The Difference in Repeatability of Automated Superficial Retinal Vessel Density according to the Measurement Area Using OCT Angiography.
To evaluate the difference in the repeatability of automated superficial retinal vessel density and foveal avascular zone (FAZ) metrics according to the measurement area of optical coherence tomography angiography (OCTA). A total of 127 normal eyes from 127 healthy subjects were included. Macular angiography images were acquired from all subjects using the Zeiss Cirrus 5000 with AngioPlex™ OCTA software. Scans of 3 × 3 mm and 6 × 6 mm were each performed twice in a randomly arranged sequence. Vessel density (VD), perfusion density (PD), and FAZ metrics of the superficial capillary plexus were calculated automatically for all scans, and the repeatabilities for both scan patterns were assessed based on intraclass correlation (ICC), coefficient of variation (CV), and coefficient of repeatability (CR) parameters. The average measured values in the two scan patterns were also compared. VD was significantly greater in the 3 × 3 mm scan than in the 6 × 6 mm scan according to all parameters, whereas PD was significantly less in the 3 × 3 mm scan than in the 6 × 6 mm scan. The ICCs for VDs in the central fovea were 0.826 and 0.741 for the 3 × 3 and 6 × 6 mm scans, respectively, and the CVs were 8.00% and 12.75%. For PDs, the ICCs were 0.839 and 0.762 and the CVs were 9.32% and 14.90%. The FAZ metrics in the 3 × 3 mm scan showed good repeatability with an ICC >0.75 and a CV <10.0%. However, all ICCs for the 6 × 6 mm scans were <0.75, and the CVs were all >10%. The 6 × 6 mm macular angiography scans resulted in lower repeatabilities than the 3 × 3 mm scans according to all OCTA parameters, particularly in the central fovea and FAZ metrics. The 3 × 3 mm scan was more suitable than the 6 × 6 mm scan for analyzing macular microvascular density and FAZ metrics.


Re6C32: A Magnetic Pentagonal Icositetrahedron Molecule.
A chiral molecule with octahedral symmetry, Re6C32, has been identified using first-principles calculations. It is a hollow cage with the shape of Catalan pentagonal icositetrahedron. The calculated vibrational frequencies are in the range of 83.0-1341.2 cm-1, which indicate the stability of the Re6C32 molecule. Molecular dynamics simulations show that the topological structure of the Re6C32 molecule is well maintained up to 1500 K. The electronic structural analysis shows that there are significant p-d orbital hybridizations near the Fermi level. Moreover, the Re6C32 molecule is magnetic with spin magnetic moment of 12 μB. This magnetic carbon-based hollow Re6C32 cage may become a candidate for single-molecule devices.


Dynamics of lipids and polychlorinated biphenyls in a Baltic amphipod (Monoporeia affinis): a field study.
Processes such as accumulation and elimination, which control tissue concentration of polychlorinated biphenyls (PCBs), were examined over time in an in situ study of the amphipod Monoporeia affinis. These processes were studied with respect to individual PCB congeners, percentage lipid and composition, and body weight. A secondary objective was to examine the impact of seasonal variability in percentage lipids and lipid composition on PCB concentration in two coexisting Baltic amphipods, M. affinis and Pontoporeia femorata. Polycholorinated biphenyl concentrations tended to be higher in P. femorata than in M. affinis, possibly because of P. femorata's lower respiration rate and larger size. The net accumulation of PCBs was congener dependent and negatively correlated to lipid concentration. The relation between the net concentration change rate of 16 PCB congeners over time and log Kow was not significant during the spring and summer months, a time when lipid accumulation and strong growth occur. In contrast, the net concentration change rate of the corresponding PCB congeners over time during autumn and early winter, that is, from the period before gonad maturation to the period after mating and early embryogenesis, showed a significant relation to tog Kow (r2 = 0.62, p < 0.001, n = 16). During the latter period, amphipod lipid weight was reduced while the PCB body burden increased. Results strongly indicate that elimination rather than accumulation is the main process controlling amphipod body burden. This pattern results in a transfer of PCBs from the female to the developing embryos, which is reflected in high PCB body burden in newly released offspring.


Range-restricted, specialist Bornean butterflies are less likely to recover from ENSO-induced disturbance.
The forest fires induced by the El Niño Southern Oscillation (ENSO) in 1997-1998 resulted in the temporary extirpation of more than 100 lowland butterfly species at a forest site in Borneo. Species with more restricted ranges were less likely to recover over the following four years. Matched-pair analyses revealed that species with lower initial abundances, restricted geographic ranges, and more specialized larvae were less likely to return. Specialization differed predictably between the (more generalist) wide-range and (more specialized) restricted-range species in our data set, and both geographic range and level of specialization were important in multivariate models. These are the first observations directly linking extent of occurrence, ecological specialization, and observed recovery following local extirpation. If recovery time exceeds the frequency of disturbance, local extirpation can lead to local extinction. Given that ENSO-induced disturbances are increasing in frequency, in severity, and in geographic scale, these results suggest that specialist species with restricted geographic ranges could be at particularly high risk of global extinction.


Determination of nitrite in meat products and water using dapsone with combined spot test/diffuse reflectance on filter paper.
This paper describes a simple, portable and environmentally friendly procedure for the determination of nitrite by combined spot test/diffuse reflectance spectroscopy on filter-paper. The method is based on the diazotization reaction of dapsone (4,4'-diamino-diphenyl sulphone, DAP) and (naphthyl)ethylenediamine hydrochloride (NED) with nitrite in acidic medium, yielding a coloured compound on the surface of a filter paper. Experimental design methodologies were used to optimise the measurement conditions. The reflectometric measurements were carried out at 545 nm and the linear range obtained was 0.29-5.0 mg L(-1) of nitrite (R=0.997), with detection and quantification limits of 0.09 and 0.29 mg L(-1), respectively. The proposed method was applied successfully to the analysis of nitrite in meat products and well water samples.


Infrared optical absorption of magnetopolaron resonance states in graphene on the polar substrates.
We study the infrared optical absorption of magnetopolaron resonance states in graphene in the strong magnetic field based on the Huybrechts's model, in which polaron states are formed due to the strong coupling between electrons and surface optical (SO) phonons induced by the polar substrate. We propose the special magnetopolaron states [Formula: see text], namely, the superposition states between one SO phonon and the first-excited Landau level, which split into two branches of coupling modes and give rise to two optical absorption peaks with different intensities. Moreover, their intensities can be sensitively modulated by the magnetic field, the truncated wave-vector of SO phonon, polarity of substrate and internal distance between graphene and substrate. These results indicate that the structure of graphene laying on the polar substrate provide a good platform for exploring the polaron resonance states and magneto-optical transitions by infrared spectroscopy.


Relationship between epicardial adipose tissue attenuation and coronary artery disease in type 2 diabetes mellitus patients.
High epicardial adipose tissue (EAT) attenuation is a key characteristic of adipose tissue dysfunction and associated with coronary artery disease (CAD). As little is known about the modulation of EAT attenuation by metabolic disorders, we investigated the association between EAT attenuation and CAD risk factors, CAD presence and CAD severity in type 2 diabetes mellitus (T2DM) patients. We included 276 inpatients with T2DM and 305 control patients with normal glucose metabolism (NGM), who underwent cardiac computed tomography angiography (CCTA) and coronary artery calcium (CAC) scoring. EAT attenuation and volume were evaluated by contrast-enhanced CCTA image analysis. Furthermore, segment stenosis scores (SSSs) of the left main coronary artery (LMCA), left anterior descending artery (LAD), left circumflex artery (LCX), right coronary artery (RCA), diagonal/intermediate branch (D/I) and obtuse marginal branch (OM) were calculated to assess CAD severity. T2DM patients showed higher significant CAC scores, coronary plaque prevalence, total SSSs and LMCA-SSSs, LAD-SSSs, LCX-SSSs, RCA-SSSs and D/I-SSSs compared with NGM controls. In contrast to NGM controls, EAT volume was significantly increased in T2DM patients, whereas EAT attenuation was similar. In T2DM patients, EAT attenuation was associated with discrete CAD risk factors, the presence of coronary and triple-vessel plaques, as well as LAD-SSSs, LCX-SSSs, RCA-SSSs and total SSSs. In addition, EAT attenuation was only associated with the total SSS of calcified plaques, but not with noncalcified plaques. In T2DM patients, high EAT attenuation is associated with the presence and severity of CAD in general and with coronary stenosis caused by calcified plaques in particular.


EndoC-βH5 cells are storable and ready-to-use human pancreatic beta cells with physiological insulin secretion.
Readily accessible human pancreatic beta cells that are functionally close to primary adult beta cells are a crucial model to better understand human beta cell physiology and develop new treatments for diabetes. We here report the characterization of EndoC-βH5 cells, the latest in the EndoC-βH cell family. EndoC-βH5 cells were generated by integrative gene transfer of immortalizing transgenes hTERT and SV40 large T along with Herpes Simplex Virus-1 thymidine kinase into human fetal pancreas. Immortalizing transgenes were removed after amplification using CRE activation and remaining non-excized cells eliminated using ganciclovir. Resulting cells were distributed as ready to use EndoC-βH5 cells. We performed transcriptome, immunological and extensive functional assays. Ready to use EndoC-βH5 cells display highly efficient glucose dependent insulin secretion. A robust 10-fold insulin secretion index was observed and reproduced in four independent laboratories across Europe. EndoC-βH5 cells secrete insulin in a dynamic manner in response to glucose and secretion is further potentiated by GIP and GLP-1 analogs. RNA-seq confirmed abundant expression of beta cell transcription factors and functional markers, including incretin receptors. Cytokines induce a gene expression signature of inflammatory pathways and antigen processing and presentation. Finally, modified HLA-A2 expressing EndoC-βH5 cells elicit specific A2-alloreactive CD8 T cell activation. EndoC-βH5 cells represent a unique storable and ready to use human pancreatic beta cell model with highly robust and reproducible features. Such cells are thus relevant for the study of beta cell function, screening and validation of new drugs, and development of disease models.


Sequence variation in ROP8 gene among Toxoplasma gondii isolates from different hosts and geographical localities.
The protozoan parasite Toxoplasma gondii has a worldwide distribution; it can cause serious diseases in humans and almost all other warm-blooded animals. Different genotypes of T. gondii result in different lesions in the same host. T. gondii rhoptry protein 8 (TgROP8) is a major factor of T. gondii acute virulence. We examined sequence variation in the TgROP8 gene among T. gondii isolates from different hosts and geographical localities. The TgROP8 gene was amplified from individual isolates and sequenced. A phylogenetic tree was constructed using Bayesian inference, maximum parsimony, and maximum likelihood based on the sequences obtained plus TgME49 from the ToxoDB database. The TgROP8 gene was 1728 bp in length for all the examined T. gondii strains, and their A+T contents were 45.37-45.95%. Sequence analysis detected 140 (0.06-5.56%) variable nucleotide positions resulting in 96 (0-10.78%) amino acid substitutions. Sequence variations in the TgROP8 gene resulted in polymorphic restriction sites for endonucleases BstBI, BsaI, and XhoI, which allowed the differentiation of the three classical genotype strains (types I, II, and III) by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). However, phylogenetic analyses indicated that the TgROP8 gene is not a suitable genetic marker for population studies of T. gondii.


