Effects of repeated cocaine exposure and withdrawal on voluntary ethanol drinking, and the expression of glial glutamate transporters in mesocorticolimbic system of P rats.
Glutamatergic neurotransmission within the brain's reward circuits plays a major role in the reinforcing properties of both ethanol and cocaine. Glutamate homeostasis is regulated by several glutamate transporters, including glutamate transporter type 1 (GLT-1), cystine/glutamate transporter (xCT), and glutamate aspartate transporter (GLAST). Cocaine exposure has been shown to induce a dysregulation in glutamate homeostasis and a decrease in the expression of GLT-1 and xCT in the nucleus accumbens (NAc). In this study, alcohol preferring (P) rats were exposed to free-choice of ethanol (15% and 30%) and/or water for five weeks. On Week 6, rats were administered (i.p.) cocaine (10 and 20mg/kg) or saline for 12 consecutive days. This study tested two groups of rats: the first group was euthanized after seven days of repeated cocaine i.p. injection, and the second group was deprived from cocaine for five days and euthanized at Day 5 after cocaine withdrawal. Only repeated cocaine (20mg/kg, i.p.) exposure decreased ethanol intake from Day 3 through Day 8. Co-exposure of cocaine and ethanol decreased the relative mRNA expression and the expression of GLT-1 in the NAc but not in the medial prefrontal cortex (mPFC). Importantly, co-exposure of cocaine and ethanol decreased relative expression of xCT in the NAc but not in the mPFC. Our findings demonstrated that chronic cocaine exposure affects ethanol intake; and ethanol and cocaine co-abuse alters the expression of glial glutamate transporters.


Entamoeba dispar, but not E. histolytica, detected in a colony of chimpanzees in Japan.
Chimpanzees (Pan troglodytes) residing in the Kumamoto Primate Research Park, Sanwa Kagaku Kenkyusho, were surveyed for the presence of intestinal parasites. Stool samples from 107 chimpanzees were examined by microscopy after formalin-ether sedimentation. Of these animals, 100 were infected with at least 1 species of ameba. The positivity rates recorded were as follows: Entamoeba coli, 88%; E. histolytica/E. dispar, 48%; E. hartmanni, 15%; Iodamoeba buetschlii, 8%; Endolimax nana, 4%; and Entamoeba chattoni, 2%. Polymerase chain reaction (PCR) analysis to distinguish between E. histolytica and E. dispar was performed on these samples. E. dispar DNA was detected in 60 of 107 samples (56%), including 9 that had been microscopically determined to be negative for E. histolytica/ E. dispar. In contrast, no E. histolytica DNA was detected in the 107 samples. Zymodeme analysis indicated that 10 isolates were E. dispar. When 104 chimpanzees were examined serologically for E. histolytica infection, 1 sample was scored as positive by indirect hemagglutination and another was found to be positive by an indirect fluorescent antibody test. However, both specimens were borderline-positive and were clearly negative in other tests, suggesting that they might be false-positives. These results demonstrate that the pathogenic E. histolytica was absent in this colony, regardless of the high degree of prevalence of other amebas. For an accurate diagnosis, PCR analysis is recommended in addition to microscopic examination.


Analysis of the transthyretin-like (TTL) gene family in Ostertagia ostertagi--comparison with other strongylid nematodes and Caenorhabditis elegans.
The transthyretin-like (ttl) gene family is one of the largest conserved nematode-specific gene families, coding for a group of proteins with significant sequence similarity to transthyretins (TTR) and transthyretin-related proteins (TRP). In the present study, we investigated the ttl family in Ostertagia ostertagi (a nematode of the abomasum of cattle). Mining of expressed sequence tag (EST) databases revealed the presence of at least 18 ttl genes in O. ostertagi (Oo-ttl), most of which are constitutively transcribed from the free-living, third larval stage onwards. The full-length cDNA of one of these genes (Oo-ttl-1) was amplified and cloned for recombinant expression. Western blot analysis using a specific antiserum showed that the native protein Oo-TTL-1 was highly present in the excretory-secretory (ES) products of adults of O. ostertagi. The protein was immunolocalized to the pseudocoelomic fluid of adult worms. A phylogenetic-bioinformatic analysis of all amino acid sequence data for TTL proteins from a range of strongylid nematodes showed that they could be divided into at least five different classes. This classification was based on conserved amino acids in the first TTL signature domain and the number and location of cysteine residues. The biological role(s) of the TTLs in nematode biology is still unclear. A theoretical three-dimensional model of Oo-TTL-1 indicated that it had a similar structure to TTRs (i.e., containing β-sheets, arranged in a β-sandwich). In contrast to TTRs, competitive binding studies using recombinant Oo-TTL-1 indicated that the protein was devoid of any hydrophobic ligand- or thyroid hormone-binding properties. Finally, combinatorial analysis by double-stranded RNA interference of five ttl genes in the free-living nematode Caenorhabditis elegans did not reveal any visible phenotypes. More information on the transcription profile and tissue distribution of TTLs in nematodes is needed to provide new insights into the biological role of this gene family.


X chromosome evidence for ancient human histories.
Diverse African and non-African samples of the X-linked PDHA1 (pyruvate dehydrogenase E1 alpha subunit) locus revealed a fixed DNA sequence difference between the two sample groups. The age of onset of population subdivision appears to be about 200 thousand years ago. This predates the earliest modern human fossils, suggesting the transformation to modern humans occurred in a subdivided population. The base of the PDHA1 gene tree is relatively ancient, with an estimated age of 1.86 million years, a late Pliocene time associated with early species of Homo. PDHA1 revealed very low variation among non-Africans, but in other respects the data are consistent with reports from other X-linked and autosomal haplotype data sets. Like these other genes, but in conflict with microsatellite and mitochondrial data, PDHA1 does not show evidence of human population expansion.


Wingless and Notch signaling provide cell survival cues and control cell proliferation during wing development.
Tissue growth during animal development depends on the coordination of cell proliferation and cell death. The EGF-receptor/MAPK, Hedgehog, Dpp, Wingless (Wg) and Notch signaling pathways have been implicated in growth control in the developing Drosophila wing. In this report, we examine the effects of Notch and Wg on growth in terms of cell proliferation and cell survival. Reduction of Wg signaling impaired compartment and clonal growth, and increased cell death. Inhibition of apoptosis in cells deficient for Wg signaling only partially rescued the clone growth defect, suggesting that Wg is also required to promote cell proliferation. This is supported by the finding that ectopic expression of Wg caused over-proliferation of cells in the proximal wing. Localized activation of Notch had non-autonomous effects on cell proliferation. However, only part of this effect was attributable to Notch-dependent induction of Wg, suggesting that other Notch-inducible signaling molecules contribute to the control of cell proliferation in the wing.


Strategies for Bacteriophage T5 Mutagenesis: Expanding the Toolbox for Phage Genome Engineering.
Phage genome editing is crucial to uncover the molecular mechanisms of virus infection and to engineer bacteriophages with enhanced antibacterial properties. Phage genetic engineering relies mostly on homologous recombination (HR) assisted by the targeted elimination of wild-type phages by CRISPR-Cas nucleases. These strategies are often less effective in virulent bacteriophages with large genomes. T5 is a virulent phage that infects Escherichia coli. We found that CRISPR-Cas9 system (type II-A) had ununiform efficacies against T5, which impairs a reliable use of CRISPR-Cas-assisted counterselection in the gene editing of T5. Here, we present alternative strategies for the construction of mutants in T5. Bacterial retroelements (retrons) proved to be efficient for T5 gene editing by introducing point mutations in the essential gene A1. We set up a protocol based on dilution-amplification-screening (DAS) of phage pools for mutant enrichment that was used to introduce a conditional mutation in another essential gene (A2), insert a new gene (lacZα), and construct a translational fusion of a late phage gene with a fluorescent protein coding gene (pb10-mCherry). The method should be applicable to other virulent phages that are naturally resistant to CRISPR/Cas nucleases.


Are Doctors the Best People to Manage Gout? Is There a Role for Nurses and Pharmacists?
To discuss alternate models of long-term gout management RECENT FINDINGS: Nurse-led care of gout appears to improve the uptake of and adherence to urate-lowering treatment in a research setting. Less impressive improvements were achieved with pharmacist-led remote management of gout; however, both strategies were more effective than usual primary care provider management of gout. Individualised education about gout, patient involvement in decision-making, and access to trained support in managing side-effects and gout flares can improve the uptake of fine and adherence to urate-lowering treatment. This may be best achieved with nurse-led care of gout. However, further research is required to evaluate if the model of nurse-led care of gout can be implemented in routine clinical practice and in different healthcare systems.


Antibody isotypes to a Paracoccidioides brasiliensis somatic antigen in sub-acute and chronic form paracoccidioidomycosis.
This report describes the differences in isotype antibody reactivity against a crude Paracoccidioides brasiliensis antigenic preparation in the sub-acute (SAF) and chronic (CF) forms of paracoccidioidomycosis before treatment. IgG antibodies were detected in all patients, with a slightly but not significantly higher reactivity in the SAF. IgG1 antibodies were present, frequently at high levels, in both forms, whereas IgG3 was always low or absent. IgG2 antibodies were detectable in most patients, but at high levels in only a few CF patients. IgG4 was found mainly in SAF patients, whereas IgA was detected almost only in CF patients, probably due to a Th2 pattern of immune response in the more severe SAF, and the characteristic mucosal involvement of the CF, respectively. Immunoblot analysis showed that, in addition to the 43-kDa immunodominant fraction, other less well-characterised fractions were also recognised differentially by the isotypes and deserve further investigation.


CYP72A67 Catalyzes a Key Oxidative Step in Medicago truncatula Hemolytic Saponin Biosynthesis.
In the Medicago genus, triterpenic saponins are bioactive secondary metabolites constitutively synthesized in the aerial and subterranean parts of plants via the isoprenoid pathway. Exploitation of saponins as pharmaceutics, agrochemicals and in the food and cosmetic industries has raised interest in identifying the enzymes involved in their synthesis. We have identified a cytochrome P450 (CYP72A67) involved in hemolytic sapogenin biosynthesis by a reverse genetic TILLING approach in a Medicago truncatula ethylmethanesulfonate (EMS) mutagenized collection. Genetic and biochemical analyses, mutant complementation, and expression of the gene in a microsome yeast system showed that CYP72A67 is responsible for hydroxylation at the C-2 position downstream of oleanolic acid synthesis. The affinity of CYP72A67 for substrates with different substitutions at multiple carbon positions was investigated in the same in vitro yeast system, and in relation to two other CYP450s (CYP72A68) responsible for the production of medicagenic acid, the main sapogenin in M. truncatula leaves and roots. Full sib mutant and wild-type plants were compared for their sapogenin profile, expression patterns of the genes involved in sapogenin synthesis, and response to inoculation with Sinorhizobium meliloti. The results obtained allowed us to revise the hemolytic sapogenin pathway in M. truncatula and contribute to highlighting the tissue specificities (leaves/roots) of sapogenin synthesis.


