Preliminary evaluation of a dual chamber pacemaker with bradycardia diagnostic functions.
Unexplained syncope is the main indication for the implantation of a diagnostic pacemaker. Studies on those implanted have shown that in patients with unexplained syncope, the diagnosis of paroxysmal bradycardia was feasible and reliable. The present study was designed to evaluate a new bradycardia diagnosis algorithm, loaded in a dual chamber pacemaker, in 24 patients considered as candidates for diagnostic pacemakers. During a mean follow-up of 153 days, at least one bradycardia episode was recorded in 13 patients. The mean number of detected bradycardias was 6 and the median was 3. The mean delay between the algorithm activation and the first bradycardia episode was 67 days. The mechanism of bradycardia was atrioventricular block in 6 patients, sinus node dysfunction in 6 patients, and consecutive blocked atrial premature beats in 1 patient, as indicated by the event markers. In 11 patients bradycardia was recorded during the daytime only or day and night. In two patients the episodes were recorded only at night. Overall, the algorithm was well-tolerated; however, some mild symptoms were observed due to the method of bradycardia determination, allowing bradycardia. Three patients were symptomatic as a direct result of the algorithm operation, and four patients had symptoms related to the single chamber operation of the pacemaker while functioning in the diagnosis mode (VDI). These symptoms were relieved with DDD pacing.


State-of-the-art-myocardial perfusion stress testing: Static CT perfusion.
Large multicenter studies and meta-analysis have documented the diagnostic accuracy and the prognostic implications of stress echocardiography, cardiac magnetic resonance and, mainly, nuclear stress tests. However, none of them provides a comprehensive anatomical and functional evaluation within the same study as stress CT perfusion. Myocardial CT perfusion is the only non-invasive modality that allows to quantifying coronary stenosis and determining its functional relevance, constituting a potential "one-stop-shop" method for the diagnosis and global management of patients with known or suspected coronary artery disease. In comparison with the dynamic modality, that requires increased radiation, precise acquisition protocols and dedicated post-processing softwares, static CT perfusion was associated with less radiation exposure, non-inferior diagnostic accuracy, easier interpretation of images and is nowadays more widely available.


Transcytosis of IgG anti-D by human term trophoblast cells in culture.
Placental trophoblast cells were cultured on filters that allow access to medium bathing the apical and basal surfaces of cells. Purified human IgG was added to either the apical or the basal chambers and sampled at intervals of up to 100min for determination of IgG concentration by ELISA. Using this experimental system, transport of IgG1 and IgG3 monoclonal anti-D, affinity-purified polyclonal anti-D and human polyclonal IgG were shown to occur primarily in the apical to basal (i.e. maternal to fetal) direction. The overall transport of monoclonal and polyclonal anti-D was less than 10% in 45 min, several times lower than that of IgG. There was no difference in the rate or percentage of transport between IgG1 (BRAD-5) and IgG3 (BRAD-3) monoclonal anti-D. The possibility that the Fc receptor mediating transcytosis of IgG anti-D through human trophoblast cells in culture is the placental hFcRn is proposed.


Self-directed assembly of photoactive hybrid silicates derived from an azobenzene-bridged silsesquioxane.
Hybrid silicate materials derived from organo-bridged silsesquioxane precursors, RO3-Si-R'-Si-OR3, where R and R' are organic ligands, represent a remarkably diverse class of nanocomposites capable of forming both Si-O-Si and Si-C-Si bonds with molecular scale homogeneity. Recently, in an effort to better control their structure and function, surfactant-directed self-assembly or self-directed assembly has been used to synthesize hierarchical organo-bridged polysilsesquioxanes that exhibit order over multiple length scales. Here we report the synthesis and self-directed assembly of an optically active azobenzene-bridged silsesquioxane, 4,4'-bis(3-triethoxysilylpropylureido)azobenzene 1. Hydrogen-bonding interactions between the three active centers of the bis-ureide groups (-NH-CO-NH) combined with pi-pi interactions between the azobenzene groups serve to self-assemble 1 into a highly ordered lamellar mesostructure in which the d-spacing is optically controlled through photoisomerization of the azobenzene moiety before or after assembly.


Estimation of genetic diversity in a natural population of cambui tree (Myrciaria tenella O. Berg) using ISSR markers.
Cambui (Myrciaria tenella O. Berg) is a native species from Brazil, which belongs to the family Myrtaceae. Molecular characterization is one of the most used tools for the study of the biotechnological potential of species because the diversity level between individuals can be inferred. Analysis of genetic diversity is fundamental to the direction of the strategies necessary to form and maintain a germplasm. This study aimed to evaluate the genetic diversity in a natural population of cambui using inter-simple sequence repeat (ISSR) molecular markers. The natural population, which provided the plant material, is found at the Private Reserve of Natural Heritage of Caju, which belongs to the experimental field of Embrapa Tabuleiros Costeiros, in the municipality of Itaporanga d'Ajuda, SE, Brazil. Young leaves of each individual were collected for DNA extraction and analysis of PCR-ISSR. Thirty primers were tested and the top 10 were selected. The use of these primers resulted in 71 fragments with 98.3% polymorphism. Similarity of individuals ranged between 0.30 and 0.92. The most similar individuals were C13 and C17 and the most distant were C1 and C41. Through UPGMA, six distinct groups were identified. This information may be used for conservation of these genetic resources, germplasm exchange, creation of germplasm bank and in future studies with this species.


Measurement of nicotine intake in pregnant women--associations to changes in blood cell count.
Self-reported information about smoking is imprecise and subject to bias, with accuracy varying according to circumstances. Biochemical assessment gives much clearer indications of the effects of tobacco intake on physiological parameters. As part of a randomized controlled trial, a new point-of-care test for smoking was used as a tool to reduce smoking in pregnancy. Measurements of nicotine metabolites in urine were related to a physiological effect of smoking, notably changes to blood parameters, assessed as a routine part of antenatal care. One hundred and eighty-seven pregnant women attending outpatient antenatal care were initially questioned by a midwife about smoking habits and later questioned by the investigators, during which the test was performed and the results relayed back to the patient. Self-reported smoking habit and cigarette consumption, either reported to midwives or the investigators, were shown to be poor indicators of the effects of smoking on blood parameters. The biochemical assessment of nicotine intake was significantly related to white blood count, haemoglobin concentration, haematocrit, mean cell volume, and mean cell haemoglobin. Red cell count, mean cell haemoglobin concentration, and platelet count were unrelated to nicotine metabolite measurements. We concluded that the new test was a reliable measure of nicotine intake in pregnancy and the results correlated with smoking-related changes to haematological parameters.


Reversal of P-glycoprotein-mediated multidrug resistance by diallyl sulfide in K562 leukemic cells and in mouse liver.
Multidrug resistance (MDR) mediated by the overexpression of drug efflux protein P-glycoprotein (P-gp) is one of the major obstacles to successful cancer chemotherapy. P-gp acts as an energy-dependent drug efflux pump, reducing the intracellular concentration of structurally unrelated drugs. Modulators of P-gp function can restore the sensitivity of multidrug-resistant cells to such drugs. In the present study, we evaluated the P-gp modulatory potential of diallyl sulfide (DAS), a volatile organosulfur compound present in garlic, known to possess many medicinal properties, including antimutagenic and anticarcinogenic activities. For in vitro studies, K562 leukemic cells were made resistant (K562/R) to the cytotoxicity of vinblastine (VBL) by progressive adaptation of the sensitive K562 parental cells to VBL. Cross-resistance of K562/R was found between vincristine (VCR), doxorubicin and other antineoplastic agents. A non-toxic concentration of DAS (8.75 x 10(-3) M) enhanced the cytotoxic effects of VBL and another vinca alkaloid, VCR, time dependently in VBL-resistant human leukemia (K562/R10) cells but had no effect on the parent (K562/S) cells. The results show that DAS decreased the induced levels of P-gp in resistant cells back to the normal levels as analyzed both qualitatively and quantitatively by western blotting and immunocytochemistry. Furthermore, in vivo combination studies showed that DAS effectively inhibited vinca alkaloid-induced P-gp overexpression in mouse hepatocytes. Quantitation of immunostained tissue sections with image analysis showed that the reduction in P-gp levels was up to 73% for VBL- and 65% for VCR-induced drug resistance. The above features thus indicate that DAS can serve as a novel, non-toxic modulator of MDR and can be used as a dietary adjuvant.


Visualizing polynucleotide polymerase machines at work.
The structures of T7 RNA polymerase (T7 RNAP) captured in the initiation and elongation phases of transcription, that of phi29 DNA polymerase bound to a primer protein and those of the multisubunit RNAPs bound to initiating factors provide insights into how these proteins can initiate RNA synthesis and synthesize 6-10 nucleotides while remaining bound to the site of initiation. Structural insight into the translocation of the product transcript and the separation of the downstream duplex DNA is provided by the structures of the four states of nucleotide incorporation. Single molecule and biochemical studies show a distribution of primer terminus positions that is altered by the binding of NTP and PP(i) ligands. This article reviews the insights that imaging the structure of polynucleotide polymerases at different steps of the polymerization reaction has provided on the mechanisms of the polymerization reaction. Movies are shown that allow the direct visualization of the conformational changes that the polymerases undergo during the different steps of polymerization.


A survey on high-concentration-capability headspace sampling techniques in the analysis of flavors and fragrances.
This survey critically discusses high-concentration-capacity (HCC) headspace (HS) techniques applied to sample the volatile fraction of matrices of interest in the flavors and fragrance fields. In particular, the advantages, limits, and fields of application of HS solid-phase microextraction (SPME), high-capacity HS sorptive extraction (HSSE) and HS solid-phase dynamic extraction (SPDE) are evaluated. These techniques are discussed in view of the peculiar characteristic of HCC-HS techniques, from the standpoint that these techniques are a bridge between static (S-HS) and dynamic HS (D-HS) because they are as simple, fast, easy to automate, and reliable as S-HS, yet afford analyte concentration factors comparable to those of D-HS. Moreover, the different degree of their development is a consequence of the different times in which they were introduced into the market, because the potential of HS-SPME is now well known, having been introduced approximately 12 years ago, but that of HSSE has still to be fully explored, and HS-SPDE still has to be investigated because it is very recent.