Contribution of mass spectrometry-based proteomics to immunology.
Antigen processing forwards various information about the cellular status and the proteome to the cell surface for scrutiny by the cellular immune system. Thus the repertoire of major histocompatibility complex (MHC)-bound peptides and the MHC ligandome, indirectly mirrors the proteome in order to make alterations instantly detectable and, if necessary, to oppose them. Mass spectrometry is the core technology for analysis of both proteome and MHC ligandome and has evoked several strategies to gain qualitative and quantitative insight into the MHC-presented peptide repertoire. After immunoaffinity purification of detergent-solubilized peptide-MHC complexes followed by acid elution of peptides, liquid chromatography-mass spectrometry is applied to determine individual peptide sequences and, thus, allow qualitative characterization of the MHC-bound repertoire. Differential quantification based on stable isotope labeling enables the relative comparison of two samples, such as diseased and healthy tissue. Targeted searches for certain natural ligands, such as the 'predict-calibrate-detect' strategy, include motif-based epitope prediction and calibration with reference peptides. Thus, various approaches are now available for exposing and understanding the intricacies of the MHC ligand repertoire. Analysis of differences in the MHC ligandome under distinct conditions contributes to our understanding of basic cellular processes, but also enables the formulation of immunodiagnostic or immunotherapeutic strategies.


Functional characterization of a glucosyltransferase gene, LcUFGT1, involved in the formation of cyanidin glucoside in the pericarp of Litchi chinensis.
Anthocyanins generate the red color in the pericarp of Litchi chinensis. UDP-glucose: flavonoid 3-O-glycosyltransferase (UFGT, EC. 188.8.131.52) stabilizes anthocyanidin by attaching sugar moieties to the anthocyanin aglycone. In this study, the function of an UFGT gene involved in the biosynthesis of anthocyanin was verified through heterologous expression and virus-induced gene silencing assays. A strong positive correlation between UFGT activity and anthocyanin accumulation capacity was observed in the pericarp of 15 cultivars. Four putative flavonoid 3-O-glycosyltransferase-like genes, designated as LcUFGT1 to LcUFGT4, were identified in the pericarp of litchi. Among the four UFGT gene members, only LcUFGT1 can use cyanidin as its substrate. The expression of LcUFGT1 was parallel with developmental anthocyanin accumulation, and the heterologously expressed protein of LcUFGT1 displayed catalytic activities in the formation of anthocyanin. The LcUFGT1 over-expression tobacco had darker petals and pigmented filaments and calyxes resulting from higher anthocyanin accumulations compared with non-transformed tobacco. In the pericarp with LcUFGT1 suppressed by virus-induced gene silencing, pigmentation was retarded, which was well correlated with the reduced-LcUFGT1 transcriptional activity. These results suggested that the glycosylation-related gene LcUFGT1 plays a critical role in red color formation in the pericarp of litchi.


Structuring functional groups of aquatic insects along the resistance/resilience axis when facing water flow changes.
Understanding how differences in intensity and frequency of hydrological disturbances affect the resistance and resilience of aquatic organisms is key to manage aquatic systems in a fast-changing world. Some aquatic insects have strategies that improve the permanence (resistance), while others use strategies that favor recolonization (resilience). Therefore, we carried out a manipulative experiment to understand the influence of functional characteristics of aquatic insects in their permanence and recolonization against hydrological disturbances in streams in the biodiversity hotspot of the Cerrado of Brazil. We placed 200 artificial substrates in five streams and submitted them to changing water flow regimes that differed both in frequency and intensity, and we observed the response of the aquatic community for 39 days. We used a hierarchical Bayesian approach to estimate the probabilities of permanence and recolonization of each life strategy group (nine groups). We observed that the most intense changes in the water flow tended to affect the permanence of almost all groups, but the intensity of this effect reduced over time. On the other hand, less frequent disturbances, regardless of intensity, tended to reduce the permanence of most groups of aquatic insects over time. The different effects of disturbance intensity may have been related to a greater recolonization capacity of some groups. The results we present are worrisome in a scenario of reduced riparian vegetation around streams and with the expectation of precipitation becoming more concentrated in shorter periods of time due to climate change in the Cerrado hotspot, reducing the occurrence of many groups of aquatic insects in their habitat, particularly those with traits associated with resistance against hydrological disturbance.


Milk whey protein decreases oxygen free radical production in a murine model of chronic iron-overload cardiomyopathy.
Chronic iron overload is a major cause of organ failure worldwide, but its pathogenesis remains to be elucidated. To examine in an experimental murine model of iron-overload cardiomyopathy the relation between milk whey protein and, first, the production of reactive oxygen free radical species and, second, antioxidant reserve status. B6D2F1 mice were randomly assigned to four treatment groups (n=8 per treatment group): placebo control; iron only; whey only; and iron with whey. Reactive oxygen free radical species in the heart were quantified by the cytotoxic aldehydes malondialdehyde (MDA), 4-hydroxy-nonenal (HNE) and hexanal, while antioxidant reserve status was quantified by glutathione (GSH) and glutathione peroxidase (GPx) activity in the heart tissue. Significantly decreased concentrations (pmol/100 mg wet weight tissue) of MDA (2468+/-261), HNE (912+/-38) and hexanal (5385+/-927) were observed in the heart tissue of the group receiving iron with whey, in comparison with the iron-only treatment group (MDA 9307+/-387, HNE 1416+/-157, hexanal 14,874+/-2955; P<0.001). Significantly increased GPx (141+/-38 IU/L) and GSH (521+/-136 IU/L) activity were observed in mice receiving iron with whey, in comparison with mice receiving iron only (GPx 100+/-10 IU/L, GSH 446+/-33 IU/L; P<0.001). Mice receiving iron treatments with whey supplementation had significantly lower concentrations of cytotoxic aldehydes and significantly higher cardiac levels of GPx and GSH activity than did iron-only treated mice. Additional basic research is warranted to examine the exact mechanisms by which milk whey protein protects the heart.


Temperature-dependent formation of a conjugate between tris(hydroxymethyl)aminomethane buffer and the malondialdehyde-DNA adduct pyrimidopurinone.
The stability of the major adduct formed between the endogenous product malondialdehyde (MDA) and deoxyguanosine, a pyrimidopurinone termed M1G-dR, was tested under a variety of conditions required for nucleic acid manipulation. M1G-dR was found to be stable at neutral pH and 37 degrees C but to be unstable when stored at -20 degrees C in the presence of Tris buffers. A new product with a characteristic absorption band at 350 nm was identified by 1H-NMR as an enamino-imine comprised of one molecule of Tris, one molecule of MDA, and deoxyguanosine. The formation of the conjugate was observed on reaction of Tris with M1G-dR or its ring-opened derivative N2-(3-oxo-1-propenyl)deoxyguanosine. The Tris-M1G-dR conjugate was unstable in aqueous solutions at room temperature, undergoing hydrolysis. However, the Tris conjugate of M1G base remained stable at room temperature in organic solvent. The isolation and properties of a conjugate between M1G-dR and Tris suggest that cross-links may form by reaction of MDA with DNA but they are likely to be unstable to hydrolysis.


[Occupational exposure to wood dust. Health effects and exposure limit values].
This article presents a review of the health effects of occupational exposure to wood dusts and of the data that could be used for setting occupational exposure limits for this nuisance. The causal role of wood dust in the onset of sinonasal cancers is solidly established by numerous epidemiological studies, and the magnitude of the risk is particularly high for adenocarcinoma induced by exposure to hardwood dust. However, no current data allows to rule out the carcinogenic role of softwood dusts and, in the view of protecting the health of the workers, it does not seem relevant to distinguish these two types of wood. Various impairments of the lung function have been frequently associated with exposure to both 'allergenic' and 'non-allergenic' wood dusts and may occur at very low concentrations. According to the SUMER 94 and CAREX studies, about 200 000 workers are currently exposed to wood dusts in France (about 1% of the working population between 1990 and 1994). When taking into account full professional careers, the percentage of workers having been occupationally exposed can be estimated to be about 15% for men and 5% for women. Measurements performed in France between 1987 and 2000 show that exposure levels are high, about 50% of the samplings being over 1mg/m(3) (actual TWA in France). Although the studies present limits, particularly for the quantitative assessment of individual exposure levels, it seems that nonmalignant effects are susceptible to arise at the level of 1mg/m(3); a limit value of 0.5mg/m(3) would credibly allow to protect exposed workers from most of the risks of nonmalignant pulmonary effects. However, it is impossible to assure that this value will avoid the induction of sinonasal cancer, even if this level is certainly lower than the levels to which the cases of sinonasal cancers published in the literature were exposed.


[Difficulties in diagnosing Marfan's syndrome in childhood and adolescence].
The diagnosis of Marfan's syndrome in childhood and adolescence is made by the criteria of the Gent nosology, which evaluates genetic data, family history and a spectrum of clinical criteria. Due to the age dependent manifestations of the clinical symptoms, combined with the extreme heterogeneity of Marfan's syndrome diagnosis in early childhood remains sometimes difficult. Prospectively, we analyzed the clinical symptoms of all patients where Marfan's syndrome was suspected. We evaluated those patients between January 1997 and April 2002 by an interdisciplinary approach. We compared the clinical datas of the patients by using the Gent nosology and the Berlin nosolgy. 34 patients underwent full follow-up. The median age was 10,32 years with a range of 0,01 to 37,31 years, 19 patients were male, 15 patients were female. In eight patients Marfan's syndrome could be rouled out, 9 of 26 patients (34,6 %) fullified the criteria of the Gent nosology, in 17 of 26 patients (65,4 %) Marfan's syndrome remained just suspected, but was not fullified by the criteria of the Gent nosology. Concerning the Berlin nosology 14 of 26 patients (53,8 %) fullified the criteria, 12 of 26 patients (46,2 %) failed. Due to the criteria of the Gent nosology 14 patients (53,8 %) fullified the criteria of skeletal involvement, 21 patients (80,8 %) fullified cardiovascular major manifestation, 6 patients (23,1 %) had an ophthalmic major manifestation, and 9 patients (34,6 %) had an affected first degree relative or were genetically determined. On the basis of the data of our patients the diagnosis of Marfan's syndrome in childhood and adolescence can be made more sensitive by the criteria of the Berlin nosology compared to the Gent nosology. This seems to be caused by the age dependent manifestations of the symptoms. Until diagnostic algorhythms of Marfan's syndrome in childhood remain suboptimal, continuous clinical follow-up for all cases even those only in the case of suspected Marfan's syndrome are necessary to exclude complicated course and to improve outcome.