Children's reactions to transgressions: effects of the actor's apology, reputation and remorse.
This experiment examined children's reactions to a transgression in which one child's property was damaged by another who (a) had a reputation as a good or bad child, (b) apologized or did not, and (c) later expressed remorse when talking about the incident or was happy and unremorseful. As expected, actors who had a good reputation or were remorseful were seen as more likable, as having better motives, as doing the damage unintentionally, as more sorry and as less blameworthy. Further, actors who were good and remorseful were punished least, suggesting that punishment was applied in a rehabilitative fashion. The actor's reputation determined how his or her actions were interpreted: bad actors were seen as more worried about punishment when they expressed remorse and older children thought they apoligized merely to avoid punishment. Interestingly, apologies were effective in reducing punishment and making the actor seem more likable, and this was true irrespective of the other factors. The apology-forgiveness script may be such an ingrained aspect of social life that its appearance automatically improves the actor's position. The reactions of second and fifth graders were generally similar, although the younger children displayed less coherent relationships between judgements.


Long-term results of combined chemotherapy-radiotherapy approach in Hodgkin's disease: superiority of ABVD plus radiotherapy versus MOPP plus radiotherapy.
In an attempt to reduce some of the delayed sequelae associated with combined modality therapy in Hodgkin's disease, we randomly tested stages IIB, IIIA, and IIIB MOPP (mechlorethamine, vincristine, procarbazine, and prednisone) v ABVD (Adriamycin, bleomycin, vinblastine, and dacarbazine). In 232 previously untreated patients, three cycles of either combination preceded and followed extensive irradiation. The complete remission rate was 80.7% following MOPP and 92.4% following ABVD (P less than .02). The 7-year results indicated that ABVD was superior to MOPP in terms of freedom from progression (80.8% v 62.8%; P less than .002), relapse-free survival (87.7% v 77.2%; P = .06), and overall survival (77.4% v 67.9%; P = .03). Moreover, the comparative iatrogenic morbidity showed that irreversible gonadal dysfunction as well as acute leukemia occurred only in patients subjected to MOPP, while cardiopulmonary studies failed to document significant laboratory differences between the two treatment groups. Present findings indicate that ABVD followed by extensive irradiation represents a valid therapeutic alternative to the widely used alkylating agent-containing regimens plus radiotherapy.


Pharmacological properties of gamma-aminobutyric acid-, glutamate-, and aspartate-induced depolarizations in cultured astrocytes.
Differentiated glial fibrillary acidic protein-positive astrocytes in homogeneous cultures of early postnatal rat cerebral hemispheres respond by membrane depolarization to gamma-aminobutyric acid (GABA), glutamate, and aspartate with a threshold concentration of approximately 10(-5) M. The GABA-induced depolarization is antagonized by two blockers of the neuronal GABAA receptor, picrotoxin and bicuculline, but is not affected by the uptake blockers beta-alanine or nipecotic acid. An agonist of the GABAA receptor, muscimol, produces a dose-response curve similar to that of GABA, whereas the agonist of the GABAB receptor, baclofen, did not alter the membrane potential. When repetitive pulses of GABA are given to one cell, its responsiveness depends on the time interval between pulses. Within 30 sec after termination of the first pulse the cell remains unresponsive to the second pulse. With increased time intervals between the pulses, reactivity toward GABA recovers. Five minutes after the first pulse the cell regains 75% of its initial depolarization peak. Aspartate results in a depolarization similar in size and time course to that induced by glutamate. The glutamate agonists, quisqualate and ibotenate, and kainate are less potent than glutamate. N-Methyl-D-aspartate has no effect on the membrane potential of astrocytes. The pharmacological features of the glutamate response are therefore similar to those of the receptor mediating neuronal glutamate transport.


Tn1721-encoded tetracycline resistance: mapping of structural and regulatory genes mediating resistance.
The genes encoding inducible tetracycline resistance in Tn1721 were located in a 2.1-kilobase portion of the transposon. Using deletions and insertions, we mapped and characterized two tet genes by their mutant phenotypes. Two tetracycline-inducible polypeptides synthesized in minicells were assigned to the tet genes. The polarity of the tet genes was determined by employing a deletion and a gene fusion which altered the carboxy termini of the polypeptides. The gene responsible for resistance (tetA) encompasses 1,250 base pairs and encodes a membrane-bound protein with an apparent molecular weight of 34,000. The second gene (tetR) encompasses at least 650 base pairs and encodes a soluble 26,000-dalton protein, identified by complementation analysis as the repressor. The two adjacent genes have opposite transcriptional polarity, suggesting that the sites controlling their expression are located in the intercistronic region between tetA and tetR.


Experimental and theoretical study of wide hysteresis cycles in 1550 nm VCSELs under optical injection.
We present experimental results of output power bistability in a vertical-cavity surface-emitting laser under optical injection induced by frequency detuning or power variation of the master laser. An ultra-wide hysteresis cycle of 3.7 nm (473.3 GHz) is achieved through frequency detuning, which is more than 11 times wider than that achieved in the state-of-the-art (37 GHz). Furthermore, the width of injection power induced hysteresis cycle we achieved is as large as 7.3 dB. We theoretically analyzed the hysteresis cycles based on standard optical injection locking rate equations including the interference effect of master laser reflection and found excellent agreement with experimental results.


Demonstration by burst-phase analysis of a robust folding intermediate in the FF domain.
The role of intermediates in the folding reaction of single-domain proteins is a controversial issue. It was previously shown by different methods that an on-pathway intermediate is populated in the presence of sodium sulphate during the folding of the FF domain from HYPA/FBP11. Here we demonstrate using analysis of the amplitudes of kinetic traces that this burst-phase folding intermediate is present at different salt concentration and at various pH, and is also found in roughly 30 site-directed mutants. The intermediate appears robust to changing conditions and thus fulfils an important criterion for a productive molecular species on the folding reaction pathway.


Results from the Periodontitis and Vascular Events (PAVE) Study: a pilot multicentered, randomized, controlled trial to study effects of periodontal therapy in a secondary prevention model of cardiovascular disease.
In the Periodontitis and Vascular Events (PAVE) pilot study, periodontal therapy was provided as an intervention in a secondary cardiac event prevention model through five coordinated cardiac-dental centers. Subjects were randomized to either community care or protocol provided scaling and root planing to evaluate effects on periodontal status and systemic levels of high-sensitivity C-reactive protein (hs-CRP). After 6 months, there was a significant reduction in mean probing depth and extent of 4- or 5-mm pockets. However, there were no significant differences in attachment levels, bleeding upon probing, or extent of subgingival calculus comparing subjects assigned to protocol therapy (n = 151) to those assigned to community care (n = 152). Using intent-to-treat analyses, there was no significant effect on serum hs-CRP levels at 6 months. However, 48% of the subjects randomized to community care received preventive or periodontal treatments. Secondary analyses demonstrated that consideration of any preventive or periodontal care (i.e., any treatment) compared to no treatment showed a significant reduction in the percentage of people with elevated hs-CRP (values >3 mg/l) at 6 months. However, obesity nullified the periodontal treatment effects on hs-CRP reduction. The adjusted odds ratio for hs-CRP levels >3 mg/l at 6 months for any treatment versus no treatment among non-obese individuals was 0.26 (95% confidence interval: 0.09 to 0.72), adjusting for smoking, marital status, and gender. This pilot study demonstrated the critical role of considering obesity as well as rigorous preventive and periodontal care in trials designed to reduce cardiovascular risk.


IPOP: An Integrative Plant Multi-omics Platform for Cross-species Comparison and Evolutionary Study.
The advent of high-throughput sequencing technologies has led to the production of a significant amount of omics data in plants, which serves as valuable assets for conducting cross-species multi-omics comparative analysis. Nevertheless, the current dearth of comprehensive platforms providing evolutionary annotation information and multi-species multi-omics data impedes users from systematically and efficiently performing evolutionary and functional analysis on specific genes. In order to establish an advanced plant multi-omics platform that provides timely, accurate, and high-caliber omics information, we collected 7 distinct types of omics data from 6 monocots, 6 dicots, and 1 moss, and reanalyzed these data using standardized pipelines. Additionally, we furnished homology information, duplication events, and phylostratigraphic stages of 13 species to facilitate evolutionary examination. Furthermore, the integrative plant omics platform (IPOP) is bundled with a variety of online analysis tools that aid users in conducting evolutionary and functional analysis. Specifically, the Multi-omics Integration Analysis tool is available to consolidate information from diverse omics sources, while the Transcriptome-wide Association Analysis tool facilitates the linkage of functional analysis with phenotype. To illustrate the application of IPOP, we conducted a case study on the YTH domain gene family, wherein we observed shared functionalities within orthologous groups and discerned variations in evolutionary patterns across these groups. To summarize, the IPOP platform offers valuable evolutionary insights and multi-omics data to the plant sciences community, effectively addressing the need for cross-species comparison and evolutionary research platforms. All data and modules within IPOP are freely accessible for academic purposes (http://omicstudio.cloud:4012/ipod/).


Transient stiffening of mitral valve leaflets in the beating heart.
Anterior mitral leaflet stiffness during isovolumic contraction (IVC) is much greater than that during isovolumic relaxation (IVR). We have hypothesized that this stiffening is due to transient early systolic force development in the slip of cardiac myocytes in the annular third of the anterior leaflet. Because the atrium is excited before IVC and leaflet myocytes contract for < or = 250 ms, this hypothesis predicts that IVC leaflet stiffness will drop to near-IVR values in the latter half of ventricular systole. We tested this prediction using radiopaque markers and inverse finite element analysis of 30 beats in 10 ovine hearts. For each beat, circumferential (E(c)) and radial (E(r)) stiffness was determined during IVC (Deltat(1)), end IVC to midsystole (Deltat(2)), midsystole to IVR onset (Deltat(3)), and IVR (Deltat(4)). Group mean stiffness (E(c) + or - SD; E(r) + or - SD; in N/mm(2)) during Deltat(1) (44 + or - 16; 15 + or - 4) was 1.6-1.7 times that during Deltat(4) (28 + or - 11; 9 + or - 3); Deltat(2) stiffness (39 + or - 15; 14 + or - 4) was 1.3-1.5 times that of Deltat(4), but Deltat(3) stiffness (32 + or - 12; 11 + or - 3) was only 1.1-1.2 times that of Deltat(4). The stiffness drop during Deltat(3) supports the hypothesis that anterior leaflet stiffening during IVC arises primarily from transient force development in leaflet cardiac myocytes, with stiffness reduced as this leaflet muscle relaxes in the latter half of ventricular systole.


