Automatic Diagnosis of COVID-19 Pneumonia using Artificial Intelligence Deep Learning Algorithm Based on Lung Computed Tomography Images.
The lung computed tomography (CT) scan contains valuable information and patterns that provide the possibility of early diagnosis of COVID-19 disease as a global pandemic by the image processing software. In this research, based on deep learning of artificial intelligence, the software has been designed that is used clinically to diagnose COVID-19 disease with high accuracy. Convolutional neural network architecture developed based on Inception-V3 for deep learning of lung image patterns, feature extraction, and image classification. The theory of transfer learning was utilized to increase the learning power of the system. Changes applied in the network layers to increase the detection power. The process of learning was repeated 30 times. All diagnostic statistical parameters of the diagnostic were analyzed to validate the software. Based on the data of Imam Khomeini Hospital in Sari, the validity, sensitivity, and accuracy of the software in diagnosing of affected to COVID-19 and nonaffected to it were obtained 98%, 98%, and 98%, respectively. Diagnostic statistical parameters on some data were 100%. The modified algorithm of Inception-V3 applied to heterogeneous data also had acceptable precision. The proposed basic architecture of Inception-v3 utilized for this research has an admissible speed and exactness in learning CT scan images of patients' lungs, and diagnosis of COVID-19 pneumonia, which can be utilized clinically as a powerful diagnostic tool.


Abnormal predefecatory colonic motor patterns define constipation in obstructed defecation.
The pathophysiology of constipation in the syndrome of obstructed defecation is unknown. Using 24-hour pancolonic manometric recordings of the unprepared colon to record basal pressures and spontaneous defecation episodes, we tested the hypothesis that the frequency, timing, or spatial distribution of propagating colonic pressure waves is abnormal in patients with obstructed defecation. In 11 patients with obstructed defecation and 16 healthy controls, pressures were recorded using a nasocolonic catheter that was positioned such that 16 recording sites spanned the unprepared colon at 7.5-cm intervals. The overall frequency of propagating sequences (PS) in the colon did not differ between patients and controls. When compared with controls, patients had a significant increase in the frequency of retrograde and antegrade PS (P < 0.05) in the left colon and a significant reduction in the amplitude of propagating pressure waves throughout the entire colon (P < 0.03). Defecation occurred in 6 of 11 patients and 9 of 16 controls. In the 15 minutes before defecation, controls showed a highly significant increase in frequency (P = 0.001) and amplitude (P = 0.01) of PS. In contrast, patients did not demonstrate this or the typical spatiotemporal organization of PS normally observed before expulsion of stool. Patients with obstructed defecation lack the normal predefecatory augmentation in frequency and amplitude of propagating pressure waves and lack the normal stereotypic spatiotemporal patterning of colonic pressure waves that would normally culminate in effective expulsion of stool.


Evolution of the oxygen sensitivity of cytochrome c oxidase subunit 4.
Vertebrates possess two paralogs of cytochrome c oxidase (COX) subunit 4: a ubiquitous COX4-1 and a hypoxia-linked COX4-2. Mammalian COX4-2 is thought to have a role in relation to fine-tuning metabolism in low oxygen levels, conferred through both structural differences in the subunit protein structure and regulatory differences in the gene. We sought to elucidate the pervasiveness of this feature across vertebrates. The ratio of COX4-2/4-1 mRNA is generally low in mammals, but this ratio was higher in fish and reptiles, particularly turtles. The COX4-2 gene appeared unresponsive to low oxygen in nonmammalian models (zebrafish, goldfish, tilapia, anoles, and turtles) and fish cell lines. Reporter genes constructed from the amphibian and reptile homologues of the mammalian oxygen-responsive elements and hypoxia-responsive elements did not respond to low oxygen. Unlike the rodent ortholog, the promoter of goldfish COX4-2 did not respond to hypoxia or anoxia. The protein sequences of the COX4-2 peptide showed that the disulfide bridge seen in human and rodent orthologs would be precluded in other mammalian lineages and lower vertebrates, all of which lack the requisite pair of cysteines. The coordinating ligands of the ATP-binding site are largely conserved across mammals and reptiles, but in Xenopus and fish, sequence variations may disrupt the ability of the protein to bind ATP at this site. Collectively, these results suggest that many of the genetic and structural features of COX4-2 that impart responsiveness and benefits in hypoxia may be restricted to the Euarchontoglires lineage that includes primates, lagomorphs, and rodents.