A towards-multidimensional screening approach to predict candidate genes of rheumatoid arthritis based on SNP, structural and functional annotations.
According to the Genetic Analysis Workshops (GAW), hundreds of thousands of SNPs have been tested for association with rheumatoid arthritis. Traditional genome-wide association studies (GWAS) have been developed to identify susceptibility genes using a "most significant SNPs/genes" model. However, many minor- or modest-risk genes are likely to be missed after adjustment of multiple testing. This screening process uses a strict selection of statistical thresholds that aim to identify susceptibility genes based only on statistical model, without considering multi-dimensional biological similarities in sequence arrangement, crystal structure, or functional categories/biological pathways between candidate and known disease genes. Multidimensional screening approaches combined with traditional statistical genetics methods can consider multiple biological backgrounds of genetic mutation, structural, and functional annotations. Here we introduce a newly developed multidimensional screening approach for rheumatoid arthritis candidate genes that considers all SNPs with nominal evidence of Bayesian association (BFLn > 0), and structural and functional similarities of corresponding genes or proteins. Our multidimensional screening approach extracted all risk genes (BFLn > 0) by odd ratios of hypothesis H1 to H0, and determined whether a particular group of genes shared underlying biological similarities with known disease genes. Using this method, we found 6614 risk SNPs in our Bayesian screen result set. Finally, we identified 146 likely causal genes for rheumatoid arthritis, including CD4, FGFR1, and KDR, which have been reported as high risk factors by recent studies. We must denote that 790 (96.1%) of genes identified by GWAS could not easily be classified into related functional categories or biological processes associated with the disease, while our candidate genes shared underlying biological similarities (e.g. were in the same pathway or GO term) and contributed to disease etiology, but where common variations in each of these genes make modest contributions to disease risk. We also found 6141 risk SNPs that were too minor to be detected by conventional approaches, and associations between 58 candidate genes and rheumatoid arthritis were verified by literature retrieved from the NCBI PubMed module. Our proposed approach to the analysis of GAW16 data for rheumatoid arthritis was based on an underlying biological similarities-based method applied to candidate and known disease genes. Application of our method could identify likely causal candidate disease genes of rheumatoid arthritis, and could yield biological insights that not detected when focusing only on genes that give the strongest evidence by multiple testing. We hope that our proposed method complements the "most significant SNPs/genes" model, and provides additional insights into the pathogenesis of rheumatoid arthritis and other diseases, when searching datasets for hundreds of genetic variances.


A novel instrument to determine pulp proximity.
The aim was to determine whether extracted teeth could be used to test the Prepometer instrument, which indicates pulp proximity with green/amber/red light emitting diodes. Third molars were reduced to a plane in dentine and Prepometer readings made. Dentine was removed in 0.5 mm increments and readings made until only lights 9 or 10 (red) showed. The teeth were sectioned and the dentine thickness measured. Analysis permitted construction of a highly significant predictor-model (p < 0.01), the red/amber light boundary coinciding with a dentine thickness of 2.4 mm. The Prepometer was consistent in predicting pulp proximity but was more sensitive than specified.


Contribution of alpha2beta1, alpha3beta1, alpha6beta4 integrins and 67 kDa laminin receptor to the interaction of epidermoid carcinoma A-431 cells with laminin-2/4.
Laminin-2/4 is the major laminin isoform of normal muscle and nerve tissues and plays an important role in tumor invasion and metastasis. Despite the fact that laminin-2/4 has been found in the skin basement membrane, insufficient evidence is available on the effect of laminin-2/4 on the behavior of both normal and transformed skin cells. A comparison of the contribution of alpha2beta1, alpha3beta1, alpha6beta4 integrins and 67 kDa laminin receptor on the surface of the human epidermoid carcinoma cell, A-431, to interaction with laminin-2/4 was carried out. The cell interaction with extracellular matrix component is a multistage process. We employed new methods for studying different stages of the interaction of A-431 cells with laminin-2/4. We demonstrated that integrins alpha2beta1, alpha3beta1, alpha6beta4 and 67 kDa laminin receptor are involved in the interaction of A-431 cells with laminin-2/4. We found that contribution of the same receptors to different stages of the interaction with laminin can be different. alpha2beta1 integrins are involved in EGF-induced A-431 cells' migration on laminin-2/4. We demonstrated the cooperation between alpha2beta1 and alpha3beta1 integrins during adhesion and spreading of A-431 cells on laminin-2/4-coated substrate. These results provide information about laminin-2/4 receptors and their contribution to different stages of the interaction with cells.


Murine monoclonal antibodies (MCS-1 and MCS-2) reactive with human myeloid leukemia cells.
In an attempt to identify the antigens expressed on human myeloid leukemia cells, two murine monoclonal antibodies (mAb) designated as MCS-1 (isotype; IgG3) and MCS-2 (IgG1) were raised. MCS-1 reacted with peripheral blood granulocytes, but not with monocytes, whereas MCS-2 reacted with both granulocytes and monocytes. The incidence of MCS-1 and MCS-2 reactivity with the cells from a total of 121 patients with various type of leukemias was as follows: 25/46 (54%) and 39/47 (83%) in acute myeloblastic leukemia and acute monoblastic leukemia, 8/16 and 16/16 in chronic myeloid leukemia in blastic crisis (CML-BC) of myeloid type, 0/7 and 2/7 in CML-BC of lymphoid type, 0/26 and 2/32 in acute lymphoblastic leukemia (ALL), 7/7 and 7/7 in chronic phase of CML, respectively. Two cases of MCS-2 positive ALL had a Philadelphia chromosome marker (Ph1). In contrast, neither MCS-1 nor MCS-2 reacted with lymphocytes or any of leukemic and non-leukemic lymphoid cell lines tested. These results indicate that MCS-1 and MCS-2 mAb recognized two different differentiation antigens expressed on granulocytes and monocytes. These mAb would be useful reagents to determine differentiation antigens on the cells in granulocyte and monocyte lineage.


Long-term effect of postnatal alcohol exposure on the number of cells in the neocortex of the rat: a stereological study.
Behavioral and morphological studies suggest that exposure to alcohol during development may cause damage in the neocortex. In this study, rat pups were exposed to alcohol during the brain growth spurt and examined at adulthood to ascertain the long-term effect of alcohol exposure on the neocortex. Four-day-old rat pups were surgically implanted with an intragastric cannula while under ether anesthesia and artificially reared from postnatal day (PN) 4 through PN11. Two of the consecutive 12 daily feeds contained either alcohol (4.5 g/kg; alcohol-exposed) or an isocaloric maltose/dextrin solution (gastrostomy control) from PN4 through PN9. On PN115, animals were perfused intracardially and the brains removed. Unbiased stereological methods were used to determine the neocortical volume, the total number of neurons and glial cells in the entire neocortex and in layer V, and the mean cell volume of neurons or mean nuclear volume of glial cells in layer V. No effect of alcohol was seen in the neuronal population on either cell number or mean cell volume, nor was there any difference in the total number or mean nuclear volume of glial cells in layer V. These findings suggest that neither the entire neocortex nor layer V alone are vulnerable to permanent alcohol-induced cell death.


Osteoporosis syndrome in thalassaemia major: an overview.
Osteoporosis in thalassaemia major (TM) represents a prominent cause of morbidity. The mechanism of pathogenesis of bone disease (BD) in TM is multifactorial and complicated. Peak bone mass is achieved shortly after completion of puberty and normally remains stable until the third decade of life when age-related bone mass begins. Growth hormone (GH) and sex steroids play a crucial role in bone remodeling and in the maintenance of skeletal architecture during adult life. GH and insulin growth factors (IGFs) have anabolic effect in bone formation. Sex steroids act probably by increasing the expression of RANKL by osteoblastic cells and alterations in the RANK/RANKL/OPG system in favor of osteoclasts. Impaired GH secretion and lack of sex steroids in thalassemic patients due to pituitary damage, contribute to failure of achieving optimal peak bone mass. Other endocrine complications such as hypoparathyroidism and vitamin D deficiency have also a detrimental role on bones in TM. It is still questionable whether the international criteria for defining osteopenia and osteoporosis are relevant to patients with TM; also a question arises for the diagnostic methods such as DEXA scan and management of osteoporosis with known treatment protocols, in the thalassaemic patient.


PACT, a double-stranded RNA binding protein acts as a positive regulator for type I interferon gene induced by Newcastle disease virus.
Virus infection triggers innate responses to host cells including production of type I interferon (IFN). Since IFN production is also induced by treatment with poly(I:C), viral double-stranded (ds) RNA has been postulated to play a direct role in the process. In the present study, we investigated the effect of dsRNA binding proteins on virus-induced activation of the IFN-beta gene. We found that PACT, originally identified as protein activator for dsRNA-dependent protein kinase (PKR) and implicated in the regulation of translation, augmented IFN-beta gene activation induced by Newcastle disease virus. Concomitantly with the augmented activity of IFN-beta enhancer, increased activity of NF-kappaB and IRF-3 and IRF-7 was observed. For the observed effect, the dsRNA-binding activity of PACT was essential. We identified residues of PACT that interact with a presumptive target molecule to exert its function. Furthermore, PACT colocalized with viral replication complex in the infected cells. Thus the observed effect of PACT is novel and PACT is involved in the regulation of viral replication and results in a marked increase of cellular IFN-beta gene expression.


Homovanilic acid in Huntington's disease and Sydenham's chorea.
Homovanilic acid (HVA) was determined in the lumbar CSF of 12 patients with Huntington's disease and 12 with Sydenham's chorea before and after probenecid administration. The means of HVA concentration (basal and after probenecid) were lower in those with Huntington's disease than in controls, and were even lower in a sub-group characterised by increased tone and slowness of voluntary movement. There was no correlation between CSF HVA values and the severity of abnormal movements, nor with length of the illness and age of the patients with Huntington's disease. The mean basal HVA concentration did not differ from controls in those with Sydenham's chorea but the accumulation with probenecid was significantly lower. These results suggest a decrease in cerebral dopamine release in both forms of chorea.


Exploring Perceived and Objective Measures of the Neighborhood Environment and Associations with Physical Activity among Adults: A Review and a Meta-Analytic Structural Equation Model.
There is an urgent need to understand factors that promote physical activities (PA) because it is one of the modifiable risk factors for global mortality. None of the previous reviews considered both perceived and objective characteristics of the same environment. The first aim was to review the literature on studies investigating the relationship between PA behavior of adults and perceived and objective physical environment measures. The second aim was to verify the potential mediational role of the perceived measure in the relationship between the objective measure of the environment and PA using meta-analytic SEM. Only 15 studies reported a relationship between PA and both environmental measures. One of the most studied characteristics of the physical environment is the accessibility to recreational/PA facilities. Both objective and subjective measures of accessibility to PA facilities are associated with PA. Meta-SEM results suggest a significant effect of the objective accessibility to facilities on PA behavior (β = 0.15) and on the perceived measure (β = 0.10), but the indirect effect was not significant. No significant effect was found for the perceived measure on PA, suggesting that individuals' level of awareness about their environments may have played a role. This prompts a need to create awareness campaigns.