Differential contribution of FXa and thrombin to vascular inflammation in a mouse model of sickle cell disease.
Activation of coagulation and vascular inflammation are prominent features of sickle cell disease (SCD). Previously, we have shown that inhibition of tissue factor (TF) attenuates activation of coagulation and vascular inflammation in mouse models of SCD. In this study, we examined the mechanism by which coagulation proteases enhance vascular inflammation in sickle BERK mice. To specifically investigate the contribution of FXa and thrombin, mice were fed chow containing either rivaroxaban or dabigatran, respectively. In addition, we used bone marrow transplantation to generate sickle mice deficient in either protease activated receptor-1 (PAR-1) or protease activated receptor-2 (PAR-2) on nonhematopoietic cells. FXa inhibition and PAR-2 deficiency in nonhematopoietic cells attenuated systemic inflammation, measured by plasma levels of interleukin-6 (IL-6). In contrast, neither thrombin inhibition nor PAR-1 deficiency in nonhematopoietic cells affected plasma levels of IL-6 in sickle mice. However, thrombin did contribute to neutrophil infiltration in the lung, independently of PAR-1 expressed by nonhematopoietic cells. Furthermore, the TF-dependent increase in plasma levels of soluble vascular cell adhesion molecule-1 in sickle mice was not mediated by FXa or thrombin. Our data indicate that TF, FXa, and thrombin differentially contribute to vascular inflammation in a mouse model of SCD.


Comprehensive analysis of PPARalpha-dependent regulation of hepatic lipid metabolism by expression profiling.
PPARalpha is a ligand-activated transcription factor involved in the regulation of nutrient metabolism and inflammation. Although much is already known about the function of PPARalpha in hepatic lipid metabolism, many PPARalpha-dependent pathways and genes have yet to be discovered. In order to obtain an overview of PPARalpha-regulated genes relevant to lipid metabolism, and to probe for novel candidate PPARalpha target genes, livers from several animal studies in which PPARalpha was activated and/or disabled were analyzed by Affymetrix GeneChips. Numerous novel PPARalpha-regulated genes relevant to lipid metabolism were identified. Out of this set of genes, eight genes were singled out for study of PPARalpha-dependent regulation in mouse liver and in mouse, rat, and human primary hepatocytes, including thioredoxin interacting protein (Txnip), electron-transferring-flavoprotein beta polypeptide (Etfb), electron-transferring-flavoprotein dehydrogenase (Etfdh), phosphatidylcholine transfer protein (Pctp), endothelial lipase (EL, Lipg), adipose triglyceride lipase (Pnpla2), hormone-sensitive lipase (HSL, Lipe), and monoglyceride lipase (Mgll). Using an in silico screening approach, one or more PPAR response elements (PPREs) were identified in each of these genes. Regulation of Pnpla2, Lipe, and Mgll, which are involved in triglyceride hydrolysis, was studied under conditions of elevated hepatic lipids. In wild-type mice fed a high fat diet, the decrease in hepatic lipids following treatment with the PPARalpha agonist Wy14643 was paralleled by significant up-regulation of Pnpla2, Lipe, and Mgll, suggesting that induction of triglyceride hydrolysis may contribute to the anti-steatotic role of PPARalpha. Our study illustrates the power of transcriptional profiling to uncover novel PPARalpha-regulated genes and pathways in liver.


Safety and recommendations for TMS use in healthy subjects and patient populations, with updates on training, ethical and regulatory issues: Expert Guidelines.
This article is based on a consensus conference, promoted and supported by the International Federation of Clinical Neurophysiology (IFCN), which took place in Siena (Italy) in October 2018. The meeting intended to update the ten-year-old safety guidelines for the application of transcranial magnetic stimulation (TMS) in research and clinical settings (Rossi et al., 2009). Therefore, only emerging and new issues are covered in detail, leaving still valid the 2009 recommendations regarding the description of conventional or patterned TMS protocols, the screening of subjects/patients, the need of neurophysiological monitoring for new protocols, the utilization of reference thresholds of stimulation, the managing of seizures and the list of minor side effects. New issues discussed in detail from the meeting up to April 2020 are safety issues of recently developed stimulation devices and pulse configurations; duties and responsibility of device makers; novel scenarios of TMS applications such as in the neuroimaging context or imaging-guided and robot-guided TMS; TMS interleaved with transcranial electrical stimulation; safety during paired associative stimulation interventions; and risks of using TMS to induce therapeutic seizures (magnetic seizure therapy). An update on the possible induction of seizures, theoretically the most serious risk of TMS, is provided. It has become apparent that such a risk is low, even in patients taking drugs acting on the central nervous system, at least with the use of traditional stimulation parameters and focal coils for which large data sets are available. Finally, new operational guidelines are provided for safety in planning future trials based on traditional and patterned TMS protocols, as well as a summary of the minimal training requirements for operators, and a note on ethics of neuroenhancement.


Sub-Nanomolar Sensitivity of Nitric Oxide Mediated Regulation of cGMP and Vasomotor Reactivity in Vascular Smooth Muscle.
Nitric oxide (NO) is a potent dilator of vascular smooth muscle (VSM) by modulating intracellular cGMP ([cGMP](i)) through the binding and activation of receptor guanylyl cylases (sGC). The kinetic relationship of NO and sGC, as well as the subsequent regulation of [cGMP](i) and its effects on blood vessel vasodilation, is largely unknown. In isolated VSM cells exposed to both pulsed and clamped NO we observed transient and sustained increases in [cGMP](i), with sub-nanomolar sensitivity to NO (EC(50) = 0.28 nM). Through the use of pharmacological inhibitors of sGC, PDE5, and PKG, a comprehensive VSM-specific modeling algorithm was constructed to elucidate the concerted activity profiles of sGC, PDE5, phosphorylated PDE5, and PDE1 in the maintenance of [cGMP](i). In small pressure-constricted arteries of the resistance vasculature we again observed both transient and sustained relaxations upon delivery of pulsed and clamped NO, while maintaining a similarly high sensitivity to NO (EC(50) = 0.42 nM). Our results propose an intricate dependency of the messengers and enzymes involved in cGMP homeostasis, and vasodilation in VSM. Particularly, the high sensitivity of sGC to NO in primary tissue indicates how small changes in the concentrations of NO, irrespective of the form of NO delivery, can have significant effects on the dynamic regulation of vascular tone.


Monitoring risk factors of cardiovascular disease in cancer survivors.
There exist published literature for cardiovascular disease (CVD) risk monitoring in cancer survivors but the extent of monitoring in clinical oncology practice is unknown. We performed an interactive survey at a Royal College of Physicians conference (11 November 2016) attended by practitioners with an interest in late effects of cancer treatment and supplemented the survey with an audit among 32 lung cancer survivors treated at St Peter's NHS Hospital in 2012-2016. Among the practitioners, 40% reported CVD risk monitoring performed at least annually, which is compatible with European Group for Blood and Marrow Transplantation Guidelines, but 31% indicated that monitoring was never performed. In contrast, 77% felt that at least an annual assessment was required (p<0.001). Corroborating these data, among the lung cancer survivors, 31% and 16% had lipids or glucose/HbA1C measured annually, and 28% and 31% had never had these tests performed since their cancer treatment. Alerting healthcare providers to review protocols may help reduce CVD after cancer treatments.


Can occlusal evaluation of children with unilateral cleft lip and palate help determine future maxillofacial morphology?
Objective : To investigate the change in occlusal evaluations from the 5-year-olds' index to the Goslon Yardstick and to compare the relationship between the evaluations and maxillofacial growth in patients with complete unilateral cleft lip and palate (UCLP). Design : A prospective longitudinal study. Subjects : The sample consisted of 85 patients with complete UCLP who underwent surgery from 1969 to 1994 and were treated at the Kyushu University Hospital in Fukuoka, Japan. Subjects had two serial dental casts performed at the ages of 5 and 10 years. Furthermore, each patient had lateral cephalographs taken at the age of 5 years, 76 of 85 subjects had films taken at the age of 10 years, and 54 subjects also had lateral cephalograms taken after the age of 15 years. Methods : Every dental cast was evaluated by the 5-year-olds' index and the Goslon Yardstick, respectively. The lateral cephalographs were traced and digitized, and angular dimensions were calculated. Outcomes were compared using Spearman's rank-order correlation analysis and the Kruskal-Wallis analysis. Results and Conclusion : Dental arch relationships were evaluated and rated as 2.96 in the 5-year-olds' index and 2.85 in the Goslon Yardstick, respectively. Both groupings showed a significant relationship, and they showed no change in 36 out of 85 subjects (42.3%), significant improvement in 30 (35.3%), and deterioration in 19 (22.3%). Two occlusal groupings and maxillofacial morphology on the cephalographs indicated that the grouping reflected the anteroposterior position of the mandible. Moreover, both groupings showed some relation to previous maxillofacial growth, but they did not show any relationship with future growth. The Goslon Yardstick may not predict maxillofacial morphology in adulthood.


Biotin responsive multiple carboxylase deficiency presenting as diabetic ketoacidosis.
Multiple carboxylase deficiency (MCD) is a rare inherited metabolic disease of biotin dependency due to deficiency of holocarboxylase synthetase (HCS) or biotinidase deficiency. A 30-month-old female patient who presented with the initial features of diabetic ketoacidosis (severe metabolic acidosis, ketosis, and hyperglycemia), lactic acidemia, moderate hyperammonemia, and generalized organic aciduria is described. Associated symptoms and signs included erythematous skin rashes, alopecia and developmental delay. The patient responded dramatically to treatment with biotin (10 mg/day) showing normalization of clinical symptoms and most biochemical abnormalities. Based on the urine organic profile by gas chromatography/ mass spectrometry (GC/MS), the diagnosis of MCD was made. A plasma tandem mass study confirmed this diagnosis. The biotinase activity in serum was normal, indicating that this was a rare case of late-onset HCS deficiency.


