocytotic process. PRL treatment increased the concentration of intracellular cAMP in response to 16 mM glucose, suggesting a role for cAMP in potentiation of insulin secretion, whereas DEX alone or combined with PRL and PRG reduced cAMP levels by increasing phosphodiesterase (PDE) activity. These data provide evidence that PRL and to a lesser extent PRG, which increase in early pregnancy, enhance basal and glucose-stimulated insulin secretion in part by increasing GK activity and amplifying cAMP levels. Glucocorticoid, which increases throughout gestation, counteracts only glucose-stimulated insulin secretion under high glucose concentrations by dominantly inhibiting GK activity and increasing PDE activity to reduce cAMP levels. These adaptations in the beta-cell may play an important role in maintaining the basal hyperinsulinemia of pregnancy while limiting the capacity of PRL and PRG to promote glucose-stimulated insulin secretion during late gestation.


3D local structure around Zn in Kti11p as a representative Zn-(Cys)4 motif as obtained by MXAN.
Zinc is an important component of many proteins that play key roles in transcription, translation, and catalysis. Kti11p, DESR1, both belonging to a protein family characterized by a CSL zinc finger domain, and the co-catalytic zinc-protein PML containing a Zn(2+) binding domain called RING or C(3)HC(4) finger are all structurally determined by NMR although the zinc sites are silent to this spectroscopical method. The comparison of X-ray absorption near-edge spectroscopy (XANES) data for the three proteins demonstrates that fingerprints effect is a reliable method for a primary characterization of ligand species. Ab initio full MS calculations performed by MXAN are applied to obtain chemical and stereo structural information around the Zn ion in Kti11p. For the first time this high-spatial resolution technique confirms the formation of a stable Zn tetrahedral configuration with four sulfur ligands, and returns extremely accurate bond angle information between ligands.


Computational Design of Phosphatidylinositol 3-Kinase Inhibitors.
One of the most sought-after therapeutic targets for treating human cancers is the phosphoinositide 3-kinase; PI3k is an integral part of the PI3K/protein kinase B signaling arcade. This pathway is frequently activated in malignancies. Drug resistance and dose-limiting adverse effects are currently associated challenges with the existing anticancer chemotherapy. Therefore, in this research, a series of pyrimidine derivatives were designed and evaluated against human PI3K by using molecular docking analysis. The docking results were further verified by molecular dynamic simulation, which analyzed the strength of the macromolecular complex with respect to time. Compounds IV and XIV were found to be the most potent inhibitors of the human PI3K receptor with a high degree of stability within the active site of the target receptor for a timeframe of 50 ns. Thus, both of these compounds could be important drug candidates for the development of PI3K inhibitors as a prospective anticancer agent.


Association between cytokines in induced sputum and severity of chronic obstructive pulmonary disease.
Cytokines are known to be increased in induced sputum in chronic obstructive pulmonary disease (COPD). In this study, the relationship between the levels of interleukin-6 (IL-6), interleukin-8 (IL-8), and tumour necrosis factor-alpha (TNF-alpha) in induced sputum of patients with exacerbation of COPD, and the severity of the disease, pulmonary function tests (PFT), arterial blood gases (ABG) were studied. Twenty-four patients with exacerbation of COPD were included in the study. The patients were grouped according to their PFT into two as: Group 1 (FEV1 below 50% of the predicted value, severe-very severe COPD, n=12) and, Group 2 (FEV1 above 50% of the predicted value, mild-moderate COPD, n=12). The levels of IL-6, IL-8 and TNF-alpha in induced sputum of the subjects were measured. The mean levels of IL-6, IL-8 and TNF-alpha in induced sputum were found to be higher in Group 1 (severe-very severe COPD) than in Group 2 (mild-moderate COPD). The differences in IL-6 and IL-8 levels between groups were statistically significant (P<0.05). A significant correlation was observed between the IL-6 value and FEV(1) (r=-0.435, P=0.034), FEV1/FVC (r=-0.446, P=0.029), PaO2 (r=-0.711, P=0.000), SaO2 (r=-0.444, P=0.030) and disease duration (r=0.427, P=0.037), respectively. Also, the level of IL-8 in induced sputum was inversely correlated with FEV1 (r=-0.562, P=0.004), PaO2 (r=-0.540, P=0.006) and SaO2 (r=-0.435, P=0.034). However, all three cytokines were positively correlated with the smoking load (r=0.653, P=0.001; r=0.439, P=0.032; r=0.649, P=0.001). We conclude, therefore, that in exacerbated COPD cases with greater degrees of obstruction of the airways have higher levels of cytokines in induced sputum. This can be interpreted to mean that these cytokines are related to the clinical parameters like the ABG and PFT and seem to be the determinant of the severity of the disease.


5-methyltetrahydrofolate administration is associated with prolonged survival and reduced inflammation in ESRD patients.
Hemodialysis (HD) patients have a greatly increased risk of cardiovascular morbidity and mortality. For this reason, attempts are often made to normalize hyperhomocysteinemia. This randomized prospective study sought to determine which risk factors are predictors of mortality and whether high doses of folates or 5-methyltetrahydrofolate (5-MTHF) could improve hyperhomocysteinemia and survival in HD patients. 341 patients were divided into two groups: group A was treated with 50 mg i.v. 5-MTHF, and group B was treated with 5 mg/day oral folic acid. Both groups received i.v. vitamin B(6) and B(12). By dividing patients into C-reactive protein (CRP) quartiles, group A had the highest survival for CRP <12 mg/l, whereas no survival difference was found for group B. CRP was the only predictive risk factor for death (RR 1.17, range 1.04-1.30, p = 0.02). Dialysis age, hyperhomocysteinemia, methylenetetrahydrofolate reductase polymorphism, albumin, lipoprotein (a) and folate did not influence mortality risk. Survival in group A was higher than that in group B, namely 36.2 +/- 20.9 vs. 26.1 +/- 22.2 months (p = 0.003). Our results suggest that CRP, but not hyperhomocysteinemia, is the main risk factor for mortality in HD patients receiving vitamin supplements. Intravenous 5-MTHF seems to improve survival in HD patients independent from homocysteine lowering.


The Long-Term Course of Atopic Dermatitis.
Atopic dermatitis (AD) is a chronic, relapsing condition, meaning that the intensity of symptoms usually fluctuates over time. Changes in skin physiology may be evident from birth, suggesting that AD may be a lifelong condition marked by intermittent symptoms/disease activity. Methodological considerations for studying the long-term course of AD are reviewed in detail. Improved measurement of the frequency and duration of active disease periods can help to elucidate more about the clinical course AD and the role of treatment in long-term outcomes.