MGDB: a comprehensive database of genes involved in melanoma.
The Melanoma Gene Database (MGDB) is a manually curated catalog of molecular genetic data relating to genes involved in melanoma. The main purpose of this database is to establish a network of melanoma related genes and to facilitate the mechanistic study of melanoma tumorigenesis. The entries describing the relationships between melanoma and genes in the current release were manually extracted from PubMed abstracts, which contains cumulative to date 527 human melanoma genes (422 protein-coding and 105 non-coding genes). Each melanoma gene was annotated in seven different aspects (General Information, Expression, Methylation, Mutation, Interaction, Pathway and Drug). In addition, manually curated literature references have also been provided to support the inclusion of the gene in MGDB and establish its association with melanoma. MGDB has a user-friendly web interface with multiple browse and search functions. We hoped MGDB will enrich our knowledge about melanoma genetics and serve as a useful complement to the existing public resources. Database URL: http://bioinfo.ahu.edu.cn:8080/Melanoma/index.jsp.


Physiological effect of circulating glucagon on the hepatic membrane potential.
The pancreatic hormone glucagon hyperpolarizes the liver cell membrane under various conditions. Here we investigated the physiological relevance of this effect by testing the influence of infusions of glucagon antiserum on the liver cell membrane potential in vivo. Intracellular microelectrode recordings of liver cells (up to 60/rat over 2 h) were done in anesthetized male rats. Livers were fixed in place, and recordings were done 10-30 min after intraperitoneal injections of glucagon or hepatic portal vein infusions of glucagon or specific polyclonal glucagon antibodies raised in rabbits. The isotonic lactose vehicle was used as a control for glucagon, and equal amounts of nonimmunized rabbit IgG were used as a control for glucagon antibodies. Intraperitoneal glucagon (400 microg/kg) hyperpolarized the liver cell membrane up to 12 mV, and intraportal glucagon (10 or 60 microg/kg) dose dependently hyperpolarized the liver cell membrane by 3-7 mV. Intraportal infusion of glucagon antiserum (in vitro binding capacity of 4 ng glucagon/rat) significantly depolarized the liver cell membrane by approximately 2.5 mV. The effects of both glucagon and glucagon antiserum reversed after 60-90 min. We conclude that glucagon is a physiologically important modulator of the liver cell membrane potential.


Bilingual word recognition in a sentence context.
This article provides an overview of bilingualism research on visual word recognition in isolation and in sentence context. Many studies investigating the processing of words out-of-context have shown that lexical representations from both languages are activated when reading in one language (language-non-selective lexical access). A newly developed research line asks whether language-non-selective access generalizes to word recognition in sentence contexts, providing a language cue and/or semantic constraint information for upcoming words. Recent studies suggest that the language of the preceding words is insufficient to restrict lexical access to words of the target language, even when reading in the native language. Eye tracking studies revealing the time course of word activation further showed that semantic constraint does not restrict language-non-selective access at early reading stages, but there is evidence that it has a relatively late effect. The theoretical implications for theories of bilingual word recognition are discussed in light of the Bilingual Interactive Activation+ model (Dijkstra and van Heuven, 2002).


A massively parallel pipeline to clone DNA variants and examine molecular phenotypes of human disease mutations.
Understanding the functional relevance of DNA variants is essential for all exome and genome sequencing projects. However, current mutagenesis cloning protocols require Sanger sequencing, and thus are prohibitively costly and labor-intensive. We describe a massively-parallel site-directed mutagenesis approach, "Clone-seq", leveraging next-generation sequencing to rapidly and cost-effectively generate a large number of mutant alleles. Using Clone-seq, we further develop a comparative interactome-scanning pipeline integrating high-throughput GFP, yeast two-hybrid (Y2H), and mass spectrometry assays to systematically evaluate the functional impact of mutations on protein stability and interactions. We use this pipeline to show that disease mutations on protein-protein interaction interfaces are significantly more likely than those away from interfaces to disrupt corresponding interactions. We also find that mutation pairs with similar molecular phenotypes in terms of both protein stability and interactions are significantly more likely to cause the same disease than those with different molecular phenotypes, validating the in vivo biological relevance of our high-throughput GFP and Y2H assays, and indicating that both assays can be used to determine candidate disease mutations in the future. The general scheme of our experimental pipeline can be readily expanded to other types of interactome-mapping methods to comprehensively evaluate the functional relevance of all DNA variants, including those in non-coding regions.


Therapeutic prospects of natural alpha interferon from normal human leucocytes in the treatment of genital condylomata in HIV positive women.
Various Authors underline that HIV positive women have a higher CIN incidence and a higher prevalence of HPV. HIV could act as cofactor in the proliferation of HPV manifestation, because of a hypothetical deficit of local immunity. 5 HIV positive patients have been examined: 3 of them presenting vulvocondylomas and/or VIN were treated with 1.000.000 IU alpha IFNn three times a week for three weeks. 2 of them presenting CIN and/or HPV were treated with a two-phase therapy: A) Induction therapy: 3.000.000 IU 3 times a week for three cycles. B) Maintenance therapy: 1.000.000 IU 3 times a week for three months. IFN was the primary treatment for viral lesions, even though an improvement of prognostic indices (CD4, CD4/CDB, 2M), has shown its usefulness in association or in alternative to AZT.


[Personality dynamics during prolonged lithium prevention].
The preventive lithium therapy of paroxysm-like endogenic psychoses is associated with a change in the personality responses, in particular to the patient's own mental disease which makes it possible to speak about certain personality changes that manifest themselves phenomenologically by the formation of the adequate criticism of the disease, the psychological dependence on the lithium therapy and by peculiar reduction of emotional responses. The pathogenetic treatment of the described phenomena is sophisticated and calls for the consideration of both therapy and features of the endogenous process.


The economic benefits of acetylcholinesterase inhibitors for patients with Alzheimer disease and associated dementias.
Most cost-effectiveness studies using simulation modeling have demonstrated that donepezil, rivastigmine, and galantamine are cost effective for the treatment of mild-to-moderate Alzheimer disease (AD). These conclusions are in large part based on the assumption that improvement in cognitive status, or prevention of cognitive and functional decline, reduces the amount of time patients spend institutionalized or receiving other full-time care. However, as discussed in this article, outcomes besides delay to institutionalization affect the costs of AD. In reviews of utilization data from Medicare and managed care organizations, it was noted that hospitalization and post acute care in skilled nursing facilities accounted for the largest amount of excess direct costs, even among patients with mild or moderate AD. These utilization reviews also suggest that many patients with AD and related dementias require inpatient care because they are not able to self-manage comorbid conditions. The improvements in cognitive status and daily functioning associated with acetylcholinesterase inhibitor (AChEI) therapy are expected to translate into improved management of comorbidities and reduced caregiver burden, thus reducing the total cost of care. To confirm these and other economic benefits of AChEIs, pharmacoeconomic outcomes should be evaluated routinely as part of randomized, controlled trials and through well-controlled observational studies of AD patients in community and institutional settings.


Soy β-conglycinin (7S globulin) reduces plasma and liver cholesterol in rats fed hypercholesterolemic diet.
The aim of this study was to examine the comparative hypocholesterolemic effect of soybean 7S fraction in rats fed a high-cholesterol diet. Soybean 7S globulin (β-conglycinin) was administered orally once a day to rats, and the effects were measured after 28 days. Wistar rats were divided into four groups: standard diet (STD) (casein alone), hypercholesterolemic (HC) diet (STD plus 1 g/100 g cholesterol and 0.5 g/100 g cholic acid), HC+7S(1) diet (HC diet plus 200 mg of 7S/kg of body weight/day), and HC+7S(2) diet (HC diet plus 300 mg of 7S/kg of body weight/day). Food intake, weight gain, animals' growth, and feeding efficiency ratio were similar among the STD and three HC groups, indicating that these parameters were not affected by treatments. Animals that had received different doses of soybean 7S globulin had lower total cholesterol (TC), triglycerides (TG), and low-density lipoprotein (LDL)/high-density lipoprotein (HDL) ratio in serum and lower levels of hepatic TC and TG than those fed only the HC diet. The atherogenic indexes of HC+7S(1) and HC+7S(2) groups were 40% and 55% lower than that of the HC group, respectively. The results showed that the oral daily administration of β-conglycinin in the diet to HC rats, at between 1.85% and 2.75% of total ingested protein, promotes the reduction of TC, LDL-cholesterol, and TG and an increase in HDL-cholesterol in the plasma, besides a small but significant reduction in cholesterol and TG levels in the liver of the animals as well as a reduced atherogenic index.


Growth and survival of Serratia marcescens under aerobic and anaerobic conditions in the presence of materials from blood bags.
Several patients receiving blood transfusions during the summer of 1991 developed bacteremia after the transfusion. In all cases, the infection was caused by Serratia marcescens. The same strain of Serratia marcescens was isolated from the patients and from the outer surface of unfilled blood bags. The transport containers for the blood bags were made anoxic by using a catalyst in order to prevent microbial growth. The survival and growth of S. marcescens K202, which was isolated from the blood bags, was studied at different oxygen concentrations in deionized water containing materials derived from the blood bags. The rate of survival and growth of S. marcescens was highest under anaerobic conditions, in which growth occurred with all materials and even in deionized water alone. In contrast, S. marcescens did not survive in control cultures under semi-anaerobic and aerobic conditions. Growth was observed, however, under both aerobic and semi-anaerobic conditions in the presence of each of the tested blood bag materials. These findings indicate that the conditions in the transport containers for the blood bags were favorable for the survival and growth of S. marcescens.