Mono- and dinuclear cobalt complexes with chelating or bridging bidentate P,N phosphino- and phosphinito-oxazoline ligands: synthesis, structures and catalytic ethylene oligomerisation.
Cobalt(ii) complexes of the type [CoCl(2)(P,N)], where P,N represents a heterobidentate phosphino- or phosphinito-oxazoline-type ligand, have been synthesised and characterised by infrared spectroscopy and elemental analysis. Their molecular structures were established by single-crystal X-ray diffraction in the solid state. Whereas the phosphino-oxazoline complex [CoCl(2){Ph(2)PCH(2)ox(Me2)}] (Ph(2)PCH(2)ox(Me2) = 2-[(diphenylphosphanyl)-methyl]-4,4-dimethyl-4,5-dihydro-oxazole) () and the phosphinito-oxazoline complexes [CoCl(2){Ph(2)POCH(2)ox(Me2)}] (Ph(2)POCH(2)ox(Me2) = 1-[4,4-dimethyl-2{1-oxy(diphenylphosphino)-1-methyl}]-4,5-dihydro-oxazole) () and [CoCl(2){Ph(2)POCMe(2)ox(Me2)}] (Ph(2)POCMe(2)ox(Me2) = 1-[4,4-dimethyl-2- [1-oxy(diphenylphosphino)-1-methylethyl]]-4,5-dihydrooxazole) () are mononuclear, the phosphino-oxazoline complexes [CoCl(2){micro-i-Pr(2)PCH(2)ox}](2) (i-Pr(2)PCH(2)ox = 2-[(diisopropyl-phosphanyl)-methyl]-4,5-dihydro-oxazole) () and [CoCl(2){micro-Ph(2)PCH(2)ox}](2) (Ph(2)PCH(2)ox = 2-[(diphenyl-phosphanyl)-methyl]-4,5-dihydro-oxazole) () are dinuclear compounds and contain two bridging phosphino-oxazoline ligands which form a 10-membered ring. In the course of this work, the zwitterionic complex [CoCl(3){Ph(2)PCH(2)C(O)OCH(2)CMe(2)NH(3)] () was obtained and characterised by X-ray diffraction in which the oxazoline ring has been opened. Air-oxidation of the phosphine function of the mononuclear P,N chelate complex yielded the blue N,O-bridged, centrosymmetric dinuclear complex [[upper bond 1 start]CoCl(2){micro-OPPh(2)CH(2)[lower bond 1 start]C[double bond, length as m-dash]N[upper bond 1 end]CMe(2)CH(2)O[lower bond 1 end]}](2) () which contains a 12-membered ring. All these complexes are paramagnetic and their magnetic moments in solution were measured by the Evans method. Complexes were evaluated in the catalytic oligomerisation of ethylene with AlEtCl(2) or methylaluminoxane (MAO) as cocatalysts and provided moderate activities. In the presence of AlEtCl(2) (6-14 equiv.), the selectivities for ethylene dimers were higher than 92% and complex showed the highest turnover frequency with 14 equiv. of AlEtCl(2). When MAO was used as cocatalyst, the catalytic activities were similar to those with AlEtCl(2) but significant amounts of C(6)-C(12) oligomers were produced.


Metabolic profiling for detection of Staphylococcus aureus infection and antibiotic resistance.
Due to slow diagnostics, physicians must optimize antibiotic therapies based on clinical evaluation of patients without specific information on causative bacteria. We have investigated metabolomic analysis of blood for the detection of acute bacterial infection and early differentiation between ineffective and effective antibiotic treatment. A vital and timely therapeutic difficulty was thereby addressed: the ability to rapidly detect treatment failures because of antibiotic-resistant bacteria. Methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) were used in vitro and for infecting mice, while natural MSSA infection was studied in humans. Samples of bacterial growth media, the blood of infected mice and of humans were analyzed with combined Gas Chromatography/Mass Spectrometry. Multivariate data analysis was used to reveal the metabolic profiles of infection and the responses to different antibiotic treatments. In vitro experiments resulted in the detection of 256 putative metabolites and mice infection experiments resulted in the detection of 474 putative metabolites. Importantly, ineffective and effective antibiotic treatments were differentiated already two hours after treatment start in both experimental systems. That is, the ineffective treatment of MRSA using cloxacillin and untreated controls produced one metabolic profile while all effective treatment combinations using cloxacillin or vancomycin for MSSA or MRSA produced another profile. For further evaluation of the concept, blood samples of humans admitted to intensive care with severe sepsis were analyzed. One hundred thirty-three putative metabolites differentiated severe MSSA sepsis (n = 6) from severe Escherichia coli sepsis (n = 10) and identified treatment responses over time. Combined analysis of human, in vitro, and mice samples identified 25 metabolites indicative of effective treatment of S. aureus sepsis. Taken together, this study provides a proof of concept of the utility of analyzing metabolite patterns in blood for early differentiation between ineffective and effective antibiotic treatment in acute S. aureus infections.