The delivery of dignity when caring for older people.
Everyone has a different understanding of what it means to have dignity, however, it is commonly associated with being in control and valued. The dignity of older people is therefore vulnerable as they are likely to be in positions of reduced control over their wellbeing. The six senses framework describes the characteristics of a health and social care environment which can promote dignity. The dignity of older people with dementia or near the end of life can be particularly at risk, however there are practical steps which can make a difference. Help the Aged has produced a series of guides to help medical practitioners and hospital staff care for vulnerable older people in the right way.


Microscopic description of elementary growth processes and classification of structural defects in pentacene thin films.
Elementary growth processes such as kink initiation, adding a molecule to a kink, and adding a molecule between two neighboring kinks and between two grains are theoretically studied in pentacene films by adding one molecule at a time to a predefined aggregate. For each molecule, the potential energy surface is calculated using the MM3 molecular mechanics force field, which allowed one to identify useful parameters like the energy barrier for diffusion and the energy to create kinks, as well as defect configurations. Depending on the properties of the potential energy surface and the resulting growth-condition-dependent probabilities of initiating defect configurations in the film, three types of pentacene defects are identified: a thermally activated defect, an intrinsic defect, and a kinetic defect. Upon film growth, most defects relax into the ideal crystal configuration. Bulk defects that resist relaxation have densities lower than 10(16) defects/cm3 at typical growth conditions. Grain boundary defects, on the other hand, are very stable. Moreover, interstitial molecules at grain boundaries are identified as a source of compressive stress.


Fourier Transform Microwave Spectrum of Propene-3-d1 (CH2═CHCH2D), Quadrupole Coupling Constants of Deuterium, and a Semiexperimental Equilibrium Structure of Propene.
The ground-state rotational spectrum of propene-3-d1, CH2═CHCH2D, was measured by Fourier transform microwave spectroscopy. Transitions were assigned for the two conformers, one with the D atom in the symmetry plane (S) and the other with the D atom out of the plane (A). The energy difference between the two conformers was calculated to be 6.5 cm-1, the S conformer having lower energy. The quadrupole hyperfine structure due to deuterium was resolved and analyzed for both conformers. The experimental quadrupole coupling and the centrifugal distortion constants compared favorably to their ab initio counterparts. Ground-state rotational constants for the S conformer are 40582.157(9), 9067.024(1), and 7766.0165(12) MHz. Ground-state rotational constants for the A conformer are 43403.75(3), 8658.961(2), and 7718.247(2) MHz. For the A conformer, a small tunneling splitting (19 MHz) due to internal rotation was observed and analyzed. Using the new rotational constants of this work as well as those previously determined for the 13C species and for some deuterium-substituted species from the literature, a new semiexperimental equilibrium structure was determined and its high accuracy was confirmed. The difficulty in obtaining accurate coordinates for the out-of-plane hydrogen atom is discussed.


Pulsatile portal vein insulin delivery enhances hepatic insulin action and signaling.
Insulin is secreted as discrete insulin secretory bursts at ~5-min intervals into the hepatic portal vein, these pulses being attenuated early in the development of type 1 and type 2 diabetes mellitus (T2DM). Intraportal insulin infusions (pulsatile, constant, or reproducing that in T2DM) indicated that the pattern of pulsatile insulin secretion delivered via the portal vein is important for hepatic insulin action and, therefore, presumably for hepatic insulin signaling. To test this, we examined hepatic insulin signaling in rat livers exposed to the same three patterns of portal vein insulin delivery by use of sequential liver biopsies in anesthetized rats. Intraportal delivery of insulin in a constant versus pulsatile pattern led to delayed and impaired activation of hepatic insulin receptor substrate (IRS)-1 and IRS-2 signaling, impaired activation of downstream insulin signaling effector molecules AKT and Foxo1, and decreased expression of glucokinase (Gck). We further established that hepatic Gck expression is decreased in the HIP rat model of T2DM, a defect that correlated with a progressive defect of pulsatile insulin secretion. We conclude that the physiological pulsatile pattern of insulin delivery is important in hepatic insulin signaling and glycemic control. Hepatic insulin resistance in diabetes is likely in part due to impaired pulsatile insulin secretion.


Lamprey fructose-1,6-bisphosphate aldolase: characterization of the muscle-type and non-muscle-type isozymes.
To study evolutionary aspects of fructose-1,6-bisphosphate (Fru-1,6-P2) aldolase during deuterostomian evolution, we have purified and characterized aldolases from the muscle and liver of lamprey (Entosphenus japonicus). Aldolase from the skeletal muscle and liver was identified to be the muscle-type isozyme and the non-muscle-type isozyme that was encoded by cDNAs M8 and L3, respectively, as described previously (Zhang, R., Yatsuki, H., Kusakabe, T., Iwabe, Miyata, T., Imai, T., Yoshida, M., and Hori, K., J. Biochem. 117, 545-553, 1995). The muscle-type isozyme has properties similar to vertebrate aldolase A, while the non-muscle-type isozyme shows a similarity to bacterial class I aldolase and vertebrate aldolase C but not to aldolase B, the liver-type aldolase, in terms of kinetic parameters: the Kcat values toward Fru-1,6-P2 and Fru-1-P, the Fru-1,6-P2/Fru-1-P activity ratio, and the Km values toward these substrates. The two enzymes have tetrameric forms with a molecular mass of approximately 160,000 and have similar pH optimum. The muscle-type and non-muscle-type isozymes from the tissues show different electrophoretic mobility; the muscle-type isozyme moves much faster than the non-muscle-type isozyme toward anodic side. The recombinant muscle-type and non-muscle-type aldolases gave similar characteristics as those from the tissues. The results presented in this paper, together with the data presented in the previous paper, strongly suggest that in lamprey it is possible to have two types of aldolase isozymes rather than one or three isozymes.