Novel dual inhibitors of 5-lipoxygenase and thromboxane A2 synthetase: synthesis and structure-activity relationships of 3-pyridylmethyl-substituted 2-amino-6-hydroxybenzothiazole derivatives.
As part of our search for novel antiinflammatory drug candidates, we have designed and synthesized a series of 3-pyridylmethyl-substituted 2-amino-6- hydroxybenzothiazoles. Introduction of a 3-pyridylmethyl group into the 2-amino group (type-A) or the benzene ring (type-B) of 2-amino-6-hydroxybenzothiazoles imparted dual inhibitory activity against the production by glycogen-induced peritoneal cells of rat (in vitro) of leukotriene B4 (LTB4) and thromboxane A2 (TXA2), while not significantly inhibiting that of prostaglandin E2 (PGE2). The observed inhibition of the former two arachidonic acid metabolites was indicated to be the result of a direct action on 5-lipoxygenase and TXA2 synthetase by a cell-free in vitro assay. On the other hand, the inhibitory activities against PGE2 production were for most compounds very weak, indicating that they did not inhibit cyclooxygenase. Structure-activity relationship studies concerning the position of the 3-pyridylmethyl group revealed that type-B compounds generally showed about 10-fold stronger inhibitory activity against TXA2 synthetase than type-A compounds. The position of the 3-pyridylmethyl group played an important role in TXA2 synthetase inhibition. When some of these compounds (8, 13a, 26a (E3040), 26b, 27b, and 28b) were orally administered in the rat TNB/ethanol-induced chronic colitis model (100 mg/kg), the production of both LTB4 and TXB2 in the rat colon was reduced (ex vivo). In addition, one type-B compound, 6-hydroxy-5,7-dimethyl-2-(methylamino)-4-(3-pyridylmethyl)benzothiazole (26a), demonstrated a therapeutic effect at treatments of 100 mg/kg po once daily for 11 days and showed almost comparable activity to sulfasalazine at a dose of 500 mg/kg, the reference drug for inflammatory bowel diseases, in this in vivo model.


Lipoma in the cerebellopontine angle--case report.
The authors report a case of a cerebellopontine (CP) angle lipoma with a very unusual histological appearance. The 38-year-old male patient suffered vertigo, left tinnitus, and left hearing disturbance. Computed tomography and magnetic resonance imaging showed a nonenhanced low-density area and a high-intensity region in the left CP angle, respectively. The tumor, which was only partially removed because of its tight adhesion to the VIIIth nerve and brainstem, consisted of mature lipocytes and contained a piece of cartilage, which is highly unusual.


Dimensional crossover and incipient quantum size effects in superconducting niobium nanofilms.
Superconducting and normal state properties of Niobium nanofilms have been systematically investigated as a function of film thickness, on different substrates. The width of the superconducting-to-normal transition for all films is remarkably narrow, confirming their high quality. The superconducting critical current density exhibits a pronounced maximum for thickness around 25 nm, marking the 3D-to-2D crossover. The magnetic penetration depth shows a sizeable enhancement for the thinnest films. Additional amplification effects of the superconducting properties have been obtained with sapphire substrates or squeezing the lateral size of the nanofilms. For thickness close to 20 nm we measured a doubled perpendicular critical magnetic field compared to its large thickness value, indicating shortening of the correlation length and the formation of small Cooper pairs. Our data analysis indicates an exciting interplay between quantum-size and proximity effects together with strong-coupling effects and the importance of disorder in the thinnest films, placing these nanofilms close to the BCS-BEC crossover regime.


Heat stress inducible cytoplasmic isoform of ClpB1 from Z. nummularia exhibits enhanced thermotolerance in transgenic tobacco.
Previously, we isolated CDS of Ziziphus nummularia isoform ZnJClpB1-C from heat stress-tolerant genotype Jaisalmer. To further functionally validate ZnJClpB1-C assumed function in tobacco and to generate novel germplasm for heat stress tolerance, this gene was transformed in the Nicotiana tabacum. ClpB proteins are the major key player required for basal and induced heat stress tolerance in plant cells under heat stress. In Ziziphus nummularia ClpB1-C transcript from genotype Jaisalmer was highly upregulated under heat stress conditions, as reported earlier. Nine transgenic lines (T1) from three transgenic tobacco events with single-copy integration (T0 stage) were taken for heat stress analysis at seedling stage. Mature tobacco transgenic plants did not show any deformity as compared to wild plants when grown under normal conditions. Overexpression of ZnJClpB1-C in tobacco significantly increased the tolerance to heat stress. Under heat stress conditions (42 °C), T1 transgenic tobacco seedlings showed higher photosynthetic rate, relative water content, membrane stability index and lower levels of MDA, compared to the wild type untransformed plants. The qRT-PCR analysis revealed different level of transgene expression (1.08 to 3.89 folds) in 9 T1 transgenic lines. In vitro roles of ZnJClpB1-C regulating thermotolerance is not reported so far. These results demonstrated the positive roles of ZnJClpB1-C in enhancing thermotolerance and its use as a genomic resource in the near future for developing heat stress-tolerant germplasm.


Luminescent transition metal complexes as sensors: structural effects on pH response.
A series of luminescent transition metal complexes using the pH-sensitive ligand 5-carboxy-1,10-phenanthroline has been synthesized and characterized. The complexes, based on Ru(II) and Re(I), show monotonic changes in both luminescent intensity and lifetime with pH values over the range 2 < pH < 9. The impact of various structural features on both the range of pH sensitivity and dynamic response was studied using both intensity and lifetime measurements. It was possible to predictably tune the pH sensitivity range over about 1.5 pKa units. While significant variation in the dynamic response range was observed, the correlation with structural features needs further study.


ROLE OF ANDROGEN RECEPTOR-TARGETED AGENTS IN LOCALIZED PROSTATE CANCER.
Anti-androgen therapy continues to be a basic pilar of treatment for both localized and metastatic prostate cancer. The advent of new generation of androgen receptor targeted agents (ARTA) transformed the care of patients with advanced disease. After such a success, the steps were taken to incorporate a new generation of ARTAs into the treatment landscape of localized prostate cancer. High-risk prostate cancer represents the most aggressive form of localized disease with significant metastatic potential and poor outcome. Here, the impact of novel therapies will likely be profound and transforming. This clinical space has already been a showcase for multidisciplinary treatment where the combination of local therapies with systemic treatment gradually improved patient outcomes and the chances of cure. The most recent step in redefining the treatment of localized disease is the adoption of novel ARTAs moving forward the multidisciplinary platform. In this narrative review, we discuss current clinical evidence supporting the use of novel ARTAs in patients with localized high-risk prostate cancer and cover recent developments in biomarker-driven strategies for treatment individualization in this clinical context.


Kinetics of Light-Induced Intramolecular Energy Transfer in Different Conformational States of NADH.
When bound to a protein, the coenzyme NAD+/NADH typically exists in an extended conformation, while in aqueous solutions it can be characterized by an equilibrium of folded and unfolded structures. It was recognized long ago that in the folded conformation light absorption at the adenine ring initiates an effective energy transfer (ET) toward the nicotinamide group, but the mechanism of this process is still unexplored. Here we apply ultrafast transient absorption measurements on NADH combined with compartmental model analysis for following the kinetics of the ET. We find that the actual ET is extremely rapid (∼70 fs). The high rate can be well described by a Förster-type mechanism, promoted by both the special photophysical properties of adenine and the subnanometer inter-ring distance. The rapid ET creates a vibrationally hot excited state on nicotinamide, the vibrational and electronic relaxation of which is characterized by 1.7 and 650 ps, respectively.


Single Incision Versus Conventional Laparoscopic Sleeve Gastrectomy for Morbid Obesity: A Meta-Analysis.
The purpose of this study was to review the existing evidence on obese patients treated with single-incision laparoscopic sleeve gastrectomy (SILSG) or conventional laparoscopic sleeve gastrectomy (LSG), to compare the perioperative parameters and outcomes of the two bariatric procedures. A systematic literature search was performed in PubMed, Scopus, and Cochrane library, in accordance with the PRISMA guidelines. Seventeen articles met the inclusion criteria and incorporated 3843 patients. This study reveals comparable mean operative time, length of hospital stay, and complications between the two approaches. The SILSG approach was associated with enhanced cosmetic results, but increased incisional hernia rate. These outcomes should be treated with caution given the small number of included comparative studies. Well-designed, randomized controlled studies, comparing LSG to SILSG, are necessary to assess further their clinical outcomes.


Four kinds of antibody positive paraneoplastic limbic encephalitis: A rare case report.
It is not uncommon to develop autoimmune encephalitis and paraneoplastic neurological syndromes (PNS). 4 kinds of antibody-positive autoimmune paraneoplastic limbic encephalitis (PLE) have not been reported. PNS are distant effects of cancer on the nervous system, rather than syndromes in which cancer directly invades and metastasizes to the nerves and/or muscle tissues. If the limbic lobe system of the brain is involved, this will result in PLE. The detection of patients with PNS is challenging since tumors that cause paraneoplastic neurologic disorders are often asymptomatic, obscure, and thus easily misdiagnosed or missed. Currently, single- or double-antibody-positive paraneoplastic marginal encephalitis has been reported. However, no cases of three or more-antibody-positive cases have been reported. Here, we report a case of PLE that is anti-collapsing response-mediator protein-5, anti-neuronal nuclear antibody-type 1, anti-aminobutyric acid B receptor, and anti-glutamate deglutase positive, and address relevant literature to improve our understanding of the disease. This article reports on the management of a case of PLE with four positive antibodies, a review of the literature, in order to raise awareness among clinicians.


The origins and early development of the ILAE/IBE/WHO global campaign against epilepsy: Out of the shadows.
The International League Against Epilepsy (ILAE)/International Bureau for Epilepsy (IBE)/World Health Organization (WHO) Global Campaign Against Epilepsy was launched in Geneva and Dublin in the summer of 1997. The second phase of the Campaign was launched by a major event in Geneva, led by WHO Director General Dr. Gro Harlem Brundtland in February 2001. Since then, the Campaign has been gathering momentum around the world culminating in the WHO General Assembly Resolution (WHA 68.20) on Epilepsy in May 2015 supported by 194 countries. Recently, the World Federation of Neurology and other neurological non-governmental organizations (NGOs) have joined forces with the Epilepsy Campaign, leading to the WHO General Assembly Resolution (WHA 73.10) in May 2022 promoting a 10-year Intersectoral Global Action Plan (IGAP) for Epilepsy and Other Neurological Disorders. I was privileged to serve as the first Chairperson of the Global Campaign Against Epilepsy and this year all my documents and correspondence relating to the Campaign have been delivered to the Wellcome Collection in London. These are the basis for this detailed account of the origins and early development of the Campaign. I describe the events leading to the birth of the concept, planning for the Campaign, the launch, development, and the achievements of phase one. This first phase focused on awareness raising, education, and involvement, especially within WHO, ILAE, and IBE, including a series of five Regional Public Health meetings and Declarations on Epilepsy. In 1999, the WHO raised the status of the Campaign to the highest level, the first ever for a Non-Communicable Disease, resulting in the high profile launch of phase two in 2001, paving the way to the continuing global momentum and achievements, including the 2015 and 2022 WHO Resolutions.