Do drug users use less alcohol than non-drug users? A comparison of ethyl glucuronide concentrations in hair between the two groups in medico-legal cases.
Two groups were selected from the remainder of hair samples that had been tested for drugs at TrichoTech for medico-legal cases: samples that tested negative (drug-negative group; N=42, age 33.4+/-7.2 years) and samples that tested positive for drugs (drug-positive group; N=57, age 32.5+/-8.8 years). A rapid, simple method to detect the ethanol metabolite, ethyl glucuronide (EtG) in hair has been developed. The hair samples were sectioned, and then submitted to overnight sonication in water. Samples then underwent SPE using anion exchange cartridges, followed by derivatisation with N,O-bis[trimethylsilyl]trifluoroacetamide (BSTFA), before confirmation by GC-MS/MS. The assay produced excellent linearity and sensitivity over the calibration range 0.02-1.0 ng/mg, assuming a 10 mg hair sample. The mean age of the two groups was not statistically different (p=0.575, Student t-test), indicating a homogeneous group. Twelve of the 57 (21.0%) hair samples of the drug-positive group tested positive for EtG, and 17 of the 42 (40.5%) hair samples of the drug-negative group tested positive for EtG. The mean concentration of EtG in the drug-positive group was 0.011 ng/mg compared to 0.107 ng/mg in the drug-negative group. When the full results of this study were subjected to statistical analysis it was shown that EtG levels in the drug-negative group were statistically higher than those found in the drug-positive group (p<0.05). This preliminary finding may be of use in the study of addiction and adds valuable data to previous studies regarding the use of EtG as a valuable marker for alcohol levels in hair.


[Ultrastructure of a granular-cell breast tumor].
Electron microscopic examination of a rare granular cell tumour of the mammary gland that provoked difficulties in the diagnosis was performed. Ultrastructural organization of the tumour allowed one to establish a proper diagnosis and to confirm the hypothesis on the origin of this tumour from Schwann's cells.


[Psychiatry : Swiss recommendations for treatment].
The treatment of psychiatric disorders has been subject of numerous research, which are at the basis of international guidelines and recommendations. The Swiss Society for Psychiatry and Psychotherapy has decided to mandate various expert groups to develop Swiss recommendations. Such recommendations are now available for the main psychiatric disorders and this paper summarizes their main features in order to make them more visible.


Neoantigen Dendritic Cell Vaccination Combined with Anti-CD38 and CpG Elicits Anti-Tumor Immunity against the Immune Checkpoint Therapy-Resistant Murine Lung Cancer Cell Line LLC1.
An important factor associated with primary resistance to immune-checkpoint therapies (ICT) is a "cold" tumor microenvironment (TME), characterized by the absence of T cell infiltration and a non-inflammatory milieu. Whole-exome and RNA sequencing to predict neoantigen expression was performed on the LLC1 cell line which forms "cold" tumors in mice. Dendritic cell (DC)-based vaccination strategies were developed using candidate neoantigen long peptides (LPs). A total of 2536 missense mutations were identified in LLC1 and of 132 candidate neoantigen short peptides, 25 were found to induce CD8+ T cell responses. However, they failed to inhibit LLC1 growth when incorporated into a cancer vaccine. In contrast, DCs pulsed with LPs induced CD4+ and CD8+ T cell responses and one of them, designated L82, delayed LLC1 growth in vivo. By RNA-Seq, CD38 was highly expressed by LLC1 tumor cells and, therefore, anti-CD38 antibody treatment was combined with L82-pulsed DC vaccination. This combination effectively suppressed tumor growth via a mechanism relying on decreased regulatory T cells in the tumor. This study demonstrated that an appropriate vaccination strategy combining neoantigen peptide-pulsed DC with anti-CD38 antibody can render an ICT-resistant "cold" tumor susceptible to immune rejection via a mechanism involving neutralization of regulatory T cells.


Elevated plasma homocysteine leads to alterations in fibrin clot structure and stability: implications for the mechanism of thrombosis in hyperhomocysteinemia.
Elevated plasma homocysteine is associated with an increased risk of atherosclerosis and thrombosis. However, the mechanisms by which homocysteine might cause these events are not understood. We hypothesized that hyperhomocysteinemia might lead to modification of fibrinogen in vivo, thereby causing altered fibrin clot structure. New Zealand White rabbits were injected intraperitoneally (i.p.) every 12 h through an indwelling catheter with homocysteine or buffer for 8 weeks. This treatment raised the plasma homocysteine levels to about 30 micro mol L(-1) compared with 13.5 micro mol L(-1) in control rabbits by the end of the treatment period. The fibrinogen levels were 3.2 +/- 0.6 in homocysteine-treated and 2.5 +/- 1.1 mg mL(-1) in control rabbits. The reptilase time was prolonged to 363 +/- 88 for plasma from homocysteine-treated rabbits compared with 194 +/- 48 s for controls (P < 0.01). The thrombin clotting time (TCT) for the homocysteine-treated rabbits was significantly shorter, 7.5 +/- 1.7 compared with 28.6 +/- 18 s for the controls (P < 0.05). The calcium dependence of the thrombin clotting time was also different in homocysteinemic and control plasmas. Clots from plasma or fibrinogen of homocysteinemic rabbits were composed of thinner fibers than control clots. The clots formed from purified fibrinogen from homocysteine-treated rabbits were lyzed more slowly by plasmin than comparable clots from control fibrinogen. Congenital dysfibrinogenemias have been described that are associated with fibrin clots composed of thin, tightly packed fibers that are abnormally resistant to fibrinolysis, and recurrent thrombosis. Our results suggest that elevated plasma homocysteine leads to a similar acquired dysfibrinogenemia. The formation of clots that are abnormally resistant to fibrinolysis could directly contribute to the increased risk of thrombosis in hyperhomocysteinemia.


Photoexpulsion of surface-grafted ruthenium complexes and subsequent release of cytotoxic cargos to cancer cells from mesoporous silica nanoparticles.
Ruthenium(II) polypyridyl complexes have emerged both as promising probes of DNA structure and as anticancer agents because of their unique photophysical and cytotoxic properties. A key consideration in the administration of those therapeutic agents is the optimization of their chemical reactivities to allow facile attack on the target sites, yet avoid unwanted side effects. Here, we present a drug delivery platform technology, obtained by grafting the surface of mesoporous silica nanoparticles (MSNPs) with ruthenium(II) dipyridophenazine (dppz) complexes. This hybrid nanomaterial displays enhanced luminescent properties relative to that of the ruthenium(II) dppz complex in a homogeneous phase. Since the coordination between the ruthenium(II) complex and a monodentate ligand linked covalently to the nanoparticles can be cleaved under irradiation with visible light, the ruthenium complex can be released from the surface of the nanoparticles by selective substitution of this ligand with a water molecule. Indeed, the modified MSNPs undergo rapid cellular uptake, and after activation with light, the release of an aqua ruthenium(II) complex is observed. We have delivered, in combination, the ruthenium(II) complex and paclitaxel, loaded in the mesoporous structure, to breast cancer cells. This hybrid material represents a promising candidate as one of the so-called theranostic agents that possess both diagnostic and therapeutic functions.


Pre- and post-natal growth and protein turnover in the lung of the rat.
The growth of the rat lung was studied at six ages, from 18 days of fetal life to old age (i.e. 105 weeks). Most of the increase in lung size appeared to involve cellular hyperplasia rather than hypertrophy, the DNA content of the lung increasing 96-fold from one extreme of life to the other. Pulmonary rates of protein turnover were high and were, age for age, consistently greater than the rates in the whole body. The age-related decline in the rate of lung growth corresponded to a marked decrease in the fractional rate of protein synthesis, i.e. from 93 to 33% per day during fetal and neonatal life. This in turn correlated with a 58% fall in the ribosomal capacity. From weaning onwards, synthesis rates remained between 30 and 40% per day. In contrast, the degradation of lung proteins was unchanged, at 28-38% per day throughout both fetal and post-natal life.


Modulation of outer hair cell compliance and force by agents that affect hearing.
Force generated by outer hair cells is thought to be an essential source of mechanical input to the normal cochlea. Many disease processes in the inner ear act via outer hair cells. It is therefore plausible that such disease processes modulate outer hair cell force generation. The force generated by an isolated, electrically stimulated outer hair cell against a load may be represented by an intrinsic motor and a passive axial stiffness in series. Thus modulation of outer hair cell force generation may occur either by action on the motor or indirectly by an action on cell stiffness. In this study, the effects of agents that affect hearing on outer hair cell stiffness and force generation have been examined. Overstimulation and hypoosmotic challenge caused cells to decrease in length and increase in stiffness. The force generated by a constant voltage stimulus increased consequent to the stiffness increase. Hyperosmotic challenge elicited a stiffness decrease and a force decrease. In contrast, salicylate caused a decrease in force without stiffness change. The results suggest that outer hair cell force generation in vivo may be modulated in at least two ways.


Use of prostate systematic and targeted biopsy on the basis of multiparametric MRI in biopsy-naive patients (MRI-FIRST): a prospective, multicentre, paired diagnostic study.
Whether multiparametric MRI improves the detection of clinically significant prostate cancer and avoids the need for systematic biopsy in biopsy-naive patients remains controversial. We aimed to investigate whether using this approach before biopsy would improve detection of clinically significant prostate cancer in biopsy-naive patients. In this prospective, multicentre, paired diagnostic study, done at 16 centres in France, we enrolled patients aged 18-75 years with prostate-specific antigen concentrations of 20 ng/mL or less, and with stage T2c or lower prostate cancer. Eligible patients had been referred for prostate multiparametric MRI before a first set of prostate biopsies, with a planned interval of less than 3 months between MRI and biopsies. An operator masked to multiparametric MRI results did a systematic biopsy by obtaining 12 systematic cores and up to two cores targeting hypoechoic lesions. In the same patient, another operator targeted up to two lesions seen on MRI with a Likert score of 3 or higher (three cores per lesion) using targeted biopsy based on multiparametric MRI findings. Patients with negative multiparametric MRI (Likert score ≤2) had systematic biopsy only. The primary outcome was the detection of clinically significant prostate cancer of International Society of Urological Pathology grade group 2 or higher (csPCa-A), analysed in all patients who received both systematic and targeted biopsies and whose results from both were available for pathological central review, including patients who had protocol deviations. This study is registered with ClinicalTrials.gov, number NCT02485379, and is closed to new participants. Between July 15, 2015, and Aug 11, 2016, we enrolled 275 patients. 24 (9%) were excluded from the analysis. 53 (21%) of 251 analysed patients had negative (Likert ≤2) multiparametric MRI. csPCa-A was detected in 94 (37%) of 251 patients. 13 (14%) of these 94 patients were diagnosed by systematic biopsy only, 19 (20%) by targeted biopsy only, and 62 (66%) by both techniques. Detection of csPCa-A by systematic biopsy (29·9%, 95% CI 24·3-36·0) and targeted biopsy (32·3%, 26·5-38·4) did not differ significantly (p=0·38). csPCa-A would have been missed in 5·2% (95% CI 2·8-8·7) of patients had systematic biopsy not been done, and in 7·6% (4·6-11·6) of patients had targeted biopsy not been done. Four grade 3 post-biopsy adverse events were reported (3 cases of prostatitis, and 1 case of urinary retention with haematuria). There was no difference between systematic biopsy and targeted biopsy in the detection of ISUP grade group 2 or higher prostate cancer; however, this detection was improved by combining both techniques and both techniques showed substantial added value. Thus, obtaining a multiparametric MRI before biopsy in biopsy-naive patients can improve the detection of clinically significant prostate cancer but does not seem to avoid the need for systematic biopsy. French National Cancer Institute.