VraR Binding to the Promoter Region of agr Inhibits Its Function in Vancomycin-Intermediate Staphylococcus aureus (VISA) and Heterogeneous VISA.
Acquisition of vancomycin resistance in Staphylococcus aureus is often accompanied by a reduction in virulence, but the mechanisms underlying this change remain unclear. The present study was undertaken to investigate this process in a clinical heterogeneous vancomycin-intermediate S. aureus (hVISA) strain, 10827; an hVISA reference strain, Mu3; and a VISA reference strain, Mu50, along with their respective series of vancomycin-induced resistant strains. In these strains, increasing MICs of vancomycin were associated with increased expression of the vancomycin resistance-associated regulator gene (vraR) and decreased expression of virulence genes (hla, hlb, and coa) and virulence-regulated genes (RNAIII, agrA, and saeR). These results suggested that VraR might have a direct or indirect effect on virulence in S. aureus In electrophoretic mobility shift assays, VraR did not bind to promoter sequences of hla, hlb, and coa genes, but it did bind to the agr promoter region. In DNase I footprinting assays, VraR protected a 15-nucleotide (nt) sequence in the intergenic region between the agr P2 and P3 promoters. These results indicated that when S. aureus is subject to induction by vancomycin, expression of vraR is upregulated, and VraR binding inhibits the function of the Agr quorum-sensing system, causing reductions in the virulence of VISA/hVISA strains. Our results suggested that VraR in S. aureus is involved not only in the regulation of vancomycin resistance but also in the regulation of virulence.


Influence of p53 status on radiation and 5-flourouracil synergy in pancreatic cancer cells.
While p53 protein plays an important role in the regulation of radiosensitivity and chemosensitivity in many tumors, the role of p53 in the combined management of tumors that harbor mutations in the p53 gene have not been fully defined. This study was undertaken to evaluate the impact of wild-type or mutant p53 status on the synergistic effects of 5-Fluorouracil (5-FU) and radiation (XRT) in pancreatic tumors. Three pancreatic tumor cell lines, one containing wild-type functional p53 (Capan-2) and two containing mutant p53 (Panc-1 and MIA PaCa-2), were used in this study. Radiation-induced p53 and p21(waf1/cip1) protein expression was determined by Western blot analysis. Radiation induced Thymidylate Synthase (TS) mRNA expression was determined by 32P-RT-PCR. The effect of 5-FU, radiation, and radiation +5-FU on the growth and colony-forming ability of Capan-2, Panc-1 and MIA PaCa-2 was determined by clonogenic assays respectively. Radiation elevated p53 and p21(waf1/cip1) levels in Capan-2 cells. No elevation of p53 and p21(waf1/cip1) was evident in Panc-1. MIA PaCa-2 cells showed down-regulation of p21(waf1/cip1) with no elevation of p53 protein. Clonogenic assays showed enhanced radiosensitizing effect when 5-Fluorouracil was added to cell lines lacking functional p53. In wild-type p53 Capan-2 cells, radiation up-regulated TS mRNA levels. High basal levels of TS mRNA were detected in p53 mutant cell lines with no evident induction by radiation. Our results confirm that p53 status has a significant impact on radiation sensitivity with wild-type p53 cells being significantly more radiosensitive than mutant cell lines. When XRT and 5-FU were combined, this led only to an additive effect in wild-type cell lines and a synergistic effect in mutant cell lines.


Fungal infection involvement in primary biliary cirrhosis: A review of 2 cases.
The present study aimed to analyze the imaging, clinical and pathological features of fungal infection involvement in primary biliary cirrhosis (PBC) by retrospectively analyzing and reviewing the features of two patients with fungal infection involvement in PBC. Both patients were female. One patient had a confirmed diagnosis of PBC. The other patient had confirmed Sjogren syndrome and PBC. The two cases of PBC were infected with fungal infection after treatment with hormonal and immunosuppressive agents. RCR of sputum confirmed Pneumocystis spp. infection in the patient with PBC alone. The mucormycosis infection was confirmed in the other patient after pathological examination of a renal biopsy. The state of the illnesses progressed quickly and both patients ultimately succumbed to their conditions. The patient prognosis of fungal infection involvement PBC is poor. Patients treated with long-term hormone and immunosuppressive agents should be monitored.


Photothermal deflection in multilayer coatings: modeling and experiment.
A model of the photothermal deflection signal in multilayer coatings is presented that takes into account optical interference effects and heat flow within the stack. Measurements are then taken of high-reflectivity HfO2/SiO2 ultraviolet mirrors made by plasma ion assisted deposition and compared to calculations. Good agreement is found between the experimental results and the model. Using this model for the calibration and the setup described, one can measure absorption in multilayer coatings accurately down to 10(-7) of the incident power.


[Aging and anti-aging of oocytes].
The functions of organs decrease as the age increases. The fecundity of women also decreases due to mainly the decreasing quality of oocytes. The recent change of life style makes the age of infertility patients elder and infertility treatments more difficult. The therapeutic strategy for the elder infertility patients is still chaotic. The precise evaluation for the ageing in the female genital organs should be developed and the treatment for the failure of fecundity due to aging must be overcome.


Development and validation of risk scoring system for prediction of cancer cervix.
A Hospital based group matched case-control study was conducted to devise a risk scoring system for the prediction of cancer cervix at the Gynecology Clinic, Government Medical College Hospital, Nagpur, India. The study consisted of 230 cases of cancer cervix (histopathologically confirmed) and equal number of controls, group matched for age. The risk factors considered were Illiteracy, long duration of married life (>25 years), Early Menarche (<13 years), marital status (widow, separated, divorcee), multiparity (> 3), h/o abortion, h/o tobacco use, h/o passive smoking, poor genital hygiene, (grade III & IV) and low socioeconomic status. Statistical Analysis included unconditional multiple logistic regression analysis Receiver Operating Characteristic (ROC) curve analysis. The overall predictive accuracy was calculated by Wilcoxon statistic as an equivalent of area under ROC curve. Five risk factors, illiteracy, poor genital hygiene, long duration of married life, multiparity and early menarche were identified to be significantly associated with cancer cervix. These factors were given statistical weights of 13, 10, 7, 5 and 5 respectively. A total score of 21 was found to be the best cut off for prediction and the overall predictive accuracy of the risk scoring system was calculated to be 0.74(0.67 - 0.81). In case of consistent further validation using other data sets this additive risk scoring system can be used for reducing the cost of universal screening by subjecting only high-risk subjects to laboratory screening procedure (Pap smear) in population setting.