Candidatus Rickettsia colombianensi in ticks from reptiles in Córdoba, Colombia.
Wildlife animals are reservoirs of a large number of microorganisms pathogenic to humans, and ticks could be responsible for the transmission of these pathogens. Rickettsia spp. are the most prevalent pathogens found in ticks. This study was conducted to detect Rickettsia spp. in ticks collected from free-living and illegally trafficked reptiles from the Department of Córdoba, Colombia. During the period from October 2011 to July 2014, ticks belonging to the family Ixodidae were collected, preserved in 96% ethanol, identified using taxonomic keys, and pooled (between 1 and 14 ticks) according to sex, stage, host, and collected place for subsequent DNA extraction. Rickettsia detection was performed using real-time polymerase chain reaction (RT-PCR), followed by conventional PCR to amplify a larger fragment of the gltA and 16S rRNA genes. The amplicons were sequenced using the Sanger method, and the nucleotide sequences were subjected to BLAST analysis to identify homologous sequences in GenBank, after which phylogenetic analysis was performed using the MEGA X software. In total, 21 specimens of nine species of reptiles were sampled, from which 805 Amblyomma dissimile ticks were collected, but only 180 ticks were selected to create 34 groups. The DNA of Rickettsia spp. was detected in 30/34 (88%) groups. The sequences of the gene gltA and 16S rRNA revealed a 100% identity with Candidatus Rickettsia colombianensi (GenBank: KF905456 and GenBank: KF691750). A. dissimile was the only tick found in all the sampled reptiles. The presence of Candidatus Rickettsia colombianensi in reptile ticks could represent a public health problem due to the risk of transmission to humans and the introduction of microorganisms to other geographical areas.


Natural product discovery through microbial genome mining.
The advent of the genomic era has opened up enormous possibilities for the discovery of new natural products. Also known as specialized metabolites, these compounds produced by bacteria, fungi, and plants have long been sought for their bioactive properties. Innovations in both DNA sequencing technologies and bioinformatics now allow the wealth of sequence data to be mined at both the genome and metagenome levels for new specialized metabolites. However, a key problem that remains is rapidly and efficiently linking these identified genes to their corresponding compounds. Within this review, we provide specific examples of studies that have used the power of genomic or metagenomic data to overcome these problems and identify new small molecules and their biosynthetic pathways.


Determination of the benzodiazepine plasma concentrations in suicidal patients using a radioreceptor assay.
Impairments in memory and psychomotor function appear to be induced by benzodiazepines not only after long-term use, but also after administration of a single dose. Because it is known on which neurotransmitter system the benzodiazepines exert their action, the use of a quantitative radioreceptor assay (RRA) can be a useful tool in studying the interrelationship between the neurochemical and memory processes. The RRA measures the sum of the main compound(s) and all active metabolites present, where it relates the biological activity to the pharmacodynamic effect instead of relating it to the plasma levels of the individual compounds. To correlate the loss of memory with the benzodiazepine concentration, plasma concentrations were determined in suicidal patients. From suicidal patients (n = 84), the benzodiazepines in plasma were measured with a direct radioreceptor assay using tritiated flunitrazepam as the labelled ligand. The receptor material was a lyophilized preparation from calf cortex. Furthermore, the samples were subjected to high-performance liquid chromatographic (HPLC) analysis, and the HPLC data were converted to diazepam equivalents using cross-reactivities of the individual compounds. Patients who had ethanol residues in their plasma were excluded from this correlation experiment. The data (n = 40) obtained with the two analytical techniques were compared and correlated to assess the validity of the radioreceptor assay in establishing the relationship between the loss of memory and the total amount of benzodiazepines present. The cumulative amount of diazepam determined with the RRA and the sum of compounds determined with the HPLC method, after correction using the cross-reactivities, were plotted and correlated using regression analysis. Regression analysis showed an x variable of 0.75 and a correlation coefficient of 0.67. The intercept was not significantly different from zero (P = 0.49, t-test), whereas the slope was significantly different from zero (P < 0.01). Benzodiazepines can be directly determined in plasma using this radioreceptor assay. The data obtained from HPLC analysis were easily converted to diazepam equivalents using the cross-reactivities. A discrepancy between the data obtained from the two analytical techniques, however, indicates that certain metabolites are present, which were not quantitated in the HPLC analysis, but were measured in the radioreceptor assay. Therefore, the radioreceptor assay proved to be a valuable tool for the assessment of clinical effects, such as the demonstration of the loss of memory in suicidal patients after a benzodiazepine overdose.


Rapunzel Alopecia: A Peculiar Form of Non-Marginal Traction Alopecia Secondary to Excessively Long Hair.
Traction alopecia (TA) is an underreported scarring alopecia in children and adolescents. TA occurs from chronic tensile forces on the hair and can be caused by different hairstyles that pull the hair tightly, such as regularly wearing braids, ponytails, cornrows, dreadlocks, and weaves, or by treating hair with rollers, hair extensions, or chemical relaxers.


Synthesis of Monodisperse Silica Particles Grafted with Concentrated Ionic Liquid-Type Polymer Brushes by Surface-Initiated Atom Transfer Radical Polymerization for Use as a Solid State Polymer Electrolyte.
A polymerizable ionic liquid, N,N-diethyl-N-(2-methacryloylethyl)-N-methylammonium bis(trifluoromethylsulfonyl)imide (DEMM-TFSI), was polymerized via copper-mediated atom transfer radical polymerization (ATRP). The polymerization proceeded in a living manner producing well-defined poly(DEMM-TFSI) of target molecular weight up to about 400 K (including a polycation and an counter anion). The accurate molecular weight as determined by a GPC analysis combined with a light scattering measurement, and the molecular weight values obtained exhibited good agreement with the theoretical values calculated from the initial molar ratio of DEMM-TFSI and the monomer conversion. Surface-initiated ATRP on the surface of monodisperse silica particles (SiPs) with various diameters was successfully performed, producing SiPs grafted with well-defined poly(DEMM-TFSI) with a graft density as high as 0.15 chains/nm². Since the composite film made from the silica-particle-decorated polymer brush and ionic liquid shows a relatively high ionic conductivity, we have evaluated the relationship between the grafted brush chain length and the ionic conductivity.


In situ XRD studies of nanocrystallization of Fe-based metallic glass: a comparative study by reciprocal and direct space methods.
Structural changes occurring in an Fe(72.5)Cu1Nb2Mo2Si(15.5)B7 alloy during a combination of constant rate heating (20 K min(-1)) and isothermal holding at 500 and 520 °C were investigated using in situ high-energy X-ray diffraction. We found that the ferromagnetic-to-paramagnetic transition of the amorphous phase is revealed as a change in the slope of the thermal expansion curve when heating a sample at a constant rate up to 520 °C. Real space analysis by means of the atomic pair distribution function (PDF) demonstrated that the rate and extent of the thermal expansion strongly depend on the interatomic separation. The PDF proved to be a reliable method for the description of crystallization kinetics. Further it allows determination of sizes of ultrafine nanocrystals with grain sizes well below 8 nm and thus makes observation of early stages of nanocrystallization possible. Following grain growth kinetics during isothermal annealing at 500 and 520 °C we found that the activation energy of the process is 357 ± 12 kJ mol(-1).


White collar stress: what studies of physicians can teach us.
The impact of occupational stress on physicians and their medical practice has been subject to much debate but relatively little research. This paper reviews some recent studies concerning the psychosocial work environment of physicians. Psychiatric and cardiovascular epidemiologic data indicate that physicians are at similar or enhanced risk to socioeconomically comparative groups. Clinical studies of physicians reveal associations between psychosocial work characteristics and traditional cardiovascular risk factors. Further work is needed to enhance our understanding of the implication of present work environments on the health and well-being of physicians as well as other health care workers.


Negative Pressure Wound Therapy Becomes the Treatment of Choice of Deep Sternal Wound Infection.
Sternal wound infection, especially deep sternal wound infection, is a serious complication after open heart surgery. It leads to a marked increase in hospital stay, financial expenses, and mortality. Treatment is primarily surgical and may be divided into conventional treatment methods and negative pressure wound therapy. Between 2010 and 2021, 77 patients presenting back after cardiac surgery with deep sternal wound infection were treated surgically. Conventional treatment methods were utilized in 45 patients and included wound revision with primary closure, continuous wound irrigation, and open treatment with secondary closure. Negative-pressure wound therapy (NPWT) was applied in 32 patients. The two treatment arms were compared by two primary outcomes - rate of recurrent infection and hospital mortality. Predictors of mortality and infectious recurrence were identified using multivariate logistic regression. Recurrent infection occurred in 18.2% of cases and mortality was 13% in the whole group. NPWT was more successful in preventing recurrent infection OR: 5.4 (95% CI: 1.1-27.5; P = 0.044) than conventional treatment and more than moderate left ventricular systolic dysfunction (EF<40%) predisposed to infectious recurrence - OR: 4.7 (95% CI: 1.05-22.1; P = 0.049). Recurrent infection itself was the strongest predictor of mortality in the multivariate model OR: 0.14 (95% CI: 0.03 - 0.58; P = 0.007). NPWT as an initial method of wound preconditioning followed by definitive wound closure effectively reduces the rate of infectious recurrence and patient mortality. It may become the modality of first choice when dealing with complicated incisional infections following heart surgery.


[Adult bronchiectasis revealing familial ciliary anomaly].
We report a case of bronchiectasis in a 26-year-old man associated with the following congenital abnormalities: deafness, purulent bronchorrhea, nasal polyps, dysmorphic physical pattern and chronic sinusitis. Situs inversus was absent. A sampling was performed on the posterior nasal mucous membrane and displayed structural ciliary abnormality: a deficiency of the intern dynein-arm. The patient's bother was affected and had similar features: congenital bronchiectasis, deafness, mental deficiency and sinusitis. Young's syndrome was relevant in this case. Hereditary ciliary dyskinesia should be considered in adults with bronchiectasis together with rhinologic and alimentary canal disorders. Nasal biopsies are safe and allow cilia examination.