An investigation of CO2 laser scleral buckling using moiré interferometry.
To demonstrate suitability of moiré interferometry to assess and quantify laser-induced shrinkage of scleral collagen for buckling procedures. Scleral buckling of human cadaver eyes was investigated using a Coherent Ultrapulse CO2 laser. Projection moiré interferometry was employed to determine the out-of plane displacement produced by laser exposure, and in-situ optical microscopy of reference markers on the eye was used to measure in-plane shrinkage. Measurements based on moiré interferometry allow a three dimensional view of shape changes in the eye surface as laser treatment proceeds. Out-of-plane displacement reaches up to 1.5 mm with a single laser spot exposure. In-plane shrinkage reached a maximum of around 30%, which is similar to that reported by Sasoh et al (Ophthalmic Surg Lasers. 1998;29:410) for a Tm:YAG laser. The moiré technique is found to be suitable for quantifying the effects of CO2 laser scleral shrinkage and buckling. This can be further developed to provide a standardized method for experimental investigations of other laser sources for scleral shrinkage.


What Could Be Future Scenarios?-Lessons from the History of Public Health Surveillance for the Future: --A keynote address presented at the 8th World Alliance for Risk Factor Surveillance (WARFS) Global Conference on October 30, 2013, Beijing, China.
This article provides insights into the future based on a review of the past and present of public health surveillance-the ongoing systematic collection, analysis, interpretation, and dissemination of health data for the planning, implementation, and evaluation of public health action. Public health surveillance dates back to the first recorded epidemic in 3180 BC in Egypt. A number of lessons and items of interest are summarised from a review of historical perspectives in the past 5,000 years and the current practice of surveillance. Some future scenarios are presented: exploring new frontiers; enhancing computer technology; improving epidemic investigations; improving data collection, analysis, dissemination and use; building on lessons from the past; building capacity; and enhancing global surveillance. It is concluded that learning from the past, reflecting on the present, and planning for the future can further enhance public health surveillance.


Capillary leak syndrome after cardiopulmonary bypass in elective, uncomplicated coronary artery bypass grafting operations: does it exist?
Operations coupled with cardiopulmonary bypass may provoke a systemic inflammatory response, and it has been suggested that this responses causes capillary leakage of proteins, edema formation, and even organ failure. However, capillary leak syndrome is mainly a clinical diagnosis and has not been verified as yet by actual demonstration of protein leakage from the circulation. We have therefore measured the disappearance of labeled plasma protein before and after cardiopulmonary bypass. Sixteen patients scheduled for elective coronary artery bypass grafting were enrolled in a prospective controlled study. The cardiopulmonary bypass circuit was primed with crystalloids only. Tumor necrosis factor alpha, interleukin 6, interleukin 8, anaphylatoxin C3a, and terminal complement complex C5b9 levels were determined before, during, and 3 hours after cardiopulmonary bypass. The transvascular escape rate of plasma protein from the intravascular compartment was assessed by measuring the disappearance of intravenously injected Evans blue dye before and during the third hour after cardiopulmonary bypass. A significant inflammatory response could be demonstrated by means of the 5 measured mediators after bypass. The maximal increase, as compared with the baseline value, was found for interleukin 6 (36-fold). The transvascular escape rate of Evans blue dye was similar before and after bypass (7.6 +/- 0.6%/h vs 7.3 +/- 0.6%/h). The above data confirm the systemic inflammatory response induced by cardiopulmonary bypass. Contrary to expectations, the transvascular escape rate of Evans blue dye did not change when comparing values before and after bypass. The data do not support the concept of increased protein leakage in the exchange vessels after bypass. We were unable to demonstrate a capillary leak syndrome.


Evaluation of the effect of scleral buckling on the anterior chamber angle using ASOCT.
To evaluate the change in anterior chamber angle parameters after scleral buckling surgery using anterior segment optical coherence tomography (ASOCT). Fifty-five phakic eyes of 55 subjects undergoing scleral buckling with the placement of an encircling band for primary rhegmatogenous retinal detachment were included in this study. Anterior chamber angle assessment was performed using ASOCT. The trabecular iris angle (TIA), the angle-opening distance, and the trabecular iris space area were the parameters obtained preoperatively and 48 hours, 1 week, and 1 month after the surgery using ASOCT. The intraocular pressure (IOP) was measured using Goldmann applanation tonometry. The mean age of the patients was 43.2±11.3 years; there were 38 male and 17 female patients. There was a significant decrease in all the angle parameters (TIA, angle opening distance, and trabecular iris space area) at 48 hours, 1 week, and 1 month after surgery (P<0.0001). TIA 180 degree decreased from 41.93±14.27 degrees preoperatively to 31.09±10.52 degrees (P<0.0001) and TIA 0 degree from 42.7±14.04 degrees to 32.92±12.17 degrees (P<0.0001) at 48 hours postoperatively. The mean IOP at 48 hours (17.04±3.26 mm Hg) was significantly higher than the preoperative level (12.68±1.96 mm Hg; P<0.001). No significant difference was noted between the mean preoperative IOP and the IOP readings at 1 week (P=0.402) and 1 month (P=0.23) postoperatively. ASOCT imaging reveals that scleral buckling surgery for retinal detachment induces significant narrowing of the anterior chamber angle.


3'-(Phenyl alkynyl) analogs of abscisic acid: synthesis and biological activity of potent ABA antagonists.
We report here the synthesis and biological testing of 3'-(phenyl alkynyl) abscisic ABA analogs, a new class of potent ABA antagonists. These ABA analogs incorporate a rigid framework of eight carbon atoms attached at the 3'-carbon atom of ABA that prevents folding of the ABA analog-bound receptor required for ABA signalling. The two-step synthesis is based upon the optimized conversion of natural (S)-ABA to 3'-iodo ABA which can be coupled to phenyl acetylenes using Sonogashira conditions, or to styryl compounds through Suzuki chemistry. The parent 3'-(phenyl alkynyl) ABA analog 7 was obtained in 29% yield, 74% yield based on recovered starting material. In a lentil seed germination assay, compound 7 was found to have more potent activity than other known 3'-substituted ABA antagonists to date. In a structure activity study parasubstituted phenyl alkynyl analogs had comparable activity to the analog 7 while the 3'-styryl ABA 18 was only slightly less active. Analog 7 overcame ABA inhibition of germination and seedling growth in a wide range of mono and dicot plant species, including canola, lentil, soybean, rice, wheat, barley, cannabis and canary seed. 3'-(Phenyl alkynyl) ABA analogs have numerous potential practical agricultural applications including promoting ripening of crops, dormancy breaking of seeds and woody perennials, as well as promoting seed germination, and growth under stress conditions as demonstrated in this report.


Attitudes of adolescents towards sexual health in three cities from Croatia and Bosnia and Herzegovina.
This study aimed to determine the attitudes of adolescents towards sexual health and to assess their understanding of contraception use and knowledge about sexually transmitted infections in three ethnically different areas of Croatia and Bosnia and Herzegovina. A total of 146 adolescent high school students between 17 and 19 years of age from three different cities were enrolled in this cross-sectional study. The participants completed two anonymous questionnaires: one that measured attitudes towards safe sex practice and another that measured contraception and general sexual health knowledge. Shame was identified as the main factor why adolescents did not seek information about sexual health and contraception. On average, respondents correctly answered only 35% of questions regarding sexual health. Adolescents from East Mostar, with a predominantly Bosniak population, had more positive attitudes towards safe sex and showed less contraception knowledge in comparison to their colleagues from two cities in Croatia. Our study design might be burdened by socially desirable reporting and other biases inherent to the survey design. Adolescents from Croatia and Bosnia and Herzegovina, countries that experienced war and are still undergoing post-communist socio-economic transition, generally did not have adequate knowledge with respect to safe sex lifestyles. Respondents had major problems identifying sexually transmitted infections and their knowledge about contraception was insufficient. Providing care for adolescent populations should address specificities in local ethnical and socio-economical circumstances. There is a need for further studies to determine social, cultural and religious factors influencing the knowledge and attitudes towards sexual health in order to provide most effective interventions for specific adolescent populations.


Nanofiber alignment on a flexible substrate: hierarchical order from macro to nano.
A simplified approach to constructing a composite material comprised of aligned electrospun nanofibers onto a flexible substrate consisting of a microfilament yarn is presented. The metal-coated knit patterns of the microfilament yarn play the role of the parallel electrode, required for the alignment of electrospun nanofibers. Hybrid materials with knitted textile as a support material and aligned high-surface-area nanofibers could represent ideal materials for use in the filtration, optical, and biomedical industries.


A novel solid dosage form of rifampicin and isoniazid with improved functionality.
The aim of the present investigation was to develop a novel dosage form of rifampicin and isoniazid to minimize degradation of rifampicin in acidic medium and to modulate the release of rifampicin in the stomach and isoniazid in the intestine. Gastroretentive tablets of rifampicin (150 mg) were prepared by the wet granulation method using hydroxypropyl methylcellulose, calcium carbonate, and polyethylene glycol 4000. The granules and tablets of rifampicin were characterized. Hard gelatin capsules (size 4) containing a compacted mass of isoniazid (150 mg) and dicalcium phosphate (75 mg) were enteric coated. Two tablets of rifampicin and 1 capsule (size 4) of isoniazid were put into a hard gelatin capsule (size 00). The in vitro drug release and in vitro drug degradation studies were performed. Rifampicin was released over 4 hours by zero-order kinetics from the novel dosage form. More than 90% of isoniazid was released in alkaline medium in 30 minutes. The results of dissolution studies with the US Pharmacopeia XXIII method revealed that a substantial amount of rifampicin was degraded from the immediate release capsule containing rifampicin and isoniazid powder owing to drug accumulation in the dissolution vessel and also to the presence of isoniazid. The degradation of rifampicin to 3-formyl rifampicin SV (3FRSV) was arrested (3.6%-4.8% degradation of rifampicin at 4 hours) because of the minimization of physical contact between the 2 drugs and controlled release of rifampicin in acidic medium in the modified Rossett-Rice apparatus. This study concludes that the problem of rifampicin degradation can be alleviated to a certain extent by this novel dosage form.


VARIATION IN ENVIRONMENTAL AND GENOTYPIC SEX-DETERMINING MECHANISMS ACROSS A LATITUDINAL GRADIENT IN THE FISH, MENIDIA MENIDIA.
Models of environmental sex determination (ESD) usually assume that genetic influences on sex are polygenic, but the validity of this (or any other) form of genotype-environment interaction is virtually unknown. In the Atlantic silverside, Menidia menidia, sex is determined by an interaction between temperature and genotype and the response of sex ratio to temperature differs among populations from different latitudes. We examined the genetic basis of this pattern by measuring among family variation in the proportion of females, F/(F + M), within and among high (21°C) and low (15°C) temperatures for two populations: one from Nova Scotia (NS) where the level of ESD is low, and another from South Carolina (SC) where the level of ESD is high. In NS fish, temperature had a significant influence on sex ratio in only 1 of 23 families. The distribution of the fraction of females within temperatures for families from NS was highly heterogeneous and tended to fall into distinct classes (0.0, 0.25, 0.5, 1.0) like that expected from Mendelian segregation of a major sex factor(s). In contrast, temperature had a highly significant influence on sex ratio in all SC families examined (N = 24). Family sex ratios within temperatures were highly heterogeneous and, at least at 15°C, did not conform to simple Mendelian ratios. At 21°C, the proportion of females in most SC families was near zero and so the underlying sex tendencies of different families could not be discerned. Based on a previous study, mid-latitude fish appear to have an intermediate form of sex determination: simple Mendelian sex-ratio patterns exist and there is a moderate thermal influence on sex ratio in most but not all families. We suggest that sex determination in M. menidia is controlled by an interaction between major genetic factors, polygenic factors, and temperature and that the relative importance of each component differs with latitude. High latitude populations appear to have evolved a major sex-determining factor(s) that overrides the effect of temperature, and this factor(s) is lacking in low latitude populations.