Intravenous immunoglobulin and autoimmune disease.
Intravenous immunoglobulin (IVIg) has been found to have a multitude of uses. However, IVIg is a pooled blood product and as a result a limited resource. At replacement doses (200-400 mg/kg/month) it is of critical utility in the treatment of primary and secondary antibody deficiencies. High-dose immunoglobulin (hdIVIg) given at doses of up to 2 g/kg/day has immunomodulatory action mediated via a number of different effects. First used in the 1980s for the treatment of idiopathic thrombocytopenic purpura, hdIVIg has found uses in a wide range of autoimmune conditions, though for many conditions the evidence base lacks formal randomized controlled trials (RCTs). This article will detail the issues regarding the manufacture and clinical aspects of administration of hdIVIg and its uses, especially with regards to the treatment of autoimmune disease.


Dental Rehabilitation for Free Fibula Flap-Reconstructed Mandible with Scar Contracture: A Technical Note.
Dental rehabilitation with osseointegrated implants in reconstructed mandibles is a common procedure, but the technique still requires improvement, especially in its reliability and technical simplification. We herein report dental rehabilitation of a free fibula-reconstructed mandible with scar contracture. A vestibuloplasty technique with application of a polyglycolic acid (PGA) sheet is described. The implants were inserted into a viable fibula flap with severe scar contracture of the overlying epithelium resulting from vascular instability in skin paddle. Only the fibula periosteum was sutured after implant insertion; exposed surfaces were covered with a combination of PGA sheet and fibrin sealant. The area with PGA sheet coverage gradually healed with moderate contracture. The epithelium around the almost implants became immobilized. The implant-supported removable partial denture with custom titanium bar was acceptable. Dental rehabilitation is possible for reconstructed mandibles with severe scar contracture. Application of a PGA sheet may be useful for vestibuloplasty in patients with reconstructed mandibles.


A 2H NMR relaxation experiment for the measurement of the time scale of methyl side-chain dynamics in large proteins.
An NMR experiment is presented for the measurement of the time scale of methyl side-chain dynamics in proteins that are labeled with methyl groups of the (13)CHD(2) variety. The measurement is accomplished by selecting a magnetization mode that to excellent approximation relaxes in a single-exponential manner with a T(1)-like rate. The combination of R(1)((13)CHD(2)) and R(2)((13)CHD(2)) (2)H relaxation rates facilitates the extraction of motional parameters from (13)CHD(2)-labeled proteins exclusively. The utility of the methodology is demonstrated with applications to proteins with tumbling times ranging from 2 ns (protein L, 7.5 kDa, 45 degrees C) to 54 ns (malate synthase G, 82 kDa, 37 degrees C); dynamics parameters are shown to be in excellent agreement with those obtained in (2)H NMR studies of other methyl isotopomers. A consistency relationship is found to exist between R(1)((13)CHD(2)) and the relaxation rates of pure longitudinal and quadrupolar order modes in (13)CH(2)D-labeled methyl groups, and experimental rates measured for a number of proteins are shown to be in excellent agreement with expectations based on theory. The present methodology extends the applicability of (2)H relaxation methods for the quantification of side-chain dynamics in high molecular weight proteins.