Hsa-miR-375 promotes the progression of inflammatory bowel disease by upregulating TLR4.
To elucidate the biological function of hsa-miR-375 in the progression of inflammatory bowel disease (IBD) and the potential mechanism. Intestinal mucosa tissues of 26 IBD patients and 30 healthy volunteers who underwent colonoscopy were harvested for determining hsa-miR-375 level by quantitative Real-time polymerase chain reaction (qRT-PCR). Binding of hsa-miR-375 to toll-like receptor 4 (TLR4) was verified by the dual-luciferase reporter gene assay. Changes in the viability and apoptosis in Caco-2 cells influenced by hsa-miR-375 were examined by cell counting kit-8 (CCK-8) assay and flow cytometry, respectively. The regulatory effect of hsa-miR-375 on the intestinal epithelial barrier was examined by detecting transepithelial electrical resistance (TEER) and lucifer yellow flux. Relative levels of TLR4, nuclear factor-kappa B (NF-κB), zonula occludens-1 (ZO-1), occludin and inflammatory factors in Caco-2 cells were detected by qRT-PCR, Western blot and enzyme-linked immunosorbent assay (ELISA). Hsa-miR-375 was downregulated in intestinal mucosa tissues of patients with Crohn's disease (CD) and ulcerative colitis (UC). Knockdown of hsa-miR-375 decreased viability and TEER, but elevated apoptotic rate and lucifer yellow flux. Overexpression of hsa-miR-375 achieved the opposite trends. TLR4 was the direct downstream of hsa-miR-375, and its level was negatively mediated by hsa-miR-375. In addition, TLR4 level in Caco-2 cells was upregulated after LPS induction, while hsa-miR-375 level was unchangeable. Knockdown of hsa-miR-375 upregulated NF-κB and pro-inflammatory factors TNF-α, IL-1β, IL-6 and IL-8, and downregulated ZO-1, occludin and anti-inflammatory factor IL-10. Hsa-miR-375 is involved in the pathogenesis of IBD by upregulating TLR4 and inducing NF-κB activation.


Focal organizing pneumonia mimicking lung cancer: a surgeon's view.
Focal organizing pneumonia is a unique form of organizing pneumonia. Little is known regarding its clinical and radiological feature, diagnosis, management, and outcome. Twenty patients with focal organizing pneumonia were investigated and compared with 40 patients with bronchogenic carcinoma. There were 38 men (63.3%) and 22 women (36.7%). The mean age was 55 ± 9.9 years. No specific feature in clinical and radiological manifestation was found to distinguish between focal organizing pneumonia and bronchogenic carcinoma. In patients with focal organizing pneumonia, wedge resection was performed in 12 cases and lobectomy in eight cases. Follow-up was complete with a median period of 26 months (range, 6 to 104 months). All patients were free from recurrence of organizing pneumonia. Clinical and radiologic findings of focal organizing pneumonia are nonspecific, and this unique form of organizing pneumonia is difficult to differentiate from lung cancer. Surgical resection allows both diagnosis and cure. However, considering the benign nature of this disease, major pulmonary resections should be avoided.


Endplate fracture associated with intradiscal dextrose injection.
Intradiscal injections have been used for many years in the treatment of chronic pain. These injections consisted of many substances providing variable pain relief from discogenic pain. Following an intradiscal dextrose injection series, a physically active patient developed superior and inferior lumbar endplate fractures, producing a drastic increase in pain. The pathology of endplate fractures and their contribution to chronic low back pain is subsequently reviewed. A discussion of various intradiscal agent injections is included. Also, the pathophysiology of the production of endplate fractures as related to intradiscal injections is discussed.


Gold-catalyzed tandem reactions of methylenecyclopropanes and vinylidenecyclopropanes.
Gold catalysis is often the key step in the synthesis of natural products, and is a powerful tool for tandem or domino reaction processes. Both gold salts and complexes are among the most powerful soft Lewis acids for electrophilic activation of carbon-carbon multiple bonds toward a variety of nucleophiles. The core of these reactions relies on the interaction between gold catalysts and π-bonds of alkenes, alkynes, and allenes. Activation of functional groups by gold complexes provides a useful and important method for facilitating many different organic transformations with high atom efficiency. Although they are highly strained, methylenecyclopropanes (MCPs) and vinylidenecyclopropanes (VDCPs) are readily accessible molecules that have served as useful building blocks in organic synthesis. Because of their unique structural and electronic properties, significant developments have been made in the presence of transition metal catalysts such as nickel, rhodium, palladium, and ruthenium during the past decades. However, less attention has been paid to the gold-catalyzed chemistry of MCPs and VDCPs. In this Account, we describe gold-catalyzed chemical transformations of MCPs and VDCPs developed both in our laboratory and by other researchers. Chemists have demonstrated that MCPs and VDCPs have amphiphilic properties. When MCPs or VDCPs are activated by a gold catalyst, subsequent nucleophilic attack by other reagents or ring-opening (ring-expansion) of the cyclopropane moiety will occur. However, the C-C double bonds of MCPs and VDCPs can also serve as nucleophilic reagents while more electrophilic reagents are present and activated by gold catalyst, and then further cascade reactions take place as triggered by the release of ring strain of cyclopropane. Based on this strategy, both our group and others have found some interesting gold-catalyzed transformations in recent years. These transformations of MCPs and VDCPs can produce a variety of polycyclic and heterocyclic structures, containing different sized skeletons. Moreover, we have carried out some isotopic labeling experiments and computational studies for mechanistic investigation. These reactions always give the desired products with high level control of chemo-, regio-, and diastereoselectivities, making them highly valuable for the synthesis of natural products and to the pharmaceutical industry and medicine in general.


Hepatic exposure of metformin in patients with non-alcoholic fatty liver disease.
Metformin is first-line treatment of type 2 diabetes mellitus and reduces cardiovascular events in patients with insulin resistance and type 2 diabetes. Target tissue for metformin action is thought to be the liver, where metformin distribution depends on facilitated transport by polyspecific transmembrane organic cation transporters (OCTs). Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease in the western world with strong associations to insulin resistance and the metabolic syndrome, but whether NAFLD affects metformin biodistribution to the liver is not known. In this study, the primary aim was to investigate in vivo hepatic uptake of metformin dynamically in humans with variable degrees of liver affection. As a secondary aim, we wished to correlate hepatic metformin distribution with OCT gene transcription determined in diagnostic liver biopsies. Eighteen patients with biopsy-proven NAFLD were investigated using 11C-metformin PET/CT technique. Gene transcripts of OCTs were determined by real-time polymerase chain reaction (PCR). We observed similar hepatic volume of distribution of metformin between patients with simple steatosis and non-alcoholic steatohepatitis (NASH) (Vd 2.38 ± 0.56 vs. 2.10 ± 0.39, P = 0.3). There was no association between hepatic exposure to metformin and the degree of inflammation or fibrosis, and no clear correlation between metformin distribution and OCT gene transcription. Metformin is distributed to the liver in patients with NAFLD and the distribution is not impaired by inflammation or fibrosis. The findings imply that metformin action in liver in patients with NAFLD may be preserved.


Filtration-assisted exchange transfusion using alpha alpha Hb, an erythrocyte substitute.
A method for improving the efficiency of exchange transfusion to evaluate hemoglobin- (Hb) based erythrocyte substitutes is described. The method uses a continuous-flow hollow-fiber plasma separation filter to remove the erythrocytes while returning 75% of the plasma. The removed volume was replaced with a 14-g/dl solution of human Hb cross-linked between the alpha-chains with bis(3,5-dibromosalicyl)fumarate (alpha alpha Hb). Filtration of 2.76 blood vol in anesthetized swine resulted in a 95% reduction of hematocrit and produced a plasma Hb concentration of 7.63 g/dl. Hyperoncotic Hb solutions cause volume expansion, which reduces the efficiency of exchange but provides hemodynamic stability in the face of decreasing blood viscosity and subsequent intravascular volume loss with Hb redistribution. Filtration-assisted exchange transfusion is rapid, conserves valuable modified Hb, and ensures continuous adequate oxygen delivery.


Constitutive and inducible nitric oxide synthase: role in angiogenesis.
Since the initial report of nitric oxide (NO) activity, enormous progress has been made over the last two decades in the field of NO research. Whereas most physiological responses triggered by moderate concentrations of NO are mediated by soluble guanylate cyclase activation and the subsequent production of cyclic GMP as the major signaling messenger, recent studies have provided evidence of alternative signaling pathways triggered by high concentrations of NO. These signals operate in part through redox-sensitive regulation of transcription factors, gene expression, transcription, cellular activation, proliferation, and cell death. Numerous results converge to indicate a role for NO in physiological and pathological angiogenesis. Experimental data indicate that NO synthase, depending on the isoforms, the timing, and the degree of activation, may display contradictory effects, expressed during both physiological and pathological angiogenesis. The dual personality of NO will be reviewed in the context of the angiogenesis process.


Familiarity modulated synapses model visual cortical circuit novelty responses.
Since environments are constantly in flux, the brain's ability to identify novel stimuli that fall outside its own internal representation of the world is crucial for an organism's survival. Within the mammalian neocortex, inhibitory microcircuits are proposed to regulate activity in an experience-dependent manner and different inhibitory neuron subtypes exhibit distinct novelty responses. Discerning the function of diverse neural circuits and their modulation by experience can be daunting unless one has a biologically plausible mechanism to detect and learn from novel experiences that is both understandable and flexible. Here we introduce a learning mechanism, familiarity modulated synapses (FMSs), through which a network response that encodes novelty emerges from unsupervised synaptic modifications depending only on the presynaptic or both the pre- and postsynaptic activity. FMSs stand apart from other familiarity mechanisms in their simplicity: they operate under continual learning, do not require specialized architecture, and can distinguish novelty rapidly without requiring feedback. Implementing FMSs within a model of a visual cortical circuit that includes multiple inhibitory populations, we simultaneously reproduce three distinct novelty effects recently observed in experimental data from visual cortical circuits in mice: absolute, contextual, and omission novelty. Additionally, our model results in a set of diverse physiological responses across cell subpopulations, allowing us to analyze how their connectivity and synaptic dynamics influences their distinct behavior, leading to predictions that can be tested in experiment. Altogether, our findings demonstrate how experimentally-constrained cortical circuit structure can give rise to qualitatively distinct novelty responses using simple plasticity mechanisms. The flexibility of FMSs opens the door to computationally and theoretically investigating how distinct synapse modulations can lead to a variety of experience-dependent responses in a simple, understandable, and biologically plausible setup.