Mercury biomarkers and DNA methylation among Michigan dental professionals.
Modification of the epigenome may be a mechanism underlying toxicity and disease following chemical exposure. Animal and human data suggest that mercury (Hg) impacts DNA methylation. We hypothesize that methylmercury and inorganic Hg exposures from fish consumption and dental amalgams, respectively, may be associated with altered DNA methylation at global repetitive elements (long interspersed elements, LINE-1) and candidate genes related to epigenetic processes (DNMT1) and protection against Hg toxicity (SEPW1, SEPP1). Dental professionals were recruited at Michigan Dental Association (MDA) meetings in 2009 and 2010. Subjects (n=131) provided survey data (e.g. exposure sources, demographics) and biological samples for Hg measurement and epigenetic analysis. Total Hg was quantified via atomic absorption spectrophotometry in hair and urine, indicative of methylmercury and inorganic Hg exposures, respectively. Global repetitive and candidate gene methylation was quantified via pyrosequencing of bisulfite converted DNA isolated from buccal mucosa. Hair Hg (geometric mean (95% CI): 0.37 (0.31-0.44) µg/g) and urine Hg (0.70 (0.60-0.83) µg/L) were associated with sources of exposure (fish consumption and dental amalgams, respectively). Multivariable linear regression revealed a trend of SEPP1 hypomethylation with increasing hair Hg levels, and this was significant (P<0.05) among males. The trend remained when excluding non-dentists. No significant relationships between urine Hg and DNA methylation were observed. Thus, in a limited cohort, we identified an association between methylmercury exposure and hypomethylation of a potentially labile region of the genome (SEPP1 promoter), and this relationship was gender specific.


Predator identity more than predator richness structures aquatic microbial assemblages in Sarracenia purpurea leaves.
The importance of predators in influencing community structure is a well-studied area of ecology. However, few studies test ecological hypotheses of predation in multi-predator microbial communities. The phytotelmic community found within the water-filled leaves of the pitcher plant, Sarracenia purpurea, exhibits a simple trophic structure that includes multiple protozoan predators and microbial prey. Using this system, we sought to determine whether different predators target distinct microorganisms, how interactions among protozoans affect resource (microorganism) use, and how predator diversity affects prey community diversity. In particular, we endeavored to determine if protozoa followed known ecological patterns such as keystone predation or generalist predation. For these experiments, replicate inquiline microbial communities were maintained for seven days with five protozoan species. Microbial community structure was determined by 16S rRNA gene amplicon sequencing (iTag) and analysis. Compared to the control (no protozoa), two ciliates followed patterns of keystone predation by increasing microbial evenness. In pairwise competition treatments with a generalist flagellate, prey communities resembled the microbial communities of the respective keystone predator in monoculture. The relative abundance of the most common bacterial Operational Taxonomic Unit (OTU) in our system decreased compared to the control in the presence of these ciliates. This OTU was 98% similar to a known chitin degrader and nitrate reducer, important functions for the microbial community and the plant host. Collectively, the data demonstrated that predator identity had a greater effect on prey diversity and composition than overall predator diversity.


Vibrio cholerae VciB Mediates Iron Reduction.
Vibrio cholerae is the causative agent of the severe diarrheal disease cholera. V. cholerae thrives within the human host, where it replicates to high numbers, but it also persists within the aquatic environments of ocean and brackish water. To survive within these nutritionally diverse environments, V. cholerae must encode the necessary tools to acquire the essential nutrient iron in all forms it may encounter. A prior study of systems involved in iron transport in V. cholerae revealed the existence of vciB, which, while unable to directly transport iron, stimulates the transport of iron through ferrous (Fe2+) iron transport systems. We demonstrate here a role for VciB in V. cholerae in which VciB stimulates the reduction of Fe3+ to Fe2+, which can be subsequently transported into the cell with the ferrous iron transporter Feo. Iron reduction is independent of functional iron transport but is associated with the electron transport chain. Comparative analysis of VciB orthologs suggests a similar role for other proteins in the VciB family. Our data indicate that VciB is a dimer located in the inner membrane with three transmembrane segments and a large periplasmic loop. Directed mutagenesis of the protein reveals two highly conserved histidine residues required for function. Taken together, our results support a model whereby VciB reduces ferric iron using energy from the electron transport chain.IMPORTANCEVibrio cholerae is a prolific human pathogen and environmental organism. The acquisition of essential nutrients such as iron is critical for replication, and V. cholerae encodes a number of mechanisms to use iron from diverse environments. Here, we describe the V. cholerae protein VciB that increases the reduction of oxidized ferric iron (Fe3+) to the ferrous form (Fe2+), thus promoting iron acquisition through ferrous iron transporters. Analysis of VciB orthologs in Burkholderia and Aeromonas spp. suggest that they have a similar activity, allowing a functional assignment for this previously uncharacterized protein family. This study builds upon our understanding of proteins known to mediate iron reduction in bacteria.


Deep Learning in Medical Image Analysis.
This review covers computer-assisted analysis of images in the field of medical imaging. Recent advances in machine learning, especially with regard to deep learning, are helping to identify, classify, and quantify patterns in medical images. At the core of these advances is the ability to exploit hierarchical feature representations learned solely from data, instead of features designed by hand according to domain-specific knowledge. Deep learning is rapidly becoming the state of the art, leading to enhanced performance in various medical applications. We introduce the fundamentals of deep learning methods and review their successes in image registration, detection of anatomical and cellular structures, tissue segmentation, computer-aided disease diagnosis and prognosis, and so on. We conclude by discussing research issues and suggesting future directions for further improvement.


Predictors for delayed ventriculoperitoneal shunt placement after external ventricular drain removal in patients with subarachnoid hemorrhage.
Hydrocephalus after subarachnoid hemorrhage (SAH) requires temporary cerebrospinal fluid (CSF) drainage using an external ventricular drain (EVD). This drain is removed if patients pass a clamp trial, or a ventriculoperitoneal shunt (VPS) is placed. Little is known about the risk factors for delayed VPS placement in patients who pass a clamp trial and have their EVD removed. In order to explore the risk factors associated with delayed VPS placement, we studied a retrospective cohort of SAH patients at our institution. We performed a retrospective analysis of SAH patients who had an EVD placed between January 2008 and June 2012 at our institution. We extracted demographic, imaging, and CSF data from the medical record and analyzed risk factors associated with delayed VPS placement. Of 91 patients who passed a clamp trial and had their EVD removed, 12 (13%) required delayed VPS placement at a median of 54 (interquartile range: 15-75) days after EVD removal. After multivariate analysis, risk factors for delayed VPS placement included increased CSF protein concentration within the first 7 days of EVD placement (OR: 1.02, CI: 1-1.04, p=0.023) and increased third ventricular diameter prior to EVD removal (OR: 1.59, CI: 1.11-2.6, p=0.026). Patients with increased CSF protein concentration at time of EVD placement and those with increased third ventricular diameter at time of EVD removal should be carefully monitored for development of delayed hydrocephalus.