Interdisciplinary approaches of transcranial magnetic stimulation applied to a respiratory neuronal circuitry model.
Respiratory related diseases associated with the neuronal control of breathing represent life-threatening issues and to date, no effective therapeutics are available to enhance the impaired function. The aim of this study was to determine whether a preclinical respiratory model could be used for further studies to develop a non-invasive therapeutic tool applied to rat diaphragmatic neuronal circuitry. Transcranial magnetic stimulation (TMS) was performed on adult male Sprague-Dawley rats using a human figure-of-eight coil. The largest diaphragmatic motor evoked potentials (MEPdia) were recorded when the center of the coil was positioned 6 mm caudal from Bregma, involving a stimulation of respiratory supraspinal pathways. Magnetic shielding of the coil with mu metal reduced magnetic field intensities and improved focality with increased motor threshold and lower amplitude recruitment curve. Moreover, transynaptic neuroanatomical tracing with pseudorabies virus (applied to the diaphragm) suggest that connections exist between the motor cortex, the periaqueductal grey cell regions, several brainstem neurons and spinal phrenic motoneurons (distributed in the C3-4 spinal cord). These results reveal the anatomical substrate through which supraspinal stimulation can convey descending action potential volleys to the spinal motoneurons (directly or indirectly). We conclude that MEPdia following a single pulse of TMS can be successfully recorded in the rat and may be used in the assessment of respiratory supraspinal plasticity. Supraspinal non-invasive stimulations aimed to neuromodulate respiratory circuitry will enable new avenues of research into neuroplasticity and the development of therapies for respiratory dysfunction associated with neural injury and disease (e.g. spinal cord injury, amyotrophic lateral sclerosis).


[Hemagglutination inhibition test for the diagnosis of coronavirus infection in cattle].
The main aim of the work was to obtain a viral corpuscle suspension with high hemagglutinating activity to be used as an antigen in HAI tests. The bovine coronavirus SC-1 strain was cultivated on ovine foetal renal cells and gave a suspension with a hemagglutinating titer of 1:64-1:128. A monospecific anti-coronavirus immune serum to be used in the HAI test, too, was prepared by hyperimmunization of a gnotobiotic calf. Its hemagglutinating and neutralizing titers were of 1:2560 and, respectively, 8.5 log. The HAI test was performed using the following reagents: the coronavirus suspension as a hemagglutinating antigen, a 1% suspension of mouse red blood corpuscles, the monospecific anti-coronavirus immune serum as a positive control, and a negative serum. 265 serum samples were investigated, collected from cows and calves in 18 farms where neonatal gastroenteritis was diagnosed. In 72.4% out of these samples there were positive results in the IHA test.


Intentional Replantation of an Avulsed Immature Permanent Incisor: A Case Report.
This case report discusses the successful endodontic treatment of an open apex maxillary right permanent central incisor that had been avulsed and incorrectly replanted in a 7-year-old patient. The tooth was carefully re-extracted followed by cleaning of the alveolar socket and immediate replantation. However, pulp necrosis was diagnosed, and regenerative endodontic treatment was performed. The root canal system was disinfected by passive ultrasonic irrigation with 2.5% sodium hypochlorite. At the first visit, the tooth was repositioned and immobilized with an appropriate semirigid splint. After 14 days, the splint was removed, and the diagnosis of pulp necrosis was confirmed by thermal testing. The root canal was emptied, disinfected, and filled with calcium hydroxide paste, which was left in place for 7 days. At the third visit, calcium hydroxide was removed with hand files and passive ultrasonic irrigation, and the canal was filled with a mixture of double antibiotic paste (metronidazole/ciprofloxacin) and zinc oxide. The antibiotic paste was left in place for 30 days. At the final visit, the paste was removed and the periapical area stimulated with a #80 K-file to encourage clot formation within the pulp cavity. A mineral trioxide aggregate paste cervical plug was placed, and the tooth was restored with glass ionomer cement. Clinical and imaging (radiographic and tomographic) follow-up at 3, 6, 12, and 36 months showed endodontic success with continued root formation.


Pt(0) and Pd(0) olefin complexes of the metalloid N-heterocyclic carbene analogues [E(I)(ddp)] (ddp=2-{(2,6-diisopropylphenyl)amino}-4-{(2,6-diisopropylphenyl)imino}-2-pentene; E=Al, Ga): ligand substitution, H--H and Si--H bond activation, and cluster formation.
Various products of the reaction of [E(ddp)] (ddp=2-{(2,6-diisopropylphenyl)amino}-4-{(2,6-diisopropylphenyl)imino}-2-pentene; E=Al, Ga) with Pt(0) and Pd(0) olefin complexes are reported. Thus, the reaction of [Pt(cod)(2)] (cod=1,5-cyclooctadiene) with two equivalents of [Ga(ddp)] yields [Pt(1,3-cod){Ga(ddp)}(2)] (1), whereas treatment of [Pd(2)(dvds)(3)] (dvds=1,1,3,3-tetramethyl1,3-divinyldisiloxane) with [E(ddp)] leads to the monomeric compounds [(dvds)Pd{E(ddp)}] (E=Ga (2 a), Al (2 b)) by substitution of the bridging dvds ligand. Both 1 and 2 a readily react with strong pi-acceptor ligands such as CO or tBuNC to give the dimeric compounds [M{mu(2)-Ga(ddp)}(L)] (L=CO, tBuNC; M=Pt (3 a, 5 a), Pd (3 b, 5 b)), respectively. Based on (1)H NMR spectroscopic data, [Pt{Ga(ddp)}(2)(CO)] is likely to be an intermediate in the formation of 3 a. Furthermore, reactions of 1 with H(2) and HSiEt(3) yield the monomeric compounds [Pt{Ga(ddp)}(2)(H)(2)] (7) and [Pt{Ga(ddp)}(2)(H)(SiEt(3))] (8). Finally, the reaction of [Pt(cod)(2)] with one equivalent of [Ga(ddp)] in the presence of H(2) in hexane gives the new dimeric cluster [Pt{mu(2)-Ga(ddp)}(H)(2)](2) (9).


Surface topography and ultrastructural architecture of the tegument of adult Carmyerius spatiosus Brandes, 1898.
Adult Carmyerius spatiosus or stomach fluke has an elongate, cylindrical-shaped, straight to slightly curved body, with conical anterior end and truncated posterior end. The worm measures about 8.7-11.2mm in body length and 2.3-3.0mm in body width across the mid-section. When observed by SEM, the tegumental surface in all part of the body appears highly corrugated with ridges and furrows, and having no spines. The ventral surface has more complex corrugation than those of the dorsal surface. Both anterior and posterior suckers have thick edges covered with transverse folds and appear spineless. The genital pore is located at the anterior part of the body. There are two types of sensory papillae on the surface: type 1 is bulbous in shape with nipple-like tips; type 2 has a similar shape with short cilia on the tip. The dorsal surface exhibits similar surface features, but papillae appear less numerous and are smaller. When observed by TEM, the tegument is divided into four layers. The first layer includes the ridges and furrows which are covered by a trilaminate membrane underlined by a dense lamina and coated externally with the glycocalyx. The second layer of the tegument is a narrow region of cytoplasm that contains high concentrations of ovoid electron lucent tegumental granules (TG1), and disc-shaped electron dense tegumental granules (TG2) as well as lysosomes. TG1 close to the surface invariably exocytose their content into bottoms of the ridges, while some TG2 are fused and have their membrane joined up with the surface membrane. The third layer is the widest middle area of the tegument which contains numerous and evenly distributed mitochondria. Both TG1 and TG2 granules are present but in much fewer number than in the first and second layers. The fourth layer is the innermost zone that rests on and couples with a thick basal lamina. The cytoplasm in this layer is loosely packed and contains numerous infoldings of the basal plasma membrane with closely associated mitochondria. It also contains fairly large numbers of TG1 and TG2 granules which are produced and transported to the tegument by one type of tegumental cells lying in rows underneath the muscular layers.


[Anatomy of pi in TCM].
Description about pi's (spleen) morphology, size, color and weight in Neijing and Nanjing conformed to that in anatomy. Most Chinese doctors inherited this opinion and it was also the mainstream viewpoint in TCM. While pi described by the famous doctor Wang Qingren actually was the pancreatic gland, which was not accepted by later generations. Zhang Shouyi, a doctor in modern times, thought pi in TCM should include the spleen and the pancreatic gland. This viewpoint was quoted and identified by many people, but it couldn't be right according to anatomy and research by the ancients. In TCM, people got anatomic knowledge in practice and pi should refer to the spleen in modern anatomy.


Recent advances in organic transistor printing processes.
Recent progress in organic field-effect transistor (OFET) printing processes is reviewed, and a perspective on the future of the field is discussed. The principles underlying the OFET printing techniques are introduced according to two categories: direct write printing and transfer printing. A comprehensive overview of the use of printing techniques in OFET production processes is also provided. Considerations for improving OFET device performance using printing processes are explored. Prior to OFET commercialization, the OFET printing techniques must satisfy several requirements, as discussed here.


Malignancies in the rheumatoid arthritis abatacept clinical development programme: an epidemiological assessment.
To provide context for the malignancy experience in the rheumatoid arthritis (RA) abatacept clinical development programme (CDP) by performing comparisons with similar RA patients and the general population. Malignancy outcomes included total malignancy (excluding non-melanoma skin cancer (NMSC)), breast, colorectal, lung cancers and lymphoma. Comparisons were made between the observed incidence in patients within the abatacept CDP and RA patients on disease-modifying antirheumatic drugs (DMARD) identified from five data sources: the population-based British Columbia RA Cohort, the Norfolk Arthritis Register, the National Data Bank for Rheumatic Diseases, the Sweden Early RA Register and the General Practice Research Database. Age and sex-adjusted incidence rates (IR) and standardised incidence ratios (SIR) were used to compare events in the abatacept trials with the RA DMARD cohorts and the general population. A total of 4134 RA patients treated with abatacept in seven trials and 41,529 DMARD-treated RA patients in the five observational cohorts was identified for study inclusion. In the abatacept-treated patients, the 51 malignancies (excluding NMSC), seven cases of breast, two cases of colorectal, 13 cases of lung cancer and five cases of lymphoma observed were not greater than the range of expected cases from the five RA cohorts. The SIR comparing RA patients with the general population were consistent with those reported in the literature. The IR of total malignancy (excluding NMSC), breast, colorectal, lung cancers and lymphoma in the abatacept CDP were consistent with those in a comparable RA population. These data suggest no new safety signals with respect to malignancies, which will continue to be monitored.