Risk factors and preventive strategies for breast cancer.
Several risk factors for breast cancer are readily identifiable, but most are not preventable. Hence the importance of screening, as early diagnosis and treatment offer the best hope of saving lives.


Using ecological variables to predict Ross River virus disease incidence in South Australia.
Ross River virus (RRV) disease is Australia's most widespread vector-borne disease causing significant public health concern. The aim of this study was to identify the ecological covariates of RRV risk and to develop epidemic forecasting models in a disease hotspot region of South Australia. Seasonal autoregressive integrated moving average models were used to predict the incidence of RRV disease in the Riverland region of South Australia, an area known to have a high incidence of the disease. The model was developed using data from January 2000 to December 2012 then validated using disease notification data on reported cases for the following year. Monthly numbers of the mosquito Culex annulirostris (β=0.033, p<0.001) and total rainfall (β=0.263, p=0.002) were significant predictors of RRV transmission in the study region. The forecasted RRV incidence in the predictive model was generally consistent with the actual number of cases in the study area. A predictive model has been shown to be useful in forecasting the occurrence of RRV disease, with increased vector populations and rainfall being important factors associated with transmission. This approach may be useful in a public health context by providing early warning of vector-borne diseases in other settings.


Albedo hydrolysis modelling and digestion with reused effluents in the enzymatic peeling process of grapefruits.
Until now, the optimisation of enzymatic peeling of grapefruit in the reactor has been obtained as the result of the semi-qualitative effects of enzyme activity. This work is an attempt to obtain quantified data. The reuse of enzymes to reduce costs in this process is unprecedented in the literature and is the aim of the present work. The optimal conditions determined for the maximum albedo degradation were a temperature of 40.6 °C and a time of 13.1 min in an enzymatic concentration of 0.067 mL enzymatic preparation per gram of peel in each litre of citrate buffer solution. The decrease in relative enzymatic activities in reused effluents was determined, as was the increase in activity when the enzymes were purified. These increases were 15.5% for polygalacturonase and 15.4% for cellulase activity. Optimal temperature, time and the ratio between peel mass and the enzymatic preparation volume were the best conditions for obtaining good peeling efficiency. The effluents from the enzymatic peeling process of the grapefruit still contain appreciable enzymatic activity after the digestion process. Thus, reusing these effluents while maintaining peeling efficacy and a subsequent recovery of the active enzymes by ultra-filtration of the effluents is the way to improve the efficiency of the process.


[Biological effects of chronic radiation exposure on plant populations].
The findings from long-term field studies on biological effects in plant populations inhabiting radioactively contaminated territories contrast in levels and compositions of dose-forming radionuclides are presented. Plant populations developing under radioactive impact show enhanced frequencies of gene and chromosome mutations, and their reproductive potential is inferior to reference populations. Even relatively low levels of technogenic impact are able to increase genetic diversity and destroy regularities inherent for intact populations. Chronic radiation exposure from a certain level appears to be an ecological factor changing genetic structure of wild populations. Data presented indicate the presence of adaptation processes in plant populations in territories with technogenic impact. Under ecological stress, there are selection processes for resistance improvement in plant populations. But an appearance and rate of this process can essentially differ in dependence on radioecological conditions.


Analysis of promoter regions for the spinach chloroplast rbcL, atpB and psbA genes.
A promoter-deletion derivative of the spinach trnM2 gene was used for the identification and characterization of the promoter regions for the spinach chloroplast RuBisCo large subunit (rbcL), ATPase beta-subunit (atpB) and QB-polypeptide (psbA) genes. The DNA sequences 5' upstream from the transcriptional start sites of these genes share homology with the ctp1 and ctp2 arrangement found for the trnM2 transcription unit and the canonical Escherichia coli '-10' and '-35' promoter regions. Synthetic DNA fragments of approximately 40-bp regions, including the defined transcriptional start sites and proximal residues, from rbcL, atpB and psbA, were fused to the trnM2 deletion mutant 51. The promoter-fusion constructs direct the correct transcription of tRNAMet2 in the chloroplast extract with distinct efficiencies. The ctp1- and ctp2-like elements in the trnM2, rbcL and psbA promoter regions can be interchanged to yield functional chimeric promoters of varying strengths. As a result, ctp1 sequences from atpB and psbA, trnM2 and rbcL, respectively, can be ordered TTGACA greater than TTGCTT greater than TTGCGC with respect to their intrinsic strengths. Single base pair changes were introduced into the ctp2-like element in the psbA promoter region. In analogy to similar base pair changes which lower promoter efficiency in E. coli, these mutations result in reduced transcription levels in the chloroplast extract. The data are consistent with a prokaryotic model for chloroplast promoter function.


Cardiac arrest during pregnancy.
Cardiac arrest is a rare event during pregnancy. The pregnant population represents a unique subset of cardiac arrest victims. Not only are there unique causes of circulatory collapse during the pregnant state, but the physiological modifications to the maternal physiology during pregnancy require specific modifications to the standard management of the arrest. Lastly, the pregnant victim presents herself with the challenges of a second patient who needs to be considered in the decision-making process.


Rapid sequence intubation of the pediatric patient. Fundamentals of practice.
Rapid-sequence intubation and rapid sequence induction of general anesthesia are synonyms and refer to the technique of choice for tracheal intubation in many pediatric patients in the emergency department. The principles of safe practice and basic standards of care uniformly apply to all clinical situations in which the technique is performed. RSI has two basic technical components: induction of general anesthesia and direct laryngoscopy with tracheal intubation. The technique is a prescribed protocol that can be modified slightly by the clinical circumstances. RSI is designed to rapidly create ideal intubating conditions, attenuate pathophysiologic reflex responses to direct laryngoscopy and tracheal intubation, and reduce the risk for pulmonary aspiration. Optimal performance requires appropriate training and knowledge, technical skill, and sound medical judgment. Medical and airway evaluation, careful patient selection, recognition of the need for consultation or safer alternatives, thorough familiarity with appropriate drug management, and attention to detail are essential for minimizing the risk for adverse complications. RSI with a rapid injection of preselected dosages of an anesthetic induction agent and muscle relaxant is the pharmacologic technique of choice. Premedication should not be routinely used. Anticipation, recognition, and management of complications are inherent to the competent delivery of all medical care. The unanticipated difficult airway is arguably the most severe complication of RSI, and all individuals performing the technique must prepare in advance a specific plan for this scenario. As with all such skills or procedures, a quality assurance program is important to monitor care, and individuals practicing RSI need to take appropriate steps to maintain competence.


Differential inhibition of 17alpha-hydroxylase and 17,20-lyase activities by three novel missense CYP17 mutations identified in patients with P450c17 deficiency.
The microsomal enzyme cytochrome P450c17 is an important regulator of steroidogenesis. The enzyme has two functions: 17alpha-hydroxylase and 17,20-lyase activities. These functions determine the ability of adrenal glands and gonads to synthesize 17alpha-hydroxylated glucocorticoids (17alpha-hydroxylase activity) and/or sex steroids (17,20-lyase activity). Both enzyme functions depend on correct steroid binding, but it was recently shown that isolated lyase deficiency can also be caused by mutations located in the redox partner interaction domain. In this article we present the clinical history and molecular analysis of two patients with combined 17alpha-hydroxylase/17,20-lyase deficiency and four patients with isolated 17,20-lyase deficiency. In these six patients, four missense CYP17 mutations were identified. Two mutations were located in the steroid-binding domain (F114V and D116V), and the other two mutations were found in the redox partner interaction domain (R347C and R347H). We investigated the activity of these mutated proteins by transfection experiments in COS-1 cells using pregnenolone, progesterone, or their hydroxylated products as a substrate and measuring 17alpha-hydroxylase- and 17,20-lyase-dependent metabolites in the medium. The mutations in the steroid-binding domain (F114V and D116V) of P450c17 caused combined, complete (F114V), or partial (D116V) 17alpha-hydroxylase and 17,20-lyase deficiencies, whereas mutations in the redox partner interaction domain (R347C and R347H) displayed less severe 17alpha-hydroxylase deficiency, but complete 17,20-lyase deficiency. These findings are consistent with the clinical data and support the observation that the redox partner interaction domain is essential for normal 17,20-lyase function of P450c17.


Surgery-related risk factors.
The surgical procedure remains the key element in the multidisciplinary treatment of a wide variety of degenerative, traumatic, tumorous, congenital, and vascular diseases, resulting in an estimated 234 million surgical interventions worldwide each year. Undesired effects are inherent in any medical intervention, but are of particular interest in an invasive procedure for both the patient and the responsible physician. Major topics in current complication research include perception of key factors responsible for complication development, prediction, and whenever possible, prevention of complications. For many years, the technical aspects of surgery and the skills of the surgeon her/himself were evaluated and considered as the main sources of surgical complications. However, recent studies identified many nontechnical perspectives, which could improve the overall quality of surgical interventions. This article reviews selected, recently published data in this field and aims to point out the complexity and multidimensional facets of surgery-related risk factors.


Cyclodextrin based nanosponges for pharmaceutical use: a review.
Nanosponges are a novel class of hyper-crosslinked polymer based colloidal structures consisting of solid nanoparticles with colloidal sizes and nanosized cavities. These nano-sized colloidal carriers have been recently developed and proposed for drug delivery, since their use can solubilize poorly water-soluble drugs and provide prolonged release as well as improve a drug's bioavailability by modifying the pharmacokinetic parameters of actives. Development of nanosponges as drug delivery systems, with special reference to cyclodextrin based nanosponges, is presented in this article. In the current review, attempts have been made to illustrate the features of cyclodextrin based nanosponges and their applications in pharmaceutical formulations. Special emphasis has been placed on discussing the methods of preparation, characterization techniques and applications of these novel drug delivery carriers for therapeutic purposes. Nanosponges can be referred to as solid porous particles having a capacity to load drugs and other actives into their nanocavity; they can be formulated as oral, parenteral, topical or inhalation dosage forms. Nanosponges offer high drug loading compared to other nanocarriers and are thus suitable for solving issues related to stability, solubility and delayed release of actives. Controlled release of the loaded actives and solubility enhancement of poorly water-soluble drugs are major advantages of nanosponge drug delivery systems.


Are there neurologic correlates of in utero cocaine exposure at age 6 years?
At age 6 years, a cohort of non-asphyxiated children, 52 with gestational cocaine exposure and 63 control subjects, underwent a neurologic examination. Groups did not differ on any aspect of the examination. This finding, while reassuring, does not exclude the possibility of more subtle deficits.