The odontogenic keratocyst: orthokeratinized variant.
The histopathologic and clinical features of sixty cases of orthokeratinized odontogenic cysts were compared with those of odontogenic keratocysts (typically parakeratinized). According to the results of this study, the orthokeratinized odontogenic cyst appears to be a distinct clinicopathologic entity. This cyst is histologically characterized by a thin, uniform, epithelial lining with orthokeratinization and a subjacent granular cell layer. The basal cells are usually cuboidal or flattened. Clinically, the orthokeratinized cyst is a single cyst, shows a predilection for males, and is most often found in the second to the fifth decade, it appears most commonly as a dentigerous cyst in the posterior mandible. The orthokeratinized cyst shows little clinical aggressiveness. Follow-up of twenty-four patients revealed only one recurrence; of nineteen patients evaluated, none had features of the basal cell nevus-bifid rib syndrome. It is suggested that this cyst be called odontogenic keratocyst, orthokeratinized variant.


Increase in lifetime adverse drug reactions, service utilization, and disease severity among patients who will start COX-2 specific inhibitors: quantitative assessment of channeling bias and confounding by indication in 6689 patients with rheumatoid arthritis and osteoarthritis.
Nonrandom assignment of therapy in observational studies and clinical practice can be accompanied by channeling bias and confounding by indication. This in turn can lead to unreliable conclusions about treatment effectiveness. Although widely acknowledged as important, no studies in rheumatology have measured the extent of these biases. We identified variables contributing to confounding and investigated the strength of the confounding effect. Analytical methods (propensity scores) are available to mitigate the effect of nonrandom assignment if the full extent of confounding can be understood. A population of 6637 patients with rheumatoid arthritis (RA) and osteoarthritis (OA) from the practices of 433 US rheumatologists completed 2 sets of detailed questionnaires concerning (1) the last 6 months in 1998 and (2) the first 6 months of 1999, generally prior to and after the release of celecoxib and rofecoxib. Patients who received the COX-2 specific inhibitors in period 2 were identified (n = 1517), and their characteristics were compared to the 5120 who did not start a new COX-2 specific inhibitor during Period 1. Patients starting a new COX-2 specific inhibitor had a greater lifetime history of adverse reactions of all kinds, but particularly gastrointestinal adverse drug reactions. They also had more severe scores for pain, functional disability, fatigue, helplessness, and global severity, and they used more inpatient and outpatients services than patients who would not switch to COX-2 specific inhibitors. Confounding by indication and channeling bias result in an overall increase in severity of about 25% for the above measures. Observational studies should account for these biases by a broadly defined propensity score that includes the variables identified in this report. These observations are germane to observational studies of disease modifying antirheumatic drugs and biologics, as well, and suggest the need for careful control of confounders when assessing treatment effects in rheumatic disease observational studies.


The relationship between maternal-fetal attachment and maternal self-efficacy in Iranian women: a prospective study.
This prospective study was conducted to determine the relationship between maternal-fetal attachment and maternal self-efficacy. Maternal self-efficacy contributes significantly to the mental health of the mother and infant. Maternal-fetal attachment facilitates maternal role attainment and might improve maternal self-efficacy. This study was conducted on 242 women. The data collection tools used included a socio-demographic and obstetric questionnaire, Cranley's Maternal-Fetal Attachment Scale, the Edinburgh Postnatal Depression Scale and the Maternal Self-Efficacy Scale. A multivariate linear regression was used to estimate the extent to which maternal-fetal attachment affects maternal self-efficacy. Pearson's correlation test showed a moderate but significant positive correlation between maternal-fetal attachment and self-efficacy (r = 0.48, P < 0.001). The highest correlation with self-efficacy was observed in the domain of 'differentiation of self from fetus' (r = 0.43) and the lowest in the domain of 'giving of self' (r = 0.25). According to the multivariate linear regression, the domain of 'Interaction with fetus' had a significant relationship with maternal self-efficacy (P = 0.009). Maternal-fetal attachment would appear to be a factor related to postpartum maternal self-efficacy which suggests identifying and supporting women with low self-efficacy may increase maternal-fetal attachment and thereby maternal self-efficacy.


Rational design of a high-affinity, fast, red calcium indicator R-CaMP2.
Fluorescent Ca(2+) reporters are widely used as readouts of neuronal activities. Here we designed R-CaMP2, a high-affinity red genetically encoded calcium indicator (GECI) with a Hill coefficient near 1. Use of the calmodulin-binding sequence of CaMKK-α and CaMKK-β in lieu of an M13 sequence resulted in threefold faster rise and decay times of Ca(2+) transients than R-CaMP1.07. These features allowed resolving single action potentials (APs) and recording fast AP trains up to 20-40 Hz in cortical slices. Somatic and synaptic activities of a cortical neuronal ensemble in vivo were imaged with similar efficacy as with previously reported sensitive green GECIs. Combining green and red GECIs, we successfully achieved dual-color monitoring of neuronal activities of distinct cell types, both in the mouse cortex and in freely moving Caenorhabditis elegans. Dual imaging using R-CaMP2 and green GECIs provides a powerful means to interrogate orthogonal and hierarchical neuronal ensembles in vivo.


Primary Malignancy Is an Independent Determinant of Morbidity and Mortality after Liver Resection.
Although outcomes after liver resection have improved, there remains considerable perioperative morbidity and mortality with these procedures. Studies suggest a primary liver cancer diagnosis is associated with poorer outcomes, but the extent to which this is attributable to a higher degree of hepatic dysfunction is unclear. To better delineate this, we performed a matched pair analysis of primary versus metastatic malignancies using a national database. The American College of Surgeons National Surgical Quality Improvement Program (2005-2013) was analyzed to select elective liver resections. Diagnoses were sorted as follows: 1) primary liver cancers and 2) metastatic neoplasms. A literature review identified factors known to impact hepatectomy outcomes; these variables were evaluated by a univariate analysis. The most predictive factors were used to create similar groups from each diagnosis category via propensity matching. Multivariate regression was used to validate results in the wider study population. Outcomes were compared using chi-squared test and Fisher exact test. Matched groups of 4838 patients were similar by all variables, including indicators of liver function. A number of major complications were significantly more prevalent with a primary diagnosis; overall major morbidity rates in the metastatic and primary groups were 29.3 versus 41.6 per cent, respectively. The mortality rate for primary neoplasms was 4.6 per cent (vs 1.6%); this represents a risk of death nearly three-times greater (95% confidence interval = 2.20-3.81, P < 0.0001) in cancers of hepatic origin. Hepatectomy carries substantially higher perioperative risk when performed for primary liver cancers, independent of hepatic function and resection extent. This knowledge will help to improve treatment planning, patient education, and resource allocation in oncologic liver resection.