MITOCHONDRIAL DNA DIVERSITY IN THE SEA URCHINS STRONGYLOCENTROTUS PURPURATUS AND S. DROEBACHIENSIS.
Restriction-fragment analysis was used to measure mitochondrial DNA (mtDNA) variability in 79 individuals of two species of temperate sea urchins. For the purple urchin Strongylocentrotus purpuratus, individuals were collected 1,500 km apart in 1985 and again from the same localities in 1988 (about one urchin generation). Twenty mtDNA genotypes belonging to four clades were found among 38 individuals. All four clades were found in both localities and in both years. Genetic structure was further tested by calculating the degree of interdeme genetic variation (GST ) and comparing this value to the GST 's from randomly shuffled data. No geographic structure was found. For S. droebachiensis, only six mtDNA genotypes were found among 41 individuals collected from the Pacific and Atlantic oceans. More than 80% of the individuals belonged to two genotypes. The genotype that dominated collections in the Pacific also occurred in the Atlantic; however, a common Atlantic genotype was never found in the Pacific. These two genotypes were identical at 64 of 65 restriction sites, and were only 0.2% divergent from each other. GST analysis confirmed that there were significant genetic differences between Atlantic and Pacific populations. The small divergence between genotypes suggests recent, but not continuous, migration. These marine species show smaller genotypic differences than terrestrial species over similar spatial and temporal scales. Both recruitment of adults from planktonic larval pools and the spread of sibling larvae over large distances from parents probably act as buffers to genetic differences in species with planktonic life-history phases.


Intraosseous xanthoma associated with hyperlipoproteinemia. A case report.
Intraosseous xanthoma associated with hyperlipoproteinemia, a rare disorder, was observed in the entire distal femur of a 52-year-old man. Initial radiographs suggested a primary bone neoplasm; however, an open biopsy established a diagnosis of intraosseous xanthoma. Histologically, the lesion was characterized by replacement of normal bone and marrow with xanthoma cells and extracellular cholesterol clefts. The hyperlipidemia was classified as Type III hyperlipoproteinemia and was successfully controlled by dietary lipid restriction alone. The disability was effectively treated and ambulation was possible with the aid of a cane.


Emergence of compact structures in a Klein-Gordon model.
The Klein-Gordon model (KG) phi=P{'}(|phi|)phi/|phi| is Lorenz invariant and has a finite wave speed, yet its localized modes, whether Q balls or vortices, suffer from the same fundamental flaw as all other solitons-they extend indefinitely. Using the KG model as a case study, we demonstrate that appending the site potential, P{a}(phi|), with a subquadratic part P(|phi|)=b{2}|phi|{1+delta}+P{a}(|phi|), 0<or=delta<1, induces particlelike modes with strictly compact support. These modes are robust and shorten in the direction of motion. Their interactions, which occur only on contact, are studied in two and three dimensions and are shown to span the whole range from being nearly elastic to plastic.


An unusual radiological presentation of mucinous adenocarcinoma of the colon.
We report a case of an elderly patient presenting with the left iliac fossa mass. The provisional diagnosis included an inflammatory diverticular mass or sigmoid colon cancer. Interestingly, computed tomography (CT) of the abdomen and pelvis demonstrated the left incarcerated Spigelian hernia containing an inflamed loop of the colon with signs of an early strangulation. However, at operation, a mucinous tumor was found involving the descending and upper sigmoid colon. The tumor eroded through the anterior abdominal wall, which was excised "en bloc." In the presented case, CT findings suggestive of a benign etiology were misleading. This potentially could have had significant consequences if the patient was treated conservatively. This case highlights that clinical history and examination remain the core components of a safe surgical practice. Clinical judgment cannot be substituted even by the best quality imaging. Therefore, we feel that it is important to share our experience of the successful management of the presented case.


The relationship between hydration status, male sexual dysfunction and depression in hemodialysis patients.
Disturbances in sexual function are common among dialysis patients. Normal erections require a complex balance of physiological, psychological, emotional, hormonal, neurological and vascular factors. This study examined a possible association of overhydration (OH) with male sexual dysfunction and depression in hemodialysis (HD) patients. This cross-sectional study assessed hydration status by whole-body bioimpedance spectroscopy in patients on maintenance HD for more than 12 months. Patients were categorized according to OH to extracellular water (ECW) ratio: OH/ECW ratio >0.15 and OH/ECW ratio ≤0.15. Sexual function was assessed using the International Index of Erectile Function (IIEF) score. Psychological status was evaluated using the Beck Depression Inventory (BDI) score. Serum sex hormones were determined. Of 39 stable participants on HD, 53.8% were overhydrated (OH/ECW ratio >0.15) and 46.2% not overhydrated (OH/ECW ratio ≤0.15). Of participants with OH/ECW ratio >0.15, 85.7% had mild to severe ED, and 71.4% had abnormal BDI scores, ranging from mild mood disturbance to severe depression. Compared to patients with OH/ECW ratio ≤0.15, BDI scores, serum estradiol and plasma hsCRP were higher (18.48±8.34 vs 10.61±5.46, p<0.001; 140.10±44.51 vs 126.10±32.26, p=0.034; and, 17.70±12.14 vs 9.76±8.79, p=0.013; respectively) in those with OH/ECW ratio >0.15, while their IIEF score, serum total testosterone and dehydroepiandrosterone (DHEA) were lower (12.81±7.31 vs 41.44±23.79, p<0.001; 8.97±5.43 vs 14.10±8.30, p=0.013; and 85.31±55.14 vs 133.3±95.48, p=0.029; respectively). The OH/ECW ratio correlated inversely with the IIEF score (r=-0.69, p<0.001) and positively with BDI scores (r=0.64, p<0.001). IIEF scores were inversely correlated with BDI scores (r=-0.54, p<0.001). OH in HD patients was found to be associated with a higher prevalence of sexual dysfunction and depression, lower serum levels of total testosterone and DHEA, and higher levels of serum estradiol.


Impact of academic medical center access on outcomes in multiple myeloma.
Treatment at academic cancer centers (ACs) is associated with improved survival across hematologic malignancies, though the benefit in multiple myeloma (MM) has not been examined. This study aims to evaluate survival outcomes at Commission on Cancer accredited ACs compared to non-academic centers (NACs) for patients receiving MM-directed therapy. The National Cancer Database (NCDB) was used to identify demographics and overall survival (OS) of MM patients diagnosed from 2004 to 2017 and to compare outcomes by facility type. Survival analysis was repeated in a propensity score matched cohort, with NACs matched 1:1 to ACs by age, race, comorbidity score, insurance, year of diagnosis, distance traveled, and income. Of 163 375 MM patients, 44.5% were treated at ACs. Patients at ACs were more likely to receive MM-directed therapy compared to NACs (81% vs. 73%, p < .001). For patients receiving treatment, median OS at ACs was 71.3 months versus 41.2 months at NACs (p < .001). When adjusted for baseline demographics, patients treated at ACs had reduced mortality; hazard ratio (HR) 0.79 (95% CI 0.78-0.81, p < .001). The propensity score matched cohort maintained this survival benefit with a median OS of 59.9 months at ACs versus 37.0 months at NACs (p < .001), HR of 0.66 (95% CI 0.64-0.67, p < .001). ACs treated younger patients with fewer comorbidities and were more likely to treat racial minorities and patients with Medicaid or private insurance, and the uninsured. In this analysis, MM patients treated at ACs have significantly improved survival. While potentially related to access to specialized care, socioeconomic factors that drive facility selection may also contribute.


Lafoensia pacari extract inhibits IL-5 production in toxocariasis.
Toxocariasis is an infection induced by Toxocara canis, an intestinal parasite of dogs. In this study, an experimental murine model of toxocariasis was used to evaluate the anti-inflammatory activity of an ethanolic extract of Lafoensia pacari stem bark. Mice infected with T. canis were treated with L. pacari extract (200 mg/kg, p.o.). Subsequently, we observed a reduction in the number of eosinophils in the peritoneal cavity, bronchoalveolar fluid, blood and bone marrow. Production of interleukin (IL)-5, a major cytokine involved in eosinophilic differentiation, proliferation and activation, is also an important marker for infection. The reduced levels of IL-5 observed in serum, lung homogenates and bronchoalveolar fluid demonstrated the anti-inflammatory mechanisms of L. pacari. Larvae recovery from infected mice treated with L. pacari was comparable with that from untreated mice, suggesting that L. pacari is not toxic to the parasite. Nonetheless, our results demonstrate a potential therapeutic effect of L. pacari extract in IL-5-mediated inflammatory diseases and provide new prospects for the development of drugs to treat IL-5-dependent allergic diseases such as parasite infection and asthma.


Clinician perspectives of barriers in perinatal palliative care.
Perinatal palliative care (PPC) is a developing model of care aimed at providing supportive services to families anticipating fetal or neonatal demise. This study measured barriers physicians and advance practice nurses report in providing and referring patients to PPC. A cross-sectional survey design using the Perinatal Palliative Care Perceptions and Barriers Scale © was administered using a Web-based tool. Recruitment was completed via email and flyer invitations and list serves. Physicians (n = 66) and advance practice nurses (n = 146) participated. T-test and Mann-Whitney U were used to examine differences in clinician-reported barriers to PPC. Physicians and nurses differ significantly in the barriers they report. Nurses expressed more obstacles at the healthcare systems level reporting difficulty in their ability to garner interdisciplinary support and gain administrative backing. Physicians are more confident in their ability to counsel patients than nurses. Members of both disciplines express similar feelings of distress and helplessness when caring for families expecting a fetal or neonatal demise. They also report a lack of societal support and understanding about PPC. Cultivating an environment of collaboration and interdisciplinary communication can benefit both caregivers and patients. Nurses have an opportunity to lead and promote PPC endeavors through participating in advantageous partnerships and research. Both disciplines may benefit from interventions directed at increasing their comfort in caring for patients in a palliative setting through targeted education and supportive staff services.