Small bowel segment reversal induces intestinal hyperplasia but reduces whole-body growth in massive bowel resected rats.
Surgical reversal of a small bowel segment has been proposed as a means to improve nutritional status in individuals with extensive bowel resection. However, clinical experience remains controversial. The aim of this study was to determine the effects of bowel segment reversal on intestinal adaptation and whole-body anabolism. Male Wistar rats underwent a 70% small bowel resection with (REV) or without (CON) reversal of a 5-cm small bowel segment (5 cm distal to resected segment), or sham-operation (SHAM). After surgery (day 0), rats were fed with powdered diet from day 2 to day 12. Body weight, nitrogen balance, carcass compositions, and serum concentrations of albumin and insulin-like growth factor (IGF)-1 were determined to assess whole-body anabolism. The composition and architecture of the small intestine were measured to assess the intestinal growth response. Serum concentrations of tumor-necrosis factor (TNF)-alpha and interleukin (IL)-6 were measured to assess the response of postoperative cytokines. Surgical reversal significantly increased the intestinal protein and DNA contents in the proximal segment compared with surgical resection. REV rats had a significantly slower rate of weight gain and lower serum levels of albumin and IGF-1, and had significantly greater levels of circulating white blood cells, blood urea nitrogen, and IL-6 compared with CON and SHAM rats. There were no significant differences in serum concentrations of TNF-alpha and carcass percentages of water, protein, and fat among groups. Small bowel segment reversal stimulates jejunal hyperplasia but the surgical reversal induced-elevation in serum IL-6 may eliminate the whole-body anabolism in massive bowel-resected rats.


Sporadic Triple A (Allgrove) Syndrome with Novel Tandem Mutations.
In triple A (Allgrove) syndrome, motor neuron disease is a co-morbid condition. We herein report a 38-year-old Japanese man with triple A (Allgrove) syndrome and novel tandem mutations: a novel c.881delT deletion mutation and c.835C>T localized to the AAAS gene. A nerve conduction study revealed marked axonal damage in several motor nerves. Tandem mutations in the AAAS gene may be involved in co-morbid motor neuron disease and aberrant electrophysiological findings.


In vitro ciprofloxacin resistance profiles among gram-negative bacteria isolated from clinical specimens in a teaching hospital.
To evaluate the trends of ciprofloxacin resistance pattern in commonly isolated gram-negative bacteria over time in a Saudi Arabian teaching hospital. A retrospective analysis was carried out for ciprofloxacin resistance patterns of 10089 isolates of gram-negative bacteria isolated from various clinical specimens submitted to microbiology laboratories at King Fahd Hospital of the University (KFHU), Alkhobar, Saudi Arabia during the period January 2002 to August 2005. Increase in ciprofloxacin resistance rates in these isolates during the years 2002 and 2005 were as follows: Escherichia coli, 23.85 to 33.1%; Klebsiella pneumoniae, 15.93 to 27.55%; Enterobacter cloacae, 12.32 to 18.05%; Enterobacter aerogenes, 4.16 to 25.64%; Pseudomonas aeruginosa 17.05 to 39.53%, Pseudomonas spp., 20.58 to 58.77%; H. influenzae, 00.00% to 11.11% and H. aegyptius, 00.00% to 1.73%. There was a gradual increase in resistance from year 2002 to 2005 in most of the gram negative isolates i.e. E. coli, K. pneumoniae, E. cloacae, E. aerogenes, P. aeruginosa and Pseudomonas spp showing that ciprofloxacin is no more a drug of choice for gram negative infections by these microbes.


Phytochemical and antiulcerogenic properties of rhizomes from Simaba ferruginea St. Hill. (Simaroubaceae).
Simaba ferruginea (Simaroubaceae) is a Brazilian medicinal plant used in traditional medicine to treat several ailments, including gastric ulcers, fever, diarrhea, and dolorous and inflammatory processes. This study examines the chemical composition and antiulcerogenic effects of rhizomes from this plant. Bioassay-guided fractionation led to the isolation of two bioactive indole alkaloids called canthin-6-one (1) and 4-methoxycanthin-6-one (2). The alkaloid fraction and both alkaloids demonstrated potent antiulcerogenic effects when evaluated in gastric lesion-induced animals, as well as significant antinociceptive activity in mice. These results confirm and justify the popular use of S. ferruginea against gastric ulcers and dolorous processes.


Selectivity of antagonists for the Cys-loop native receptors for ACh, 5-HT and GABA in guinea-pig myenteric neurons.
The three most common Cys-loop receptors expressed by myenteric neurons are nACh, 5-HT3 and GABAA . To investigate the function of these proteins researchers have used channel inhibitors such as hexamethonium (antagonist of nACh receptors), ondansetron (antagonist of 5-HT3 receptors), picrotoxin and bicuculline (both antagonists of GABAA receptors). The aim of this study was to investigate the specificity of these inhibitors on Cys-loop receptors of primary cultured neurons obtained from the guinea-pig small intestine. The whole-cell configuration of the patch clamp techniques was used to record membrane currents induced by ACh (IACh ), 5-HT (I5-HT ) and GABA (IGABA ) in the absence and the presence of various concentrations of hexamethonium, ondansetron, picrotoxin or bicuculline. The three Cys-loop receptors present in enteric neurons are expressed independently and they do not cross-desensitized. Hexamethonium inhibited IACh without affecting I5-HT and IGABA . Ondansetron inhibited I5-HT and also IACh but did not affect IGABA . Picrotoxin and bicuculline inhibited I5-HT , IACh and IGABA with different potency, being the lowest potency on 5-HT3 receptors. All these inhibitory effects were concentration dependent and reversible. Our observations showed that except for hexamethonium, all other inhibitors used here show different degrees of selectivity, which has to be considered when these antagonists are used in experimental studies aimed to investigate the functions of these receptors. In particular, in tissues expressing nACh receptors because these are the targets of all other inhibitors used here. The low potency of picrotoxin and bicuculline to inhibit 5-HT3 receptors suggests that these receptors are heteromeric proteins.


A new instrument for kinetics and branching ratio studies of gas phase collisional processes at very low temperatures.
A new instrument dedicated to the kinetic study of low-temperature gas phase neutral-neutral reactions, including clustering processes, is presented. It combines a supersonic flow reactor with vacuum ultra-violet synchrotron photoionization time-of-flight mass spectrometry. A photoion-photoelectron coincidence detection scheme has been adopted to optimize the particle counting efficiency. The characteristics of the instrument are detailed along with its capabilities illustrated through a few results obtained at low temperatures (<100 K) including a photoionization spectrum of n-butane, the detection of formic acid dimer formation, and the observation of diacetylene molecules formed by the reaction between the C2H radical and C2H2.


Genomic evidence for recurrent genetic admixture during the domestication of Mediterranean olive trees (Olea europaea L.).
Olive tree (Olea europaea L. subsp. europaea, Oleaceae) has been the most emblematic perennial crop for Mediterranean countries since its domestication around 6000 years ago in the Levant. Two taxonomic varieties are currently recognized: cultivated (var. europaea) and wild (var. sylvestris) trees. However, it remains unclear whether olive cultivars derive from a single initial domestication event followed by secondary diversification, or whether cultivated lineages are the result of more than a single, independent primary domestication event. To shed light into the recent evolution and domestication of the olive tree, here we analyze a group of newly sequenced and available genomes using a phylogenomics and population genomics framework. We improved the assembly and annotation of the reference genome, newly sequenced the genomes of twelve individuals: ten var. europaea, one var. sylvestris, and one outgroup taxon (subsp. cuspidata)-and assembled a dataset comprising whole genome data from 46 var. europaea and 10 var. sylvestris. Phylogenomic and population structure analyses support a continuous process of olive tree domestication, involving a major domestication event, followed by recurrent independent genetic admixture events with wild populations across the Mediterranean Basin. Cultivated olives exhibit only slightly lower levels of genetic diversity than wild forms, which can be partially explained by the occurrence of a mild population bottleneck 3000-14,000 years ago during the primary domestication period, followed by recurrent introgression from wild populations. Genes associated with stress response and developmental processes were positively selected in cultivars, but we did not find evidence that genes involved in fruit size or oil content were under positive selection. This suggests that complex selective processes other than directional selection of a few genes are in place. Altogether, our results suggest that a primary domestication area in the eastern Mediterranean basin was followed by numerous secondary events across most countries of southern Europe and northern Africa, often involving genetic admixture with genetically rich wild populations, particularly from the western Mediterranean Basin.


Prevention of recurrence of atypical hemolytic uremic syndrome post renal transplant with the use of higher-dose eculizumab .
Eculizumab, a terminal complement inhibitor, has recently been used successfully to both prevent and treat the recurrence of atypical hemolytic uremic syndrome (aHUS) post renal transplantation. We describe a case that highlights the need to monitor the effects of eculizumab on the complement system and in this case alter the dosage. Despite taking the standard recommended dose of eculizumab for an adult, this aHUS patient developed a low-grade thrombotic microangiopathy on biopsy within months of renal transplantation. Complement assays (trough CH50) showed small amounts of residual terminal pathway activity suggesting inadequate complement blockade on 1,200 mg eculizumab every 2 weeks. Following an increase in the dose of eculizumab to 1,500 mg every 2 weeks, lactate dehydrogenase (LDH), proteinuria, and creatinine decreased, and CH50 assay showed 0%. This case emphasizes the need to monitor clinical parameters and complement activity to ensure that adequate therapeutic blockade is achieved. .