KNUCKLES (KNU) encodes a C2H2 zinc-finger protein that regulates development of basal pattern elements of the Arabidopsis gynoecium.
Flowers of the parthenocarpic knuckles mutant are conditionally male sterile and contain ectopic stamens and carpels that originate from placental tissue within developing gynoecia. The mutation was mapped to a 123 Kb interval on chromosome 5 using molecular markers. All aspects of the knuckles phenotype could be complemented by a genomic fragment from the region which contained the annotated MAC12.2 gene. A guanine to adenine transition within a predicted C2H2 zinc finger-encoding region of MAC12.2 causes the second of two critical zinc-binding cysteine residues to be replaced by a tyrosine. Transgenic plants in which translational fusions of the GUS reporter to KNUCKLES were driven by the presumptive KNUCKLES promoter indicate that the gene is expressed first in developing carpel primordia, and later in stamens and ovules of flower buds. In situ hybridization experiments showed a broader pattern of transcript localization, suggesting that post-transcriptional regulatory mechanisms may limit KNUCKLES protein accumulation and localization. Based on genetic evidence and the presence of a carboxy-terminal motif demonstrated by others to function as an active repression domain, we propose that KNUCKLES might function as a transcriptional repressor of cellular proliferation that regulates floral determinacy and relative size of basal pattern elements along the proximo-distal axis of the developing Arabidopsis gynoecium.


Use of a paramagnetic substance, colloidal manganese sulfide, as an NMR contrast material in rats.
Paramagnetic pharmaceuticals ( magnetopharmaceuticals ) that are suitably distributed into specific organ systems or diseased sites might be clinically useful for tissue contrast enhancement in nuclear magnetic resonance images. To determine whether an insoluble magnetopharmaceutical might be useful in such service, we investigated the effect of a colloidal preparation of manganese sulfide ( MnSC ) upon liver and lung spin-lattice relaxation times (T1) in rats following intravenous administration. NMR tissue sample measurements were made at 24 MHz, ahd showed that after MnSC treatment, liver T1 values--and to a lesser extent lung T1 values--were depressed below control values. Liver manganese content (as determined by flame atomic absorption spectrophotometry) increased in proportion to the dose of MnSC , and the reciprocal of the liver T1 values also increased in proportion to the dose of MnSC .


On the analyst's neutrality.
It is by the application of the principle of neutrality, born of his respect for the essential otherness of the patient, that the analyst focuses the dyadic analytic work in the service of the patient's growing self-analytic capacity. Thus, the general principle of neutrality is distinguished from the technical tactic of abstinence, the latter being a specific function utilized to facilitate and foster analytic regression. Neutrality can be defined as it applies to the major subfunctions of the analyst's work ego. Perception of the patient's intrapsychic processes (both empathically and cognitively) requires a neutrality of appearance on the analyst's part in order to minimize the distortion of the unfolding transference neurosis. Integration and understanding of the patient's communications require mastery and neutralization of the analyst's own internal processes in order to minimize countertransferential distortions. Appropriate interpretive intervention requires neutrality of action, i.e., mastery of impulses related to power, neutralizing them into the service of the analytic work; tact is defined as a specific psychoanalytic function in this regard. Collaborative ignorance is examined as a specific instance of false neutrality. In this an analytic guise serves to mask a countertransferential conflict. Neutrality serves as an overriding technical principle, not an imperative for perfectionism . Factors intrinsic to the analytic process also influence the application of this principle.


Hematologic and immunologic responses of Holstein calves to a fire ant toxicant.
Holstein calves (3 to 5 months of age) were used to develop an animal model sensitive to environmental toxicants. In the present study, the fire ant toxicant AMDRO was fed (113.5 g/day/calf) to weanling castrated calves (9 test and 9 controls) for 7 weeks. As early as 14 days after the start of the AMDRO feeding, leukopenia was observed. Differential counts revealed significant non-transient decreases in lymphocytes and eosinophils. Eosinopenia was observed from days 21 to 49 of AMDRO treatment. Variability in hematocrit and hemoglobin values in treated and control calves precluded making a determination of trends due to toxicant exposure. The AMDRO treatment did not produce significant decreases in primary or secondary antibody responses to keyhole limpet hemocyanin or to Brucella abortus vaccination. It also did not produce suppression of cellular immunity, as determined by delayed-type hypersensitivity response to bacille Calmette-Guérin. Potential effects of AMDRO on thermoregulatory mechanisms were indicated by significant depressions of rectal temperature observed after 25, 32, and 39 days' treatment.


Effects of carbon dioxide insufflation on anastomosis remodeling at a carotid artery site in rabbits.
Use of carbon dioxide (CO2) insufflation (CDI) on the surgical field during heart surgery has become widespread, and in some units routine. To assess the effects of CDI on endothelial dysfunction in a carotid artery model in rabbits. Twelve randomly selected rabbits were divided into two groups. Right carotid arteries of the animals were transected and sutured with running suture technique. Then, 1 l/min CO2 insufflation was initiated with a 45° angle. In the control group, the anastomotic field was irrigated with 0.1 ml/s flow of 0.9% saline. At day 28, the carotid artery segments were removed and prepared for histological specimens. In the cross-sections of the control group vessel samples, thickening of the tunica intima was observed. Scoring the quantity of endothelial nitric oxide synthase (e-NOS) and α-smooth muscle actin (α-SMA) positive staining revealed a nonsignificant difference between the experimental groups (p = 0.07). In the CO2 group, the intimal hyperplasia (p = 0.2) and the thickness of the tunica media (p = 0.2) were indistinguishable when compared to the control group. The mean luminal diameters and luminal areas of the experimental groups were all evaluated by histomorphometry and a significant differences was found between luminal areas (p = 0.016). On the other hand, no significant difference was found between mean luminal diameters (p = 0.055). Our study indicated that CDI can affect endothelial cell damage and the mean luminal diameters.