Phosphonium-organophosphate ionic liquids as lubricant additives: effects of cation structure on physicochemical and tribological characteristics.
Our previous work suggested great potential for a phosphonium-organophosphate ionic liquid (IL) as an antiwear lubricant additive. In this study, a set of five ILs were carefully designed and synthesized, with identical organophosphate anions but dissimilar phosphonium cations, to allow systematic investigation of the effects of cation alkyl chain length and symmetry on physicochemical and tribological properties. Symmetric cations with shorter alkyl chains seem to increase the density and thermal stability due to closer packing. On the other hand, either higher cation symmetry or longer alkyl moieties induce a higher viscosity, though the viscosity index is dependent more on molecular mass than on symmetry. While a larger cation size generally increases an IL's solubility in nonpolar hydrocarbon oils, six-carbon seems to be the critical minimum alkyl chain length for high oil miscibility. Both the two ILs, that are mutually oil miscible, have demonstrated promising lubricating performance at 1.04% treat rate, though the symmetric-cation IL moderately outperformed the asymmetric-cation IL. Characterizations on the tribofilm formed by the best-performing symmetric-cation IL revealed the film thickness, nanostructure, and chemical composition. Results here provide fundamental insights for future molecular design in developing oil-soluble ILs as lubricant additives.


Functional characterization of a first avian cytochrome P450 of the CYP2D subfamily (CYP2D49).
The CYP2D family members are instrumental in the metabolism of 20-25% of commonly prescribed drugs. Although many CYP2D isoforms have been well characterized in other animal models, research concerning the chicken CYP2Ds is limited. In this study, a cDNA encoding a novel CYP2D enzyme (CYP2D49) was cloned from the chicken liver for the first time. The CYP2D49 cDNA contained an open reading frame of 502 amino acids that shared 52%-57% identities with other CYP2Ds. The gene structure and neighboring genes of CYP2D49 are conserved and similar to those of human CYP2D6. Additionally, similar to human CYP2D6, CYP2D49 is un-inducible in the liver and expressed predominantly in the liver, kidney and small intestine, with detectable levels in several other tissues. Metabolic assays of the CYP2D49 protein heterologously expressed in E. coli and Hela cells indicated that CYP2D49 metabolized the human CYP2D6 substrate, bufuralol, but not debrisoquine. Moreover, quinidine, a potent inhibitor of human CYP2D6, only inhibited the bufuralol 1'-hydroxylation activity of CYP2D49 to a negligible degree. All these results indicated that CYP2D49 had functional characteristics similar to those of human CYP2D6 but measurably differed in the debrisoquine 4'-hydroxylation and quinidine inhibitory profile. Further structure-function investigations that employed site-directed mutagenesis and circular dichroism spectroscopy identified the importance of Val-126, Glu-222, Asp-306, Phe-486 and Phe-488 in keeping the enzymatic activity of CYP2D49 toward bufuralol as well as the importance of Asp-306, Phe-486 and Phe-488 in maintaining the conformation of CYP2D49 protein. The current study is only the first step in characterizing the metabolic mechanism of CYP2D49; further studies are still required.


WNT10B mutations associated with isolated dental anomalies.
Isolated hypodontia is the most common human malformation. It is caused by heterozygous variants in various genes, with heterozygous WNT10A variants being the most common cause. WNT10A and WNT10B are paralogs that likely evolved from a common ancestral gene after its duplication. Recently, an association of WNT10B variants with oligodontia (severe tooth agenesis) has been reported. We performed mutational analysis in our cohort of 256 unrelated Thai families with various kinds of isolated dental anomalies. In 7 families afflicted with dental anomalies we detected 4 heterozygous missense variants in WNT10B. We performed whole exome sequencing in the patients who had WNT10B mutations and found no mutations in other known hypodontia-associated genes, including WNT10A, MSX1, PAX9, EDA, AXIN2, EDAR, EDARADD, LPR6, TFAP2B, LPR6, NEMO, KRT17, and GREM2. Our findings indicate that the variants c.475G>C [p.(Ala159Pro)], found in 4 families, and c.1052G>A [p.(Arg351His)], found in 1 family, are most probably causative. They also show that WNT10B variants are associated not only with oligodontia and isolated tooth agenesis, but also with microdontia, short tooth roots, dental pulp stones, and taurodontism.


[Characteristics of peripheral blood leukocyte differential counts in patients with COVID-19].
To investigate the early changes of peripheral blood leukocyte differential counts in patients with COVID-19. Ten patients with COVID-19 and 30 patients with other viral pneumonia (non-COVID-19) admitted to The Sixth People's Hospital of Shanghai and Jinshan Branch Hospital from January 22 to February 17, 2020 were enrolled in this study. The differential counts of white blood cells (WBC) were analyzed. Patients in COVID-19 group showed relatively lower absolute WBC count 4.95(3.90,6.03)×10(9)/L, lymphocyte absolute count 1.20(0.98,1.50)×10(9)/L and eosinophil absolute count 0.01(0.01,0.01)×10(9)/L. Leukopenia developed in two patients(2/10), lymphocytopenia also in two patients(2/10). Seven over ten patients presented with eosinophil cytopenia. In non-COVID-19 group, absolute WBC count was 8.20(6.78,9.03)×10(9)/L (P<0.001), lymphocyte absolute count 1.75(1.20,2.53)×10(9)/L(P=0.036), eosinophil absolute count 0.02(0.01,0.03)×10(9)/L(P=0.005). Lymphocytopenia occurred in 16.7% patients, eosinophil cytopenia in 16.7% patients too. In conclusion, leukopenia, lymphocytopenia and eosinophil cytopenia are more common in COVID-19 patients than those in non-COVID-19 patients.


Nevirapine resistance in women and infants after first versus repeated use of single-dose nevirapine for prevention of HIV-1 vertical transmission.
Single-dose (SD) nevirapine (NVP) significantly reduces mother-to-child transmission of human immunodeficiency virus (HIV). We analyzed NVP resistance after receipt of SD NVP in 57 previously SD NVP-naive women, in 34 SD NVP-experienced women, and in 17 HIV-infected infants. The proportion of women infected with variants with resistance mutations, the types of mutations detected, and the frequency and level of K103N were similar in the two groups of women at 6 weeks and 6 months post partum. NVP resistance was detected in a similar proportion of infants born to SD NVP-naive versus SD NVP-experienced women. Repeated use of SD NVP to prevent HIV transmission does not appear to influence NVP resistance.


The Integrative Regulatory Network of circRNA, microRNA, and mRNA in Atrial Fibrillation.
Atrial fibrillation (AF) is the most common irregular heart rhythm which influence approximately 1-2% of the general population. As a potential factor for ischemic stroke, AF could also cause heart failure. The mechanisms behind AF pathogenesis is complex and remains elusive. As a new category of non-coding RNAs (ncRNAs), circular RNAs (circRNAs) have been known as the key of developmental processes, regulation of cell function, pathogenesis of heart diseases and pathological responses which could provide novel sight into the pathogenesis of AF. circRNAs function as modulators of microRNAs in cardiac disease. To investigate the regulatory mechanism of circRNA in AF, especially the complex interactions among circRNA, microRNA and mRNA, we collected the heart tissues from three AF patients and three healthy controls and profiled their circRNA expressions with circRNA Microarray. The differentially expressed circRNAs were identified and the biological functions of their interaction microRNAs and mRNAs were analyzed. Our results provided novel insights of the circRNA roles in AF and proposed highly possible interaction mechanisms among circRNAs, microRNAs, and mRNAs.


Characterization of a marsupial sperm protamine gene and its transcripts from the North American opossum (Didelphis marsupialis).
A synthetic oligonucleotide primer, designed from marsupial protamine protein-sequence data [Balhorn, R., Corzett, M., Matrimas, J. A., Cummins, J. & Faden, B. (1989) Analysis of protamines isolated from two marsupials, the ring-tailed wallaby and gray short-tailed opossum, J. Cell. Biol. 107] was used to amplify, via the polymerase chain reaction, protamine sequences from a North American opossum (Didelphis marsupialis) cDNA. Using the amplified sequences as probes, several protamine cDNA clones were isolated. The protein sequence, predicted from the cDNA sequences, consisted of 57 amino acids, contained a large number of arginine residues and exhibited the sequence ARYR at its amino terminus, which is conserved in avian and most eutherian mammal protamines. Like the true protamines of trout and chicken, the opossum protamine lacked cysteine residues, distinguishing it from placental mammalian protamine 1 (P1 or stable) protamines. Examination of the protamine gene, isolated by polymerase-chain-reaction amplification of genomic DNA, revealed the presence of an intron dividing the protamine-coding region, a common characteristic of all mammalian P1 genes. In addition, extensive sequence identity in the 5' and 3' flanking regions between mouse and opossum sequences classify the marsupial protamine as being closely related to placental mammal P1. Protamine transcripts, in both birds and mammals, are present in two size classes, differing by the length of their poly(A) tails (either short or long). Examination of opossum protamine transcripts by Northern hybridization revealed four distinct mRNA species in the total RNA fraction, two of which were enriched in the poly(A)-rich fraction. Northern-blot analysis, using an intron-specific probe, revealed the presence of intron sequences in two of the four protamine transcripts. If expressed, the corresponding protein from intron-containing transcripts would differ from spliced transcripts by length (49 versus 57 amino acids) and would contain a cysteine residue.


[Tobacco consumption and behavior of the smokers in Jiangxi province].
To explore the behaviors of smokers as well as the tobacco consumption in Jiangxi province. Impact regarding the increases of tobacco retail price on the smoking behaviors among cigarette consumers was also studied. Data was provided for further development on measurements including legislation, taxing and increasing the retail price on tobacco. 805 residents above 15 year-olds were chosen through random clustered sampling, and face to face interviewed, in Shangrao and Yichun of Jiangxi province. A chart was drawn to show the relationship between the change of smoking behavior and the increase of sales price of tobacco. The average rate of smoking was 27.08% (46.62% for males and 3.05% for females). 16.06% (35/218) of the smokers had ever tried to quit smoking but failed. The years of smoking ranged from 1 year to 50 years, with 96.33% of the smokers purchased cigarettes on their own. Price of the cigarette ranged from 15 to 30 RMB per package, and smokers usually spent on average 192 RMB which accounted 13% of their monthly income, per month. According to the chart, if the cigarette price had a 50% increase, it would lead to 45% of the smokers reduce the number of cigarette consumption with another 5% of the smokers attempt to quit smoking. If cigarette price had an increase by 2 times, 50% of the smokers would attempt to quit smoking. So the chart could be used to estimate the percentage of smokers' behavior change according to the increase of tobacco's sales price. The economy burden of tobacco consumption among cigarette smokers was huge, suggesting that the Chinese smokers did need to change their smoking behavior. Chinese government should develop tobacco-related legislation and laws and set proper cigarette tax, hopefully this could reduce the number of smokers in a long run.