Effect of Human Plasma on Hepatic Uptake of Organic Anion-Transporting Polypeptide 1B Substrates: Studies Using Transfected Cells and Primary Human Hepatocytes.
Current challenges with the in vitro-in vivo extrapolation (IVIVE) of hepatic uptake clearance involving organic anion-transporting polypeptide (OATP) 1B1/1B3 hinder drug design strategies. Here we evaluated the effect of 100% human plasma on the uptake clearance using transfected human embryonic kidney (HEK) 293 cells and primary human hepatocytes and assessed IVIVE. Apparent unbound uptake clearance (PSinf,u) increased significantly (P < 0.05) in the presence of plasma (vs. buffer incubations) for about 50% of compounds in both OATP1B1-transfected and wild-type HEK cells. Thus, plasma showed a minimal effect on the uptake ratios. With cultured human hepatocytes, plasma significantly (P < 0.05) increased PSinf,u for 11 of 19 OATP1B substrates (median change of 2.1-fold). Cell accumulation in HEK cells and hepatocytes was also increased for tolbutamide, which is not an OATP substrate. Plasma-to-buffer ratio of PSinf,u obtained in hepatocytes showed a good correlation with unbound fraction in plasma, and the relationship was best described by a "facilitated-dissociation" model. IVIVE was evaluated for the 19 OATP1B substrates using hepatocyte data in the presence of buffer and plasma. PSinf,u from buffer incubations markedly underpredicted hepatic intrinsic clearance (calculated via well stirred and parallel tube models) with an estimated bias of 0.10-0.13. Predictions improved when using PSinf,u from plasma incubations; however, considerable systemic underprediction was still apparent (0.19-0.26 bias). Plasma data with a global scaling factor of 3.8-5.3 showed good prediction accuracy (95% predictions within 3-fold; average fold error = 1.7, bias = 1). In summary, this study offers insight into the effect of plasma on the uptake clearance and its scope in improving IVIVE. SIGNIFICANCE STATEMENT: Our study using diverse anionic compounds shows that human plasma facilitates organic anion-transporting polypeptide 1B-mediated as well as passive uptake clearance, particularly for the highly bound compounds. Leveraging data from transfected human embryonic kidney 293 cells and primary human hepatocytes, we further evaluated mechanisms involved in the observed plasma-facilitated uptake transport. Enhanced hepatic uptake rate in the presence of plasma could be of relevance, as such mechanisms likely prevail in vivo and emphasize the need to maintain physiologically relevant assay conditions to achieve improved translation of transport data.


Pathogenesis and management of the dyslipidemia of the metabolic syndrome.
The metabolic syndrome is a multiplex risk factor for atherosclerotic cardiovascular disease. The core components of the dyslipidemia of the metabolic syndrome, which most likely initiate atherosclerosis, are the "lipid triad" of high plasma triglycerides, low levels of high-density lipoprotein cholesterol (HDL-C), and a preponderance of small, dense low-density lipoprotein (LDL) particles. Postprandial lipemia is also recognized as an important component. Dysregulation of fat metabolism in the liver with overproduction of very-low-density lipoprotein 1 (VLDL1) particles is considered the hallmark of the dyslipidemia of the metabolic syndrome. The cornerstone of therapy is therapeutic lifestyle change. If this does not correct the dyslipidemia, drug therapy may be required. LDL-C lowering with statins is the first-line treatment; however, there is ongoing debate as to whether further aggressive LDL-lowering therapy with higher-dose statins as opposed to comprehensive lipid management is the optimal therapy to reduce cardiovascular risk in subjects with the metabolic syndrome. Combination of a statin with a fibrate and/or nicotinic acid can improve all components of the dyslipidemia. However, there are, as yet, no large cardiovascular outcome studies that show a reduction in cardiovascular morbidity or mortality using combination therapy. The current LDL-C goal in high-risk patients with the metabolic syndrome is below 100 mg/dL, with a non-HDL goal of below 130 mg/dL. However, in very high-risk patients, such as those with established cardiovascular disease, an LDL-C goal of below 70 mg/dL is recommended.


LPS-induced migration of peritoneal B-1 cells is associated with upregulation of CXCR4 and increased migratory sensitivity to CXCL12.
B-1 cells, which constitute a predominant lymphocyte subset in serosal cavities and produce most of natural antibodies, are subdivided into the CD5(+) B-1a and CD5(-) B-1b cell subpopulations, but the differential roles of B-1a and B-1b cells are not well understood. We report that B-1a cells preferentially migrate out of the peritoneal cavity and upregulate the expression of CXCR4 with heightened sensitivity to CXCL12 and CXCL13 upon LPS treatment compared to B-1b and B-2 cells. Whereas B-1a cells were slightly more abundant than B-1b and B-2 cells in the homeostatic condition, the number of B-1a cells preferentially decreased 48 hr after LPS treatment. The decrease in the peritoneal B-1a cell number was accompanied with increased migration of B-1a cells toward CXCL-12 and CXCL-13 in in vitro transmigration assay using peritoneal B cells from LPS treated mice. The expression level of CXCR4, but not of CXCR5, was also more prominently increased in B-1a cells upon LPS stimulation. LPS-stimulated B-1a cells did not accumulate in omental milky spots in contrast to B-2 cells. These results suggest that B-1a cells actively migrate out of the peritoneal cavity through the regulation of the migratory responsiveness to chemokines and actively participate in systemic immune responses.


Effect of jet injection on infectivity of measles, mumps, and rubella vaccine in a bench model.
Disposable-syringe jet injectors (DSJIs) with single-use, auto disable, needle-free syringes offer the opportunity to avoid hazards associated with injection using a needle and syringe. Clinical studies have evaluated DSJIs for vaccine delivery, but most studies have focused on inactivated, subunit, or DNA vaccines. Questions have been raised about possible damage to live attenuated viral vaccines by forces generated during the jet injection process. This study examines the effect of jet injection on the integrity of measles, mumps, and rubella vaccine (MMR), measured by viral RNA content and infectivity. Three models of DSJIs were evaluated, each generating a different ejection force. Following jet injection, the RNA content for each of the vaccine components was measured using RT-qPCR immediately after injection and following passage in Vero cells. Jet injection was performed with and without pig skin as a simulation of human skin. There was little to no reduction of RNA content immediately following jet injection with any of the three DSJIs. Samples passaged in Vero cells showed no loss in infectivity of the measles vaccine following jet injection. Mumps vaccine consistently showed increased replication following jet injection. Rubella vaccine showed no loss after jet injection alone but some infectivity loss following injection through pig skin with two of the devices. Overall, these data demonstrated that forces exerted on a live attenuated MMR vaccine did not compromise vaccine infectivity. The bench model and protocol used in this study can be applied to evaluate the impact of jet injection on other live virus vaccines.


An integrated prognostic model for diffuse large B-cell lymphoma treated with immunochemotherapy.
Diffuse large B-cell lymphoma (DLBCL), the most frequent non-Hodgkin's lymphoma subtype, is characterized by strong biological, morphological, and clinical heterogeneity, but patients are treated with immunochemotherapy in a relatively homogeneous way. Here, we have used a customized NanoString platform to analyze a series of 197 homogeneously treated DLBCL cases. The platform includes the most relevant genes or signatures known to be useful for predicting response to R-CHOP (Rituximab, Cyclophosphamide, Doxorubicin, Vincristine, and Prednisone) in DLBCL cases. We generated a risk score that combines the International Prognostic Index with cell of origin and double expression of MYC/BCL2, and stratified the series into three groups, yielding hazard ratios from 0.15 to 5.49 for overall survival, and from 0.17 to 5.04 for progression-free survival. Group differences were highly significant (p < 0.0001), and the scoring system was applicable to younger patients (<60 years of age) and patients with advanced or localized stages of the disease. Results were validated in an independent dataset from 166 DLBCL patients treated in two distinct clinical trials. This risk score combines clinical and biological data in a model that can be used to integrate biological variables into the prognostic models for DLBCL cases.


A systematic review of interventions promoting clinical information retrieval technology (CIRT) adoption by healthcare professionals.
This paper presents the evidence on the effectiveness of interventions promoting the use of clinical information retrieval technologies (CIRTs) by healthcare professionals. We electronically searched articles published between January 1990 and March 2008 using following inclusion criteria: (1) participants were healthcare professionals; (2) specific intervention promoted CIRT adoption; (3) studies were randomised controlled trials, controlled clinical trials, controlled before and after studies or interrupted time series analyses; and (4) they objectively reporting measured outcomes on CIRT use. We found nine studies focusing on CIRT use. Main outcomes measured were searching skills and/or frequency of use of electronic databases by healthcare professionals. Three studies reported a positive effect of the intervention on CIRT use, one showed a positive impact post-intervention, and four studies failed to demonstrate significant intervention effect. The ninth study examined financial disincentives, and found a significant negative effect of introducing user fees for searching MEDLINE in clinical settings. A meta-analysis showed that educational meetings were the only type of interventions reporting consistent positive effects on CIRT adoption. CIRT is an information and communication technology commonly used in healthcare settings. Interventions promoting CIRT adoption by healthcare professionals have shown some success in improving searching skills and use of electronic databases. However, the effectiveness of these interventions remains uncertain and more rigorous studies are needed.


[A Case of Parkinson's Disease Associated with Repeated Deterioration of Akinesia/Bradykinesia Due to Hypothermia].
In 2012, a 69-year-old man noticed slowness in his movements and was diagnosed with Parkinson's disease (PD). In December 2017, he was admitted to a hospital with a diagnosis of hypothermia. One month later, in January 2018, he had difficulty moving around at home and was admitted to our hospital because of impaired consciousness. On admission, his consciousness was rated as level II-10, his body temperature was 32.8℃, and he was pale and had extrapyramidal symptoms. Electrocardiogram showed T-wave flattening and the first degree of atrioventricular block. Warming was started with improvement of the level of consciousness on the sixth hospital day. His cognitive function was normal (HDS-R 27/30), but the Schellong test (an orthostatic blood pressure test) showed a marked decrease in systolic blood pressure upon standing. Although sympathetic skin and flow responses were almost normal, except for low levels of basal skin blood flow, we speculated that the patient had impaired thermoregulation due to central autonomic dysfunction. In the literature, such impaired thermoregulation was suggested as the cause of hypothermia in seven patients with PD. Therefore, we recommend that patients with PD and repeated hypothermia should be instructed to maintain a suitable room temperature and to dress warmly. (Received 25 June, 2021; Accepted 31 August, 2021; Published 1 December, 2021).


Methods for Implant Acceptance and Wound Healing: Material Selection and Implant Location Modulate Macrophage and Fibroblast Phenotypes.
This review focuses on materials and methods used to induce phenotypic changes in macrophages and fibroblasts. Herein, we give a brief overview on how changes in macrophages and fibroblasts phenotypes are critical biomarkers for identification of implant acceptance, wound healing effectiveness, and are also essential for evaluating the regenerative capabilities of some hybrid strategies that involve the combination of natural and synthetic materials. The different types of cells present during the host response have been extensively studied for evaluating the reaction to different materials and there are varied material approaches towards fabrication of biocompatible substrates. We discuss how natural and synthetic materials have been used to engineer desirable outcomes in lung, heart, liver, skin, and musculoskeletal implants, and how certain properties such as rigidity, surface shape, and porosity play key roles in the progression of the host response. Several fabrication strategies are discussed to control the phenotype of infiltrating macrophages and fibroblasts: decellularization of scaffolds, surface coatings, implant shape, and pore size apart from biochemical signaling pathways that can inhibit or accelerate unfavorable host responses. It is essential to factor all the different design principles and material fabrication criteria for evaluating the choice of implant materials or regenerative therapeutic strategies.