The impact of drying techniques on phenolic compound, total phenolic content and antioxidant capacity of oat flour tarhana.
In this study, the changes in phenolic composition, total phenolic content, and antioxidant capacity of tarhanas supplemented with oat flour (OF) at the levels of 20-100% (w/w) after three drying treatments (sun-, oven-, and microwave drying) were investigated. A total of seventeen phenolic standards have been screened in tarhanas, and the most abundant flavonol and phenolic acid compounds were kaempferol (23.62mg/g) and 3-hydroxy-4-metoxy cinnamic acid (9.60mg/g). The total phenolic content amount gradually increased with the addition of OF to tarhana, but decidedly higher total phenolic content was found in samples oven dried at 55°C as compared with other methods. The microwave- and oven dried tarhana samples showed higher TEACDPPH and TEACABTS values than those dried with the other methods, respectively, in higher OF amounts. Consequently, oven- and microwave-drying can be recommended to retain the highest for phenolic compounds as well as maximal antioxidant capacity in OF supplemented tarhana samples.


Optical power detector with broad spectral coverage, high detectivity, and large dynamic range.
Optical power measurements are needed in practically all technologies based on light. Here, we report a general-purpose optical power detector based on the photoacoustic effect. Optical power incident on the detector's black absorber produces an acoustic signal, which is further converted into an electrical signal using a silicon-cantilever pressure transducer. We demonstrate an exceptionally large spectral coverage from ultraviolet to far infrared, with the possibility for further extension to the terahertz region. The linear dynamic range of the detector reaches 80 dB, ranging from a noise-equivalent power of 6 n W/H z to 600 mW (independent of signal averaging time).


GENETIC STRUCTURE OF GIANT CLAM (TRIDACNA MAXIMA) POPULATIONS IN THE WEST PACIFIC IS NOT CONSISTENT WITH DISPERSAL BY PRESENT-DAY OCEAN CURRENTS.
The Pacific marine biota, particularly species with long planktonic larval stages, are thought to disperse widely throughout the Pacific via ocean currents. The little genetic data available to date has supported this view in that little or no significant regional differentiation of populations has been found over large geographical distances. However, recent data from giant clams has demonstrated not only significant regional differentiation of populations, but routes of gene flow that run perpendicular to the main present-day ocean currents. Extensive surveys of genetic variation at eight polymorphic loci in 19 populations of the giant clam Tridacna maxima, sampled throughout the West and Central Pacific, confirmed that the patterns of variation seen so far in T. gigas were not unique to that species, and may reflect a fundamental genetic structuring of shallow-water marine taxa. Populations of T. maxima within highly connected reef systems like the Great Barrier Reef were panmictic (average FST < 0.003), but highly significant genetic differences between reef groups on different archipelagos (average FST = 0.084) and between West and Central Pacific regions (average FST = 0.156) were found. Inferred gene flow was high (Ne m usually > 5) between the Philippines and the Great Barrier Reef, between the Philippines and Melanesia (the Solomon Islands and Fiji), and between the Philippines and the Central Pacific island groups (Marshall Islands, Kiribati, Tuvalu and Cook Islands). Gene flow was low between these three sets of island chains (Ne m < 2). These routes of gene flow are perpendicular to present-day ocean currents. It is suggested that the spatial patterns of gene frequencies reflect past episodes of dispersal at times of lower sea levels which have not been erased by subsequent dispersal by present-day circulation. The patterns are consistent with extensive dispersal of marine species in the Pacific, and with traditional views of dispersal from the Indo-Malay region. However, they demonstrate that dispersal along present-day ocean surface currents cannot be assumed, that other mechanisms may operate today or that major dispersal events are intermittent (perhaps separated by several thousands of years), and that the nature and timing of dispersal of Pacific marine species is more complex than has been thought.


Microtubule dynamics and the neurodegenerative triad of Alzheimer's disease: The hidden connection.
Alzheimer's disease (AD) is the most common neurodegenerative disorder and is, on a histopathological level, characterized by the presence of extracellular amyloid plaques composed of the protein fragment Aβ, and intracellular neurofibrillary tangles, which contain the microtubule-associated protein tau in a hyperphosphorylated state. In AD defects in microtubule (MT) assembly and organization have also been reported; however, it is unclear whether MT abnormalities have a causal and early role in the disease process or represent a common end point downstream of the neurodegenerative cascade. Recent evidence indicates that microtubule-stabilizing drugs prevent axonopathy in animal models of tauopathies and reverse Aβ-induced loss of synaptic connectivity in an ex vivo model of amyloidosis. This could suggest that MT dysfunction connects some of the degenerative events and provides a useful target to simultaneously prevent several neurodegenerative processes in AD. Here, we describe how changes in the structure and dynamics of MTs are involved in the different aspects of the neurodegenerative triad of AD. We discuss evidence that MTs are affected both by tau-dependent and tau-independent mechanisms but appear to be regulated in a distinct way in different neuronal compartments. We argue that modulation of MT dynamics could be of potential benefit but needs to be precisely controlled in a cell and compartment-specific manner to avoid harmful side effects. This article is part of the series "Beyond Amyloid".


Relationship of tumor immunogenicity to concentration of the oncogen.
Tumors were induced sc in (BALB/c x DBA/2)F1 female mice by various concentrations of 3-methylcholanthrene (MCA) in paraffin pellets. There was an inverse relationship between MCA concentration and tumor latency (interval between MCA implantation and detection of gross tumor). The tumors were transplanted into syngeneic recipients, and material from this first transplant generation was used to immunize a series of syngeneic mice; any resulting growth was excised. Nonimmunized mice were controls. Immunized and control mice were irradiated and given an sc inoculation of a near-threshold number of tumor cells. Tumor growth from that inoculation was measured weekly in both groups and the antigenicity ratio (mean tumor size in controls/mean tumor size in immunized mice) was calculated. In a series of tumors with similar latencies, the only ones with high antigenicity ratios were those resulting from the high MCA concentration. The results suggested that tumors induced by low levels of oncogen may be good models of spontaneous neoplasia, strengthened the hypothesis that "spontaneous' tumors may actually result from low levels of oncogen, and indicated that neoplastic transformation and the development of immunogenicity are, at least in chemically induced tumors, independent changes that may be produced in the same cell when the concentration of oncogen is sufficient.