Adaptive radiosurgery based on two simultaneous dose prescriptions in the management of large renal cell carcinoma brain metastases in critical areas: Towards customization.
The long-term benefits of local therapy in metastatic renal cell carcinoma (mRCC) have been widely documented. In this context, single fraction gamma knife radiosurgery (SF-GKRS) is routinely used in the management of brain metastases. However, SF-GKRS is not always feasible due to volumetric and regional constraints. We intend to illustrate how a dose-volume adaptive hypofractionated GKRS technique based on two concurrent dose prescriptions termed rapid rescue radiosurgery (RRR) can be utilized in this particular scenario. A 56-year-old man presented with left-sided hemiparesis; the imaging showed a 13.1 cc brain metastasis in the right central sulcus (Met 1). Further investigation confirmed the histology to be a metastatic clear cell RCC. Met 1 was treated with upfront RRR. Follow-up magnetic resonance imaging (MRI) at 10 months showed further volume regression of Met 1; however, concurrently, a new 17.3 cc lesion was reported in the boundaries of the left frontotemporal region (Met 2) as well as a small metastasis (<1 cc) in the left temporal lobe (Met 3). Met 2 and Met 3 underwent RRR and SF-GKRS, respectively. Gradual and sustained tumor ablation of Met 1 and Met 2 was demonstrated on a 20 months long follow- up. The patient succumbed to extracranial disease 21 months after the treatment of Met 1 without evidence of neurological impairment post-RRR. Despite poor prognosis and precluding clinical factors (failing systemic treatment, eloquent location, and radioresistant histology), RRR provided optimal tumor ablation and salvage of neurofunction with limited toxicity throughout follow-up.


A first-principles study on the phase stability and physical properties of a B-site ordered Nd2CrFeO6 double perovskite.
Here, the first-principles predictions on the structural stability, magnetic behavior and electronic structure of B-site ordered double perovskite Nd2CrFeO6 have been reported. Initially, the ground state of the parent single perovskites NdCrO3 and NdFeO3 has been studied to determine the relevant Hubbard U parameter to investigate the properties of Nd2CrFeO6. The thermodynamic, mechanical, and dynamic stability analyses suggest the possibility of the synthesis of the Nd2CrFeO6 double perovskite at ambient pressure. The compound shows a ferrimagnetic nature with 2 μB net magnetic moment and the magnetic ordering temperature has been estimated to be ∼265 K. The electronic structure indicates a higher probability of direct photon transition over the indirect transition with a band gap of ∼1.85 eV. Additional effects of Nd (4f) spin and spin-orbit coupling on the band edges have been found to be negligible for this 4f-3d-3d spin system. This first-principles investigation predicts that due to the ferrimagnetic nature and a significantly lower band gap compared to those of its antiferromagnetic parent single perovskites, the B-site ordered Nd2CrFeO6 double perovskite could be a promising material for spintronic and visible-light driven energy applications.


The Nutritional Composition and Energy Content of Donor Human Milk: A Systematic Review.
The American Academy of Pediatrics recommends donor human milk (DHM) as the preferred feeding strategy for preterm infants when the milk of the mother is unavailable, based on conclusive evidence of lower rates of necrotizing enterocolitis with DHM feedings compared with preterm infant formula. The nutritional composition of DHM may differ from maternal milk for many reasons including differences in maternal characteristics, milk collection methods, and the impact of donor milk banking practices. The purpose of this systematic review is to examine the literature regarding research on the fat, protein, carbohydrate, vitamin, and mineral composition of DHM obtained through nonprofit milk banks or commercial entities. PubMed, CINAHL, and Scopus databases were searched for articles published between 1985 and 30 April, 2019. In total, 164 abstracts were screened independently by 2 investigators, and 14 studies met all inclusion criteria. Studies were predominantly small (<50 samples) and measured macronutrients. Few studies assessed vitamins and minerals. Information bias was prevalent due to the use of a variety of analytical methods which influence accuracy and cross-study comparisons. Other sources of information bias included missing information regarding methods for protein and calorie assessment. Despite these limitations, existing research suggests the potential for 2-fold and greater differences in the fat, protein, and energy composition of DHM, with mean values for energy and fat often below clinical reference values expected for human milk. Further research is warranted regarding the nutritional composition of DHM, with a prioritization on measuring macronutrients and micronutrients using established reference methods.


Molecular Epidemiology, Natural History, and Long-Term Outcomes of Multidrug-Resistant Enterobacterales Colonization and Infections Among Solid Organ Transplant Recipients.
Multidrug-resistant Enterobacterales (MDR-E), including carbapenem-resistant and third-generation cephalosporin-resistant Enterobacterales (CRE, CefR-E), are major pathogens following solid organ transplantation (SOT). We prospectively studied patients who underwent lung, liver, and small bowel transplant from February 2015 through March 2017. Weekly perirectal swabs (up to 100 days post-transplant) were cultured for MDR-E. Whole-genome sequencing (WGS) was performed on gastrointestinal (GI) tract-colonizing and disease-causing isolates. Twenty-five percent (40 of 162) of patients were MDR-E GI-colonized. Klebsiella pneumoniae was the most common CRE and CefR-E. Klebsiella pneumoniae carbapenemases and CTX-M were leading causes of CR and CefR, respectively. Thirty-five percent of GI colonizers developed MDR-E infection vs 2% of noncolonizers (P < .0001). The attack rate was higher among CRE colonizers than CefR-E colonizers (53% vs 21%, P = .049). GI colonization and high body mass index were independent risk factors for MDR-E infection (P ≤ .004). Thirty-day mortality among infected patients was 6%. However, 44% of survivors developed recurrent infections; 43% of recurrences were late (285 days to 3.9 years after the initial infection). Long-term survival (median, 4.3 years post-transplant) did not differ significantly between MDR-E-infected and MDR-E-noninfected patients (71% vs 77%, P = .56). WGS phylogenetic analyses revealed that infections were caused by GI-colonizing strains and suggested unrecognized transmission of novel clonal group-258 sublineage CR-K. pneumoniae and horizontal transfer of resistance genes. MDR-E GI colonization was common following SOT and predisposed patients to infections by colonizing strains. MDR-E infections were associated with low short- and long-term mortality, but recurrences were frequent and often occurred years after initial infections. Findings provide support for MDR-E surveillance in our SOT program.


FvSNF1, the sucrose non-fermenting protein kinase gene of Fusarium virguliforme, is required for cell-wall-degrading enzymes expression and sudden death syndrome development in soybean.
Fusarium virguliforme is a soil-borne pathogenic fungus that causes sudden death syndrome (SDS) in soybean. Its pathogenicity is believed to require the activity of cell-wall-degrading enzymes (CWDEs). The sucrose non-fermenting protein kinase 1 gene (SNF1) is a key component of the glucose de-repression pathway in yeast, and a regulator of gene expression for CWDEs in some plant pathogenic fungi. To elucidate the functional role of the SNF1 homolog in F. virguliforme, FvSNF1 was disrupted using a split-marker strategy. Disruption of FvSNF1 in F. virguliforme abolishes galactose utilization and causes poor growth on xylose, arabinose and sucrose. However, the resulting Fvsnf1 mutant grew similar to wild-type and ectopic transformants on glucose, fructose, maltose, or pectin as the main source of carbon. The Fvsnf1 mutant displayed no expression of the gene-encoding galactose oxidase (GAO), a secretory enzyme that catalyzes oxidation of D-galactose. It also exhibited a significant reduction in the expression of several CWDE-coding genes in contrast to the wild-type strain. Greenhouse pathogenicity assays revealed that the Fvsnf1 mutant was severely impaired in its ability to cause SDS on challenged soybean plants. Microscopy and microtome studies on infected roots showed that the Fvsnf1 mutant was defective in colonizing vascular tissue of infected plants. Cross and longitudinal sections of infected roots stained with fluorescein-labeled wheat germ agglutinin and Congo red showed that the Fvsnf1 mutant failed to colonize the xylem vessels and phloem tissue at later stages of infection. Quantification of the fungal biomass in inoculated roots further confirmed a reduced colonization of roots by the Fvsnf1 mutant when compared to the wild type. These findings suggest that FvSNF1 regulates the expression of CWDEs in F. virguliforme, thus affecting the virulence of the fungus on soybean.


Use of administrative medical databases in population-based research.
Administrative medical databases are massive repositories of data collected in healthcare for various purposes. Such databases are maintained in hospitals, health maintenance organisations and health insurance organisations. Administrative databases may contain medical claims for reimbursement, records of health services, medical procedures, prescriptions, and diagnoses information. It is clear that such systems may provide a valuable variety of clinical and demographic information as well as an on-going process of data collection. In general, information gathering in these databases does not initially presume and is not planned for research purposes. Nonetheless, administrative databases may be used as a robust research tool. In this article, we address the subject of public health research that employs administrative data. We discuss the biases and the limitations of such research, as well as other important epidemiological and biostatistical key points specific to administrative database studies.


EIF2α/ATF4 pathway enhances proliferation of mesangial cell via cyclin D1 during endoplasmic reticulum stress in IgA nephropathy.
IgA nephropathy (IgAN) is an essential cause of kidney failure and end-stage kidney disease worldwide. Mesangial hypercellularity is an important characteristic of IgAN, but the underlying mechanism remains unclear. Endoplasmic reticulum (ER) stress is a series of stress responses to restore the function of endoplasmic reticulum. We aimed to explore how ER stress functioned in kidneys of IgAN. We first examined ER stress in IgAN kidneys in vivo and in vitro, by testing the levels of ER stress associated proteins (BIP, p-eIF2α and ATF4). Our results showed that ER stress was activated in IgAN patients, mice and cell model. ER stress activation was related to the distribution of IgA deposition and the degree of mesangial proliferation. To determine the role of ER stress in mesangial cell (MC) proliferation of IgAN, we then tested the levels of ER stress and MC proliferation (cyclin D1, cell viability and cell cycle) through inhibiting ER stress associated proteins. After inhibiting ER stress associated proteins, ER stress was inactivated and cell proliferation was inhibited in MCs. We also explored the correlation between ER stress in the glomerulus and the clinical outcomes of IgAN patients in a prospective study. Patients with lower expression of p-eIF2α or ATF4 had higher rates of hematuria remission, proteinuria remission and clinical remission. In summary, our work outlines that in IgAN, ER stress mediated by eIF2α/ATF4 pathway promotes MC proliferation via up-regulating the expression of cyclin D1. Furthermore, p-eIF2α and ATF4 in the glomerulus negatively correlate with the clinical remission of IgAN patients.