Toxicokinetics of dothiepin: 2 case reports.
In this article, 2 cases of intentional dothiepin intoxication are presented. Both patients survived after receiving appropriate supportive care. The dothiepin and metabolite levels were measured at different times after ingestion. The initial levels of dothiepin were 1900 microg/L in case 1 and 5500 microg/L in case 2. In case 1, a toxicokinetic model was fitted, suggesting a higher peak level, corresponding with the QRS duration and clinical symptoms. In case 2, a combined dothiepin-ethanol intoxication was seen, complicating the interpretation of clinical parameters, such as the QRS duration. In conclusion, the severity of dothiepin intoxication is poorly predicted by plasma levels alone, as time of ingestion can be critical for interpretation. However, especially in combined intoxications, interpretation of the QRS duration may also fail. Therefore, it is important to evaluate the whole range of clinical parameters, QRS duration, clinical symptoms, dothiepin concentration(s), and other possible intoxications.


Adaptive Aromaticity in 18e Metallapentalenes.
Complexes with aromaticity in both the lowest singlet state (S0) and the lowest triplet state (T1) (denoted as adaptive aromaticity) are rare because according to Hückel's and Baird's rules, a species could be aromatic in either the S0 or T1 state in most cases. Thus, it is particularly challenging to design species with adaptive aromaticity. Previous reports on adaptive aromaticity were mainly focused on 16e metallapentalenes. Here, we demonstrate that 18e metallapentalenes could possess adaptive aromaticity supported by a set of aromaticity indices when the nitrido and imido ligands are introduced via density functional theory calculations. Further investigation suggests that the metal-carbon bond strength plays an important role in the S0 state aromaticity and the T1 state aromaticity could be attributed to spin electron localization. All these findings could be useful for the development of metallaaromatic chemistry.


Positive contribution of macrofaunal biodiversity to secondary production and seagrass carbon metabolism.
Coastal vegetated habitats such as seagrasses are known to play a critical role in carbon cycling and the potential to mitigate climate change, as blue carbon habitats have been repeatedly highlighted. However, little information is known about the role of associated macrofauna communities on the dynamics of critical processes of seagrass carbon metabolism (e.g., respiration, turnover, and production). We conducted a field study across a spatial gradient of seagrass meadows involving variable environmental conditions and macrobenthic diversity to investigate (1) the relationship between macrofauna biodiversity and secondary production (i.e., consumer incorporation of organic matter per time unit), and (2) the role of macrofauna communities in seagrass organic carbon metabolism (i.e., respiration and primary production). We show that, although several environmental factors influence secondary production, macrofauna biodiversity controls the range of local seagrass secondary production. We demonstrate that macrofauna respiration rates were responsible for almost 40% of the overall seafloor community respiration. Macrofauna represented on average >25% of the total benthic organic C stocks, high secondary production that is likely to become available to upper trophic levels of the coastal food web. Our findings support the role of macrofauna biodiversity in maintaining productive ecosystems, implying that biodiversity loss due to ongoing environmental change yields less productive seagrass ecosystems. Therefore, the assessment of carbon dynamics in coastal habitats should include associated macrofauna biodiversity elements if we aim to obtain robust estimates of global carbon budgets required to implement management actions for the sustainable functioning of the world's coasts.


Chum salmon egg extracts induce upregulation of collagen type I and exert antioxidative effects on human dermal fibroblast cultures.
Components of fish roe possess antioxidant and antiaging activities, making them potentially very beneficial natural resources. Here, we investigated chum salmon eggs (CSEs) as a source of active ingredients, including vitamins, unsaturated fatty acids, and proteins. We incubated human dermal fibroblast cultures for 48 hours with high and low concentrations of CSE extracts and analyzed changes in gene expression. Cells treated with CSE extract showed concentration-dependent upregulation of collagen type I genes and of multiple antioxidative genes, including OXR1, TXNRD1, and PRDX family genes. We further conducted in silico phylogenetic footprinting analysis of promoter regions. These results suggested that transcription factors such as acute myeloid leukemia-1a and cyclic adenosine monophosphate response element-binding protein may be involved in the observed upregulation of antioxidative genes. Our results support the idea that CSEs are strong candidate sources of antioxidant materials and cosmeceutically effective ingredients.


Molecular mechanisms for the activation of voltage-independent Ca2+ channels by endothelin-1 in chinese hamster ovary cells stably expressing human endothelin(A) receptors.
We demonstrated recently that in Chinese hamster ovary cells stably expressing human recombinant endothelin(A) receptors (CHO-ET(A)R), endothelin-1 (ET-1) activates two types of Ca2+-permeable nonselective cation channels (designated NSCC-1 and NSCC-2) and a store-operated Ca2+ channel (SOCC), which can be distinguished by Ca(2+) channel blockers such as 1-[beta-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenylethyl]-1H-imidazole hydrochloride (SK&F 96365) and (R,S)-(3,4-dihydro-6,7-dimethoxy-isochinolin-1-yl)-2-phenyl-N,N-di[2-(2,3,4-trimethoxyphenyl)ethyl]acetamid mesylate (LOE 908). We also reported that CHO-ET(A)R couples with G12 in addition to G(q) and G(s). The purpose of the present study was to identify the G proteins involved in the activation of these Ca2+ channels by ET-1, using mutated ET(A)Rs with coupling to either G(q) or G(s)/G12 (designated ET(A)RDelta385 and SerET(A)R, respectively) and a dominant-negative mutant of G12 (G12G228A). ET(A)RDelta385 is truncated immediately downstream of Cys385 in the C terminus as palmitoylation sites, whereas SerET(A)R is unpalmitoylated because of substitution of all the cysteine residues to serine (Cys383Cys385-388 --> Ser383Ser385-388). In CHO-ET(A)RDelta385, stimulation with ET-1 activated only SOCC. In CHO-SerET(A)R or CHO-ET(A)R pretreated with U73122, an inhibitor of phospholipase C (PLC), ET-1 activated only NSCC-1. Dibutyryl cAMP alone did not activate any Ca2+ channels in the resting and ET-1-stimulated CHO-SerET(A)R. Microinjection of G12G228A abolished the activation of NSCC-1 and NSCC-2 in CHO-ET(A)R and that of NSCC-1 in CHO-SerET(A)R. These results indicate that ET(A)R activates three types of Ca2+ channels via different G protein-related pathways. NSCC-1 is activated via a G12-dependent pathway, NSCC-2 via G(q)/PLC- and G12-dependent pathways, and SOCC via a G(q)/PLC-dependent pathway.


A narrative review for the Hippo-YAP pathway in cancer survival and immunity: the Yin-Yang dynamics.
The Hippo-YAP pathway is fast becoming a key instrument in regulating cancers. Binary oppositions, also known as the Yin-Yang dynamics in Chinese, have emerged in its effects. Some oppositions are attributable to in vitro and in vivo experimental conditions, some are due to differences between cancers or species, some are derived from its inherent duality endowed by upstream and downstream signaling, and some are yet unresolved mysteries. However, as bewildering they are, few have been noticed and defined so far. In this review, we first look back on the Hippo pathway which was initially identified more than a decade ago. We then focus on tumor biology, especially some latest popular mechanisms that regulate tumor cell survival, such as ferroptosis, autophagy, and apoptosis. The third chapter is concerned with the findings of the relationship between the Hippo pathway and tumor immunity on the microenvironment in which tumor cells progress. In each of these main sections the contradictory points of the Hippo pathway are elucidated and thoroughly examined. On one hand, the Yin-Yang dynamics of the Hippo pathway brings about considerable challenges for current research; on the other hand, this work will generate fresh insight into it and offer opportunities for subsequent drug development for cancer and regenerative medicine.


Stochastic Effects in Retrotransposon Dynamics Revealed by Modeling under Competition for Cellular Resources.
Transposons are genomic elements that can relocate within a host genome using a 'cut'- or 'copy-and-paste' mechanism. They make up a significant part of many genomes, serve as a driving force for genome evolution, and are linked with Mendelian diseases and cancers. Interactions between two specific retrotransposon types, autonomous (e.g., LINE1/L1) and nonautonomous (e.g., Alu), may lead to fluctuations in the number of these transposons in the genome over multiple cell generations. We developed and examined a simple model of retrotransposon dynamics under conditions where transposon replication machinery competed for cellular resources: namely, free ribosomes and available energy (i.e., ATP molecules). Such competition is likely to occur in stress conditions that a malfunctioning cell may experience as a result of a malignant transformation. The modeling revealed that the number of actively replicating LINE1 and Alu elements in a cell decreases with the increasing competition for resources; however, stochastic effects interfere with this simple trend. We stochastically simulated the transposon dynamics in a cell population and showed that the population splits into pools with drastically different transposon behaviors. The early extinction of active Alu elements resulted in a larger number of LINE1 copies occurring in the first pool, as there was no competition between the two types of transposons in this pool. In the other pool, the competition process remained and the number of L1 copies was kept small. As the level of available resources reached a critical value, both types of dynamics demonstrated an increase in noise levels, and both the period and the amplitude of predator-prey oscillations rose in one of the cell pools. We hypothesized that the presented dynamical effects associated with the impact of the competition for cellular resources inflicted on the dynamics of retrotransposable elements could be used as a characteristic feature to assess a cell state, or to control the transposon activity.


Broad phenotypic spectrum of germ line 7p12.1 microdeletions encompassing the IKZF1 gene includes predisposition to acute lymphoblastic leukemia.
Microdeletions of 7p12.1 encompassing the IKZF1 gene locus are rare, with few cases reported. The common phenotype includes intellectual disability, overgrowth, and facial dysmorphism accompanied, albeit rarely, by congenital anomalies. Haploinsufficiency of IKZF1 predisposes individuals to childhood acute lymphoblastic leukemia (ALL). In this study, we comprehensively analyzed the frequency of 7p12.1 deletions among 4581 Polish individuals who underwent chromosomal microarray testing for unexplained developmental delay, intellectual disability, and/or congenital anomalies. Two unrelated individuals (0.04%) with a de novo interstitial 7p12.1 microdeletion encompassing IKZF1 were identified. One developed ALL. Analysis of the incidence and the phenotype of constitutional 7p12.1 microdeletion, which based on the previously annotated patients data in public databases and literature reports, revealed 21 cases including five patients diagnosed with ALL.