Determination of Anti-Adeno-Associated Viral Vector Neutralizing Antibodies in Patients With Heart Failure in the Cardiovascular Foundation of Colombia (ANVIAS): Study Protocol.
Recent progress in the pathophysiology of heart failure (HF) has led to the development of new therapeutic options such as gene therapy and the use of adeno-associated viral (AAV) vectors. Despite the promising results in early clinical trials of gene therapy for HF, various obstacles have been faced, such as the presence of neutralizing antibodies (NAbs) against the capsid vectors. NAb activity limits vector transduction levels and therefore diminishes the final therapeutic response. Recent studies evaluating the prevalence of NAbs in various populations found considerable geographic variability for each AAV serotype. However, the levels of NAbs in Latin American populations are unknown, becoming a limiting factor to conducting AAV vector therapeutic trials in this population. The goal of this study is to determine for the first time, the prevalence of anti-AAV NAbs for the serotypes 1, 2, and 9 in HF patients from the city of Bucaramanga, Colombia, using the in vitro transduction inhibition assay. We will conduct a cross-sectional study with patients who periodically attend the HF clinic of the Cardiovascular Foundation of Colombia and healthy volunteers matched for age and sex. For all participants, we will evaluate the NAb levels against serotypes AAV1, AAV2, and AAV9. We will determine NAb levels using the in vitro transduction inhibition assay. In addition, participants will answer a survey to evaluate their epidemiological and socioeconomic variables. Participation in the study will be voluntary and all participants will sign an informed consent document before any intervention. The project is in the first phase: elaboration of case report forms and the informed consent form, and design of the recruitment strategy. Patient recruitment is expected to begin in the spring of 2016. We expect to have preliminary results, including the titer of the viral vectors, multiplicity of infections that we will use for each serotype, and the general validation of the assay, at the end of 2016. The final results are expected mid-2017. This project is the first effort to evaluate NAb levels against AAV1, AAV2, and AAV9 serotypes in patients with HF in Latin America. Our results will allow us to check the cross-reactivity response between the serotypes assessed, to describe the epidemiological characteristics of the participant population, and to set up a link with earlier reports of NAb prevalence in the literature.


Microbial inactivation and cytotoxicity evaluation of UV irradiated coconut water in a novel continuous flow spiral reactor.
A continuous-flow UV reactor operating at 254nm wave-length was used to investigate inactivation of microorganisms including bacteriophage in coconut water, a highly opaque liquid food. UV-C inactivation kinetics of two surrogate viruses (MS2, T1UV) and three bacteria (E. coli ATCC 25922, Salmonella Typhimurium ATCC 13311, Listeria monocytogenes ATCC 19115) in buffer and coconut water were investigated (D10 values ranging from 2.82 to 4.54mJ·cm-2). A series of known UV-C doses were delivered to the samples. Inactivation levels of all organisms were linearly proportional to UV-C dose (r2>0.97). At the highest dose of 30mJ·cm-2, the three pathogenic organisms were inactivated by >5 log10 (p<0.05). Results clearly demonstrated that UV-C irradiation effectively inactivated bacteriophage and pathogenic microbes in coconut water. The inactivation kinetics of microorganisms were best described by log linear model with a low root mean square error (RMSE) and high coefficient of determination (r2>0.97). Models for predicting log reduction as a function of UV-C irradiation dose were found to be significant (p<0.05) with low RMSE and high r2. The irradiated coconut water showed no cytotoxic effects on normal human intestinal cells and normal mouse liver cells. Overall, these results indicated that UV-C treatment did not generate cytotoxic compounds in the coconut water. This study clearly demonstrated that high levels of inactivation of pathogens can be achieved in coconut water, and suggested potential method for UV-C treatment of other liquid foods. This research paper provides scientific evidence of the potential benefits of UV-C irradiation in inactivating bacterial and viral surrogates at commercially relevant doses of 0-120mJ·cm-2. The irradiated coconut water showed no cytotoxic effects on normal intestinal and healthy mice liver cells. UV-C irradiation is an attractive food preservation technology and offers opportunities for horticultural and food processing industries to meet the growing demand from consumers for healthier and safe food products. This study would provide technical support for commercialization of UV-C treatment of beverages.


Does Timing of Ventricular Tachycardia Ablation Affect Prognosis in Patients With an Implantable Cardioverter Defibrillator? Results From the Multicenter Randomized PARTITA Trial.
Optimal timing for catheter ablation of ventricular tachycardia is an important unresolved issue. There are no randomized trials evaluating the benefit of ablation after the first implantable cardioverter defibrillator (ICD) shock. We conducted a 2-phase, prospective, multicenter, randomized clinical trial. Patients with ischemic or nonischemic dilated cardiomyopathy and primary or secondary prevention indication for ICD were enrolled in an initial observational phase until first appropriate shock (phase A). After reconsenting, patients were randomly assigned 1:1 in phase B to immediate ablation (within 2 months from shock delivery) or continuation of standard therapy. The primary end point was a composite of death from any cause or hospitalization for worsening heart failure. Amiodarone intake was not allowed except for documented atrial tachyarrhythmias. On July 23, 2021, phase B of the trial was interrupted as a result of the first interim analysis on the basis of the Bayesian adaptive design. Of the 517 patients enrolled in phase A, 154 (30%) had ventricular tachycardia, 56 (11%) received an appropriate shock over a median follow-up of 2.4 years (interquartile range, 1.4-4.4), and 47 of 56 (84%) agreed to participate in phase B. After 24.2 (8.5-24.4) months, the primary end point occurred in 1 of 23 (4%) patients in the ablation group and 10 of 24 (42%) patients in the control group (hazard ratio, 0.11 [95% CI, 0.01-0.85]; P=0.034). The results met the prespecified termination criterion of >99% Bayesian posterior probability of superiority of treatment over standard therapy. No deaths were observed in the ablation group versus 8 deaths (33%) in the control group (P=0.004); there was 1 worsening heart failure hospitalization in the ablation group (4%) versus 4 in the control group (17%; P=0.159). ICD shocks were less frequent in the ablation group (9%) than in the control group (42%; P=0.039). Ventricular tachycardia ablation after first appropriate shock was associated with a reduced risk of the combined death or worsening heart failure hospitalization end point, lower mortality, and fewer ICD shocks. These findings provide support for considering ventricular tachycardia ablation after the first ICD shock. URL: https://www. gov; Unique identifier: NCT01547208.


Draft Genome Sequence of the Strict Anaerobe Clostridium homopropionicum LuHBu1 (DSM 5847).
Here, we report the draft genome sequence of Clostridium homopropionicum LuHBu1 (DSM 5847(T)), a strictly anaerobic bacterium, which performs propionate fermentation and is capable of growing with 2-, 3-, or 4-hydroxybutyrate as its sole substrate. The genome consists of a single chromosome of 3.65 Mb.