New cephalosporins with 7-acyl groups derived from beta-ketoacids. I. 7-(beta-Ketoacylamino)cephalosporins.
The synthesis and antimicrobial profile of a series of 7-(beta-ketoacylamino)cephalosporins (1) bearing an acetoxymethyl or a heterocyclichiomethyl group at the 3-position are described. Of this series, 3-[[(1-methyl-1H-tetrazol-5-yl)thio]methyl]-7-(3-oxobutyrylamino)ceph-3-em-4-carboxylic acid (11) showed moderate antibacterial activities in in vitro and in vivo tests.


Why young adults use tobacco-free nicotine E-cigarettes: An analysis of qualitative data.
Tobacco-free nicotine (TFN) is often marketed as being better tasting and healthier than tobacco-derived nicotine (TDN). Studies have yet to systematically characterize subjective experiences using TFN e-cigarettes. This qualitative study explored young adults' reasons for using TFN e-cigarettes and perceptions of differences between using TFN and TDN e-cigarettes. We conducted a national online Qualtrics survey of U.S. young adults aged 18-25 years in October 2021. Respondents who reported lifetime TFN e-cigarette use (n = 317, mean age: 20.8 (SD:2.2), 53.9 % female) provided write-in responses to open-ended questions assessing reasons for using TFN e-cigarettes and perceptions of differences between TFN and TDN e-cigarettes. Responses from participants were thematically analyzed by independent raters. Reasons for using TFN e-cigarettes included product taste/flavor, health reasons (e.g. believing TFN was healthy), and product accessibility. Perceptions of differences between TFN and TDN e-cigarettes included differences in taste/ flavor and acute psychoactive and physical effects (e.g., nicotine buzz, throat hit), with TFN e-cigarettes being described as generally stronger and superior relative to TDN e-cigarettes. These qualitative data suggest that taste and perceived health benefits are primary reasons that young adults try TFN e-cigarettes. Compared to TDN, young adults perceive TFN as better tasting with different psychoactive and less aversive effects. Determining if these perceived reasons for use and differences occur in blinded human experimental studies is a necessary next step to understanding TFN e-cigarette use. Additionally, understanding why young adults use TFN e-cigarettes is important in the development of prevention communication about these products.


Near-infrared transmittance pulse oximetry with laser diodes.
Pulse oximeters are widely used for noninvasive monitoring of oxygen saturation in arterial blood hemoglobin. We present a transmittance pulse oximetry system based on near-infrared (NIR) laser diodes (750 and 850 nm) for monitoring oxygen saturation of arterial blood hemoglobin. The pulse oximetry system is made up of the optical sensor, sensor electronics, and processing block. Also, we show experimental results obtained during the development of the whole NIR transmittance pulse oximetry system along with modifications in the sensor configuration, signal processing algorithm, and calibration procedure. Issues concerning wavelength selection and its implications for the improvement of the transmittance pulse oximetry technique are discussed. The results obtained demonstrate the proposed system's usefulness in monitoring a wide range of oxygen saturation levels.


Arm-less mitochondrial tRNAs conserved for over 30 millions of years in spiders.
In recent years, Next Generation Sequencing (NGS) has accelerated the generation of full mitogenomes, providing abundant material for studying different aspects of molecular evolution. Some mitogenomes have been observed to harbor atypical sequences with bizarre secondary structures, which origins and significance could only be fully understood in an evolutionary framework. Here we report and analyze the mitochondrial sequences and gene arrangements of six closely related spiders in the sister genera Parachtes and Harpactocrates, which belong to the nocturnal, ground dwelling family Dysderidae. Species of both genera have compacted mitogenomes with many overlapping genes and strikingly reduced tRNAs that are among the shortest described within metazoans. Thanks to the conservation of the gene order and the nucleotide identity across close relatives, we were able to predict the secondary structures even on arm-less tRNAs, which would be otherwise unattainable for a single species. They exhibit aberrant secondary structures with the lack of either DHU or TΨC arms and many miss-pairings in the acceptor arm but this degeneracy trend goes even further since at least four tRNAs are arm-less in the six spider species studied. The conservation of at least four arm-less tRNA genes in two sister spider genera for about 30 myr suggest that these genes are still encoding fully functional tRNAs though they may be post-transcriptionally edited to be fully functional as previously described in other species. We suggest that the presence of overlapping and truncated tRNA genes may be related and explains why spider mitogenomes are smaller than those of other invertebrates.


Lowering gastrointestinal leak rates: a comparative analysis of robotic and laparoscopic gastric bypass.
Robotic-assisted surgery has been described for many general surgery procedures, including gastric bypass. This is a comparative study looking at the short-term outcomes and technical differences between laparoscopic Roux-en-Y gastric bypass (LRNY) and robotic-assisted Roux-en-Y gastric bypass (RARNY). Our database was reviewed for all Roux-en-Y gastric bypass procedures performed over the last 5 years. Operative times, length of stay, and all complications listed for the 90 days postoperatively were recorded and statistically analyzed. A total of 356 LRNY and 249 RARNY were performed. The average body mass index (BMI), age, and sex were similar between groups. On average, the RARNY took 17 min longer than the LRNY, this difference being significant (p < 0.01). Average length of stay for the two groups was similar (~3 days). There were a total of 51 complications in the standard laparoscopic group (14%), of which 14 (3.9%) were major complications. In the robotic group, there were 35 (14%) complications, of which 9 (3.6%) were major complications. The only significant difference in complication rate was for anastomotic leak at the gastrojejunostomy: there were no leaks in the robotic series, and six (1.7%) in the standard laparoscopic series (p = 0.04). Length of stay and overall complication rates were similar for RARNY and LRNY. There was no mortality in either group, and the complication rate was lower than literature standards. While the RARNY took longer, there was a significantly lower gastrointestinal leak rate in this group.


Physiologic responses and plasma endothelin-1 concentrations associated with abrupt cessation of nitric oxide inhalation in isoflurane-anesthetized horses.
To assess physiologic responses and plasma endothelin (ET)-1 concentrations associated with abrupt cessation of nitric oxide (NO) inhalation in isoflurane-anesthetized horses. 6 healthy adult Standardbreds. Horses were anesthetized with isoflurane in oxygen and placed in dorsal recumbency. Nitric oxide was pulsed into the respiratory tract for 2.5 hours, and then administration was abruptly discontinued. Just prior to commencement and at cessation of NO administration, and at intervals during a 30-minute period following cessation of NO inhalation, several variables including PaO(2), mean pulmonary artery pressure, venous admixture or pulmonary shunt fraction (Qs/Qt), and plasma ET-1 concentration were recorded or calculated. After cessation of NO inhalation, PaO(2) decreased slowly but significantly (172.7 +/- 29.8 mm Hg to 84.6 +/- 10.9 mm Hg) and Qs/Qt increased slowly but significantly (25 +/- 2% to 40 +/- 3%) over a 30-minute period. Mean pulmonary artery pressure increased slightly (14.0 +/- 1.3 mm Hg to 16.8 +/- 1 mm Hg) over the same time period. No change in serum ET-1 concentration was detected, and other variables did not change or underwent minor changes. The improvement in arterial oxygenation during pulsed inhalation of NO to healthy isoflurane-anesthetized horses decreased only gradually during a 30-minute period following cessation of NO inhalation, and serum ET-1 concentration was not affected. Because a rapid rebound response did not develop, inhalation of NO might be clinically useful in the treatment of hypoxemia in healthy isoflurane-anesthetized horses.


Multiomic disease signatures converge to cytotoxic CD8 T cells in primary Sjögren's syndrome.
Multiomics study was conducted to elucidate the crucial molecular mechanisms of primary Sjögren's syndrome (SS) pathology. We generated multiple data set from well-defined patients with SS, which includes whole-blood transcriptomes, serum proteomes and peripheral immunophenotyping. Based on our newly generated data, we performed an extensive bioinformatic investigation. Our integrative analysis identified SS gene signatures (SGS) dysregulated in widespread omics layers, including epigenomes, mRNAs and proteins. SGS predominantly involved the interferon signature and ADAMs substrates. Besides, SGS was significantly overlapped with SS-causing genes indicated by a genome-wide association study and expression trait loci analyses. Combining the molecular signatures with immunophenotypic profiles revealed that cytotoxic CD8 -T cells- were associated with SGS. Further, we observed the activation of SGS in cytotoxic CD8 T cells isolated from patients with SS. Our multiomics investigation identified gene signatures deeply associated with SS pathology and showed the involvement of cytotoxic CD8 T cells. These integrative relations across multiple layers will facilitate our understanding of SS at the system level.


Assessing cerebrovascular autoregulation from critical closing pressure and resistance area product during upright posture in aging and hypertension.
Static cerebral autoregulation (sCA) is believed to be resistant to aging and hypertensive pathology. However, methods to characterize autoregulation commonly rely on beat-by-beat mean hemodynamic measures and do not consider within-beat pulse wave characteristics that are impacted by arterial stiffening. We examined the role of critical closing pressure (CrCP) and resistance area product (RAP), two measures derived from the pulse wave, across supine lying, sitting, and standing postures in young adults, normotensive older adults, and older adults with controlled and uncontrolled hypertension (N = 80). Traditional measures of sCA, using both intracranial and extracranial methods, indicated similar efficiency across all groups, but within-beat measures suggested different mechanisms of regulation. At rest, RAP was increased in hypertension compared with young adults (P < 0.001), but CrCP was similar. In contrast, the drop in CrCP was the primary regulator of change in cerebrovascular resistance upon adopting an upright posture. Both CrCP and RAP demonstrated group-by-posture interaction effects (P < 0.05), with older hypertensive adults exhibiting a rise in RAP that was absent in other groups. The posture-related swings in CrCP and RAP were related to changes in both the pulsatile and mean components of arterial pressure, independent of age, cardiac output, and carbon dioxide. Group-by-posture differences in pulse pressure were mediated in part by an attenuated heart rate response in older hypertensive adults (P = 0.002). Examination of pulsatile measures in young, elderly, and hypertensive adults identified unique differences in how cerebral blood flow is regulated in upright posture.


The validity and reliability of the self-directed learning instrument (SDLI) in mainland Chinese nursing students.
Self-directed learning is crucial to the professional development of nursing students, and which enables them to expand the knowledge and enhance the quality of their practice. A validated self-directed learning instrument is important not only in assessing the individual's self-directed learning level, but also in evaluating the effectiveness of teaching or learning methods. The aim of this study is to evaluate the validity and reliability of the SDLI in mainland Chinese nursing students. A cross-sectional design with convenience sampling was used to recruit participants from three nursing schools. The mainland Chinese version of SDLI was tested with respect to validity and reliability in 1,499 nursing students, and another 30 nursing students were invited to evaluate the test-retest reliability of the scale in 7 days interval. Explorary factor analysis identified a four-factor structure, accounting for 56.101% of the total variance. The confirmatory factor analysis showed a good overall fit of this four-factor model. Convergent validity was supported by the highly positive Pearson's correlation between SDLI score and SRSSDL score (r = .876, p = .000). Cronbach's alpha for internal consistency of overall scale was .916, and 4 dimensions were between .755-.825.The test-retest reliability of overall scale was .850, and 4 dimensions were between .708-.821. The intraclass correlation coefficient (ICC) of overall scale was .916, and 4 dimensions were .822-.889. This study indicates that the SDLI is a valid and reliable instrument for assessing self-directed learning in mainland Chinese nursing students. Nurse educators could use such knowledge to develop their roles and plan to support nursing students in becoming self-directed learners and lifelong learner.


Response of an oscillatory differential delay equation to a periodic stimulus.
Periodic hematological diseases such as cyclical neutropenia or cyclical thrombocytopenia, with their characteristic oscillations of circulating neutrophils or platelets, may pose grave problems for patients. Likewise, periodically administered chemotherapy has the unintended side effect of establishing periodic fluctuations in circulating white cells, red cell precursors and/or platelets. These fluctuations, either spontaneous or induced, often have serious consequences for the patient (e.g. neutropenia, anemia, or thrombocytopenia respectively) which exogenously administered cytokines can partially correct. The question of when and how to administer these drugs is a difficult one for clinicians and not easily answered. In this paper we use a simple model consisting of a delay differential equation with a piecewise linear nonlinearity, that has a periodic solution, to model the effect of a periodic disease or periodic chemotherapy. We then examine the response of this toy model to both single and periodic perturbations, meant to mimic the drug administration, as a function of the drug dose and the duration and frequency of its administration to best determine how to avoid side effects.


Ossification of the laryngeal, tracheal and syringeal cartilages in the domestic fowl.
The process of ossification in the cartilages of the larynx, trachea and syrinx of the domestic fowl has been studied in growing and in adult Golden Comet birds. In the laryngeal cartilages, mineralisation occurred consistently in the body and wings of the cricoid cartilage, in the procricoid cartilage and in the bodies of the arytenoid cartilages. In 7 out of 12 adult birds there were small additional centres in the tips of the rostral processes of the arytenoid cartilages and in one adult bird there were small centres in the caudal processes also. When present, these additional centres were always found bilaterally. Mineralisation in the laryngeal cartilages developed from 105 days post-hatching onwards and was first seen in the bodies of the arytenoid cartilages. Some evidence of a segmental pattern was noted, both in the earliest centres and in the final stage in the caudal region of the body of the tcricoid cartilage. Mineralisation in the trachea developed from 98 days post-hatching onwards. It was first found in the caudal region and spread craniad. The rings at the caudal end of the trachea and the cranial end of the tympanum (but not the last two rings of he tympanum) were always fully involved. Rings in the cranial part of the trachea remained more lightly mineralised. In the syrinx, mineralisation regularly occurred only in the pessulus and in the base of the first bronchial syringeal cartilages. In some birds, it was also encountered more caudally in the cartilages of the primary bronchi. Histological examination showed that, in the early stages, the alizarin staining was due to the presence of mineralised cartilage. At 182 days post-hatching and in the adults, actual bony tissue was observed. The possible significance of the occurrence of ossification in these cartilages of birds in briefly considered.


Whole-chromosome hitchhiking driven by a male-killing endosymbiont.
Neo-sex chromosomes are found in many taxa, but the forces driving their emergence and spread are poorly understood. The female-specific neo-W chromosome of the African monarch (or queen) butterfly Danaus chrysippus presents an intriguing case study because it is restricted to a single 'contact zone' population, involves a putative colour patterning supergene, and co-occurs with infection by the male-killing endosymbiont Spiroplasma. We investigated the origin and evolution of this system using whole genome sequencing. We first identify the 'BC supergene', a broad region of suppressed recombination across nearly half a chromosome, which links two colour patterning loci. Association analysis suggests that the genes yellow and arrow in this region control the forewing colour pattern differences between D. chrysippus subspecies. We then show that the same chromosome has recently formed a neo-W that has spread through the contact zone within approximately 2,200 years. We also assembled the genome of the male-killing Spiroplasma, and find that it shows perfect genealogical congruence with the neo-W, suggesting that the neo-W has hitchhiked to high frequency as the male-killer has spread through the population. The complete absence of female crossing-over in the Lepidoptera causes whole-chromosome hitchhiking of a single neo-W haplotype, carrying a single allele of the BC supergene and dragging multiple non-synonymous mutations to high frequency. This has created a population of infected females that all carry the same recessive colour patterning allele, making the phenotypes of each successive generation highly dependent on uninfected male immigrants. Our findings show how hitchhiking can occur between the physically unlinked genomes of host and endosymbiont, with dramatic consequences.


Teaching Anti-Racism in the Clinical Environment: The Five-Minute Moment for Racial Justice in Healthcare.
Dismantling racism in health care demands that medical education promote racial justice throughout all stages of medical training. However, racial bias can be fostered unintentionally, influencing the way we make decisions as clinicians with downstream effects on patient health and health equity. The development of any anti-racism curriculum in medicine requires the ability to identify racial bias in practices we have not previously recognized as explicitly racist or unjust. This has limited the creation and delivery of effective anti-racism education in health care.


Effect of Preventive and Curative Fingolimod Treatment Regimens on Microglia Activation and Disease Progression in a Rat Model of Multiple Sclerosis.
Fingolimod was the first oral drug approved for multiple sclerosis treatment. Its principal mechanism of action is blocking of lymphocyte trafficking. In addition, recent studies have shown its capability to diminish microglia activation. The effect of preventive and curative fingolimod treatment on the time-course of neuroinflammation was investigated in the experimental autoimmune encephalomyelitis rat model for multiple sclerosis. Neuroinflammatory progression was followed in Dark Agouti female rats after immunization. Positron-Emission tomography (PET) imaging with (R)-[11C]PK11195 was performed on day 11, 15, 19, 27, 29 and 34 during normal disease progression, preventive and curative treatments with fingolimod (1 mg/kg/day). Additionally, bodyweight and clinical symptoms were determined. Preventive treatment diminished bodyweight loss and inhibited the appearance of neurological symptoms. In non-treated rats, PET showed that neuroinflammation peaked in the brainstem at day 19, whereas the imaging signal was decreased in cortical regions. Both preventive and curative treatment reduced neuroinflammation in the brainstem at day 19. Eight days after treatment withdrawal, neuroinflammation had flared-up, especially in cortical regions. Preventive treatment with fingolimod suppressed clinical symptoms and neuroinflammation in the brainstem. After treatment withdrawal, clinical symptoms reappeared together with neuroinflammation in cortical regions, suggesting a different pathway of disease progression.


Using chick forebrain neurons to model neurodegeneration and protection in an undergraduate neuroscience laboratory course.
Since 2009 at Boston College, we have been offering a Research in Neuroscience course using cultured neurons in an in vitro model of stroke. The students work in groups to learn how to perform sterile animal cell culture and run several basic bioassays to assess cell viability. They are then tasked with analyzing the scientific literature in an attempt to identify and predict the intracellular pathways involved in neuronal death, and identify dietary antioxidant compounds that may provide protection based on their known effects in other cells. After each group constructs a hypothesis pertaining to the potential neuroprotection, we purchase one compound per group and the students test their hypotheses using a commonly performed viability assay. The groups generate quantitative data and perform basic statistics on that data to analyze it for statistical significance. Finally, the groups compile their data and other elements of their research experience into a poster for our departmental research celebration at the end of the spring semester.


[Weight-length correlation of newborn infants. A current method for somatic evaluation--standard values].
Over 30,000 deliveries from 1959 to 1980 were evaluated regarding the correlation between birth weight and length of the newborn. The factors influencing the ratios such as sex of the newborn, year of birth and maternal age and parity were investigated. Since 1965, the correlation has no relationship to the year of birth. It was further found that in almost every tenth primigravida the values of the multiparas were not applicable. In primigravidas below age 17 and over age 26 the proportion was 15%. The author therefore proposes that primigravidas are not included into the normal values. About 9,000 neonates of multiparas were used to establish a standard. The correlation is independent of the sex. Male and female newborns can therefore be evaluated by the same standard. A new concept for the evaluation of the somatic state of a newborn was discussed and recommended by the author. This concept uses the usual method which is also used later in life. Evaluation of the body length is correlated to the length of gestation according to length/SSW standards into macrosome , normosome , and microsome and evaluation of the body with a new weight length standard (Figure 10). The advantages are described in several examples. The comparison of the usual evaluation of newborns by weight/SSW with the new concept shows that more differentiated evaluation is possible in about 1/3 of the cases, especially cases with perinatal mortality and RDS. The optimal treatment and observation of the newborn is improved. The scientific evaluation of such problems as fetopathy , premature delivery and dysmature delivery becomes more exact. The danger of misinterpretations and wrong conclusions is diminished.


Landscape genetic structure and evolutionary genetics of insecticide resistance gene mutations in Anopheles sinensis.
Anopheles sinensis is one of the most abundant vectors of malaria and other diseases in Asia. Vector control through the use of insecticides is the front line control method of vector-borne diseases. Pyrethroids are the most commonly used insecticides due to their low toxicity to vertebrates and low repellency. However, the extensive use of insecticides has imposed strong selection pressure on mosquito populations for resistance. High levels of resistance to pyrethroid insecticides and various mutations and haplotypes in the para sodium channel gene that confers knockdown resistance (kdr) have been detected in An. sinensis. Despite the importance of kdr mutations in pyrethroid resistance, the evolutionary origin of the kdr mutations is unknown. This study aims to examine the evolutionary genetics of kdr mutations in relation to spatial population genetic structure of An. sinensis. Adults or larvae of Anopheles sinensis were collected from various geographic locations in China. DNA was extracted from individual mosquitoes. PCR amplification and DNA sequencing of the para-type sodium channel gene were conducted to analyse kdr allele frequency distribution, kdr codon upstream and downstream intron polymorphism, population genetic diversity and kdr codon evolution. The mitochondrial cytochrome c oxidase COI and COII genes were amplified and sequenced to examine population variations, genetic differentiation, spatial population structure, population expansion and gene flow patterns. Three non-synonymous mutations (L1014F, L1014C, and L1014S) were detected at the kdr codon L1014 of para-type sodium channel gene. A patchy distribution of kdr mutation allele frequencies from southern to central China was found. Near fixation of kdr mutation was detected in populations from central China, but no kdr mutations were found in populations from southwestern China. More than eight independent mutation events were detected in the three kdr alleles, and at least one of them evolved multiple times subsequent to their first divergence. Based on sequence analysis of the mitochondrial COI and COII genes, significant and large genetic differentiation was detected between populations from southwestern China and central China. The patchy distribution of kdr mutation frequencies is likely a consequence of geographic isolation in the mosquito populations and the long-term insecticide selection. Our results indicate multiple origins of the kdr insecticide-resistant alleles in An. sinensis from southern and central China. Local selection related to intense and prolonged use of insecticide for agricultural purposes, as well as frequent migrations among populations are likely the explanations for the patchy distribution of kdr mutations in China. On the contrary, the lack of kdr mutations in Yunnan and Sichuan is likely a consequence of genetic isolation and absence of strong selection pressure. The present study compares the genetic patterns revealed by a functional gene with a neutral marker and demonstrates the combined impact of demographic and selection factors on population structure.


The response of thyrotropin and triiodothyronine to various doses of thyrotropin releasing hormone in normal man.
The relationship between serum thyrotropin (TSH) and serum triiodothyronine (T3) before and after injection of different doses of thyrotropin releasing hormone (TRH), given as single injections or as multiple injections with short intervals, was investigated in normal men. A positive correlation between prestimulated serum TSH and serum T3 levels and between the increase in the serum TSH and the serum T3 levels after TRH was found when repeated tests were performed in the same individual. There was a dose dependent TSH and T3 response to TRH. The smallest dose that produced a maximal response of both TSH and T3 was only 30 mug TRH. After six injections of 30 mug TRH with an interval of 30 minutes the increase in TSH was two times and the increase in T3 was three times as high as the maximal increase after single injections of TRH. This test with multiple injections of TRH may prove to be of clinical value in the measurement of both pituitary and thyroid function in selected patients. The close positive correlation between the serum TSH and serum T3 levels in basal conditions, demonstrated in four normal subjects in this study, probably reflects the steady state level determined by the hypothalamus from which the feedback control of TSH secretion operates.


ALS-linked FUS mutations confer loss and gain of function in the nucleus by promoting excessive formation of dysfunctional paraspeckles.
Mutations in the FUS gene cause amyotrophic lateral sclerosis (ALS-FUS). Mutant FUS is known to confer cytoplasmic gain of function but its effects in the nucleus are less understood. FUS is an essential component of paraspeckles, subnuclear bodies assembled on a lncRNA NEAT1. Paraspeckles may play a protective role specifically in degenerating spinal motor neurons. However it is still unknown how endogenous levels of mutant FUS would affect NEAT1/paraspeckles. Using novel cell lines with the FUS gene modified by CRISPR/Cas9 and human patient fibroblasts, we found that endogenous levels of mutant FUS cause accumulation of NEAT1 isoforms and paraspeckles. However, despite only mild cytoplasmic mislocalisation of FUS, paraspeckle integrity is compromised in these cells, as confirmed by reduced interaction of mutant FUS with core paraspeckle proteins NONO and SFPQ and increased NEAT1 extractability. This results in NEAT1 localisation outside paraspeckles, especially prominent under conditions of paraspeckle-inducing stress. Consistently, paraspeckle-dependent microRNA production, a readout for functionality of paraspeckles, is impaired in cells expressing mutant FUS. In line with the cellular data, we observed paraspeckle hyper-assembly in spinal neurons of ALS-FUS patients. Therefore, despite largely preserving its nuclear localisation, mutant FUS leads to loss (dysfunctional paraspeckles) and gain (excess of free NEAT1) of function in the nucleus. Perturbed fine structure and functionality of paraspeckles accompanied by accumulation of non-paraspeckle NEAT1 may contribute to the disease severity in ALS-FUS.


Bendamustine for the treatment of multiple myeloma in first-line and relapsed-refractory settings: a review of clinical trial data.
Multiple myeloma (MM) is a hematologic malignancy characterized by abnormal growth and/or dysregulation of plasma cells leading to the build-up of malignant plasma cells in the bone marrow and increased production of monoclonal immunoglobulins. Treatment modalities for MM include autologous stem cell transplant (ASCT), chemotherapy with conventional and immunomodulatory agents, radiation therapy and adjunct therapies. Bendamustine is a synthetic chemotherapeutic agent combining the alkylating properties of a mustard group with the activities of a benzimidazole ring, giving it a unique alkylating activity compared with other alkylating agents. Bendamustine has proven activity in both newly diagnosed and relapsed-refractory MM. Bendamustine has also demonstrated activity in MM after relapse from ASCT, and has recently been used successfully as a conditioning regimen for ASCT in combination with melphalan. Bendamustine is generally well tolerated, with the majority of adverse events being due to bone marrow suppression. Extramedullary toxicity is infrequent and usually mild.


How to demonstrate the reversibility of end-organ function before implantation of left ventricular assist device in INTERMACS profile 2 patients?
For the time being, in Japan, two recently approved implantable ventricular assist devices (VADs) are indicated only when a patient has been listed for heart transplantation or approved to be eligible for heart transplantation by in-hospital committee. The reversibility of end-organ dysfunction must be expected before VAD implantation, but it is often hard to prove during worsening clinical status. We report two patients whose end-organ dysfunction had been eventually demonstrated to be reversible by invasive procedures such as transluminal liver biopsy or transient insertion of intra-aortic balloon pumping.


Agenesis of the vermiform appendix.
Agenesis of the vermiform appendix is very rare. The incidence is estimated to be one in 100,000 laparotomies for suspected appendicitis. Several criteria have to be met before the investigator can conclude that the appendix is congenitally absent. This case is reported to bring this entity to the attention of surgeons who may encounter a similar situation during celioscopy. A 29-year-old patient was admitted through the emergency room with the chief complaint of abdominal pain. Acute appendicitis was suspected, and he was accordingly prepared for celioscopy. This report presents a patient with vermiform appendix agenesis diagnosed at celioscopy with concomitant mesenteric lymphadenitis. Agenesis of the vermiform appendix is very rare, and the diagnosis should not be made unless the ileocecal and retrocecal area are thoroughly explored.


Binding of human interferon alpha to cells of different sensitivities: studies with internally radiolabeled interferon retaining full biological activity.
The characteristics of interferon binding to various cells with different interferon sensitivity were studied by using [3H]leucine-labeled, pure human interferon alpha from Namalwa cells. Scatchard analysis of the binding data on cells sensitive to interferon alpha (human FL and fibroblasts and bovine MDBK) indicated the presence of two kinds of binding sites with high and low affinities. The binding constants of the high-affinity sites in these cells were similar (4 X 10(10) to 11 X 10(10) M-1). Cells insensitive to human interferon alpha (human HEC-1 and mouse L cells) were shown to have only low-affinity sites, suggesting that high-affinity binding sites are indispensable for interferon sensitivity and represent interferon receptors. However, the number of sites in three human diploid fibroblast strains and one strain trisomic for chromosome 21 were not proportionally correlated to the interferon sensitivity of the cells. The high-affinity binding to human cells was completely inhibited by both nonradioactive human interferons alpha and beta in a similar manner, but binding to bovine MDBK cells, on which human interferon beta is practically inactive, was inhibited effectively only by interferon alpha and not by beta. These results suggest that the receptor for human interferon alpha is common to human interferon beta in human cells, whereas the receptor on bovine cells binds only human interferon alpha.


Health related practices of students of the University of Ibadan.
University students are predisposed by environmental factors to adoption of habits that impact on their health Smoking, alcohol abuse, a sedentary lifestyle and failure to use health services appropriately may all impair current or future health. These students represent a large population of young persons but have not been recognised as a specific concern among health care planners except in the area of reproductive health and sexuality. Seven hundred and fifty students of the University of Ibadan were randomly selected from the halls of residence by multi stage sampling. Verbal consent was obtained and trained student peers administered questionnaires to the subjects Seven hundred and twenty-four students returned completed questionnaires, an overall response rate of 96.5% The respondents consisted of 484 males and 240 females Six hundred and two were undergraduate and 122 were graduate students. All the university's faculties were represented. Most (92.3%) of the respondents were single The mean age of the group was 23.8 years (SD 4.6). Current and ever used smoking prevalence was 5.7% and 23.2% respectively. Smokers were predominantly males pursuing degrees in the Arts and Humanities. Current and ever use of alcohol were reported by 24.7% and 46.1% respectively of the students. Main associations with alcohol consumption were with male gender, age less than 30 years, single marital status and non-medical course of study. Only 12 students (1.8%) exercise daily. This study demonstrates the need for continued surveillance of health related habits of young persons, not only for the benefit of the individual students but also for the general population for whom these educated persons represent lifestyle models.


TOR signaling is required for amino acid stimulation of early trypsin protein synthesis in the midgut of Aedes aegypti mosquitoes.
Blood meal digestion in mosquitoes occurs in two phases, an early phase that is translationally regulated, and a late phase that is transcriptionally regulated. Early trypsin is a well-characterized serine endoprotease that is representative of other early phase proteases in the midgut that are only synthesized after feeding. Since the kinase Target of Rapamycin (TOR) has been implicated as a nutrient sensor in other systems, including the mosquito fat body, we tested if TOR signaling is involved in early trypsin protein synthesis in the mosquito midgut in response to feeding. We found that ingestion of an amino acid meal by female mosquitoes induces early trypsin protein synthesis, coincident with phosphorylation of two known TOR target proteins, p70S6 kinase (S6K) and the translational repressor 4E-Binding Protein (4E-BP). Moreover, in vitro culturing of midguts from unfed mosquitoes led to amino acid-dependent phosphorylation of S6K and 4E-BP which could be blocked by treatment with rapamycin, a TOR-specific inhibitor. Lastly, by injecting mosquitoes with TOR double stranded RNA (dsRNA) or rapamycin, we demonstrated that TOR signaling was required in vivo for both phosphorylation of S6K and 4E-BP in the midgut, and for translation of early trypsin mRNA in response to amino acid feeding. It may be possible to target the TOR signaling pathway in the midgut to inhibit blood meal digestion, and thereby, decrease fecundity and the spread of mosquito borne diseases.