Opposing actions of chronic Delta9-tetrahydrocannabinol and cannabinoid antagonists on hippocampal long-term potentiation.
Memory deficits produced by marijuana arise partly via interaction of the psychoactive component, Delta(9)-tetrahydrocannabinol (Delta(9)-THC), with cannabinoid receptors in the hippocampus. Although cannabinoids acutely reduce glutamate release and block hippocampal long-term potentiation (LTP), a potential substrate for learning and memory, the consequences of prolonged exposure to Delta(9)-THC for hippocampal function are poorly understood. Rats were injected with Delta(9)-THC (10 mg/kg, i.p., q.d.) for 1, 3, or 7 d, and electrophysiological recordings were performed in hippocampal slices 1d after the final injection. At this time, Delta(9)-THC was undetectable in hippocampus using liquid chromatography-mass spectrometry (LC-MS). Hippocampal LTP generated using high-frequency (HFS) or theta burst stimulation was not observed in brain slices from the 7-d Delta(9)-THC-treated animals. Delta(9)-THC also blocked HFS-LTP after 3 d, but not 1 d of treatment. The complete blockade of LTP persisted for 3 d after the last Delta(9)-THC injection, and full reversal of the LTP deficit was not observed up to 14 d following Delta(9)-THC withdrawal. The cannabinoid antagonist AM251 (2 mg/kg), administered before each Delta(9)-THC injection prevented the blockade of LTP, and 7-d treatment with AM251 alone significantly increased the level of LTP. Chronic Delta(9)-THC also produced tolerance to the inhibition of synaptic GABA, but not glutamate release by the agonist WIN55,212-2. These data define consequences of repeated Delta(9)-THC exposure for synaptic plasticity in the hippocampus that may help explain memory impairments in humans following chronic marijuana use.


Efficacy of fosaprepitant combined with tropisetron plus dexamethasone in preventing nausea and emesis during fractionated radiotherapy with weekly cisplatin chemotherapy: interim analysis of a randomized, prospective, clinical trial using competing risk analysis.
There are no well-recognized guidelines for antiemesis during concurrent chemoradiotherapy (CCRT) for cervical cancer (CC) and nasopharyngeal cancer (NPC) until now. The study was designed to assess the efficacy and safety of fosaprepitant combined with tropisetron and dexamethasone in preventing nausea and vomiting during 5 weeks of fractionated radiotherapy and concomitant weekly low-dose cisplatin chemotherapy in patients with CC or NPC. Patients with CC or NPC were scheduled to receive fractionated radiotherapy and weekly cisplatin (25-40 mg/m2) chemotherapy for at least 5 weeks. Patients stratified by tumor type and induction chemotherapy were 1:1 randomly assigned to receive fosaprepitant, tropisetron, and dexamethasone or tropisetron plus dexamethasone as an antiemetic regimen. Efficacy was assessed primarily by the cumulative incidence of emesis after 5 weeks of treatment, and safety by adverse events (AEs). Between July 2020 and July 2022, 116 patients consented to the study of whom 103 were included in this interim analysis (fosaprepitant group [N = 52] vs control group [N = 51]). The cumulative incidence of emesis at 5 weeks (competing risk analysis) was 25% (95% CI 14.2-37.4) for the fosaprepitant group compared with 59% (95% CI 43.9-71.0) for the control group. There was a significantly lower cumulative risk of emesis in the fosaprepitant group (HR 0.35 [95% CI 0.19-0.64]; p < 0.001). Fosaprepitant was well tolerated as the incidences of adverse events in the two groups were comparable. The addition of fosaprepitant to tropisetron plus dexamethasone significantly reduced the risk of nausea and vomiting during 5 weeks of CCRT in patients with CC or NPC, and fosaprepitant was well tolerated. The trial was registered with ClinicalTrials.gov on October 3, 2022, number NCT05564286.


[Myorhythmia emerging on day 15 in a case of an infarction in the midbrain-pontine tegmentum].
A 72-year-old woman was admitted to our hospital because of dysarthria, dysstasia, and diplopia. Examination demonstrated wall-eyed bilateral internuclear ophthalmoplegia (WEBINO) syndrome, dysarthria, truncal ataxia, and dysmetria of the four limbs. Cranial MR images demonstrated an acute infarct in the paramedian, lower midbrain-pontine tegmentum. Peduncular hallucinosis was observed during the first several days of hospitalization. Myorhythmia (skeletal myoclonus) appeared as early as day 15 and has persisted to date, with a frequency of approximately 1.5 Hz on the head and chin, and 3.5 Hz on the left upper extremity. Cranial MR images 5 months after onset demonstrated bilateral inferior olivary hypertrophy, while palatal tremor appeared later. Only a limited number of case reports are available in which myorhythmia emerged as early as in our case after the onset of a brain infarct.


On training targets for deep learning approaches to clean speech magnitude spectrum estimation.
Estimation of the clean speech short-time magnitude spectrum (MS) is key for speech enhancement and separation. Moreover, an automatic speech recognition (ASR) system that employs a front-end relies on clean speech MS estimation to remain robust. Training targets for deep learning approaches to clean speech MS estimation fall into three categories: computational auditory scene analysis (CASA), MS, and minimum mean square error (MMSE) estimator training targets. The choice of the training target can have a significant impact on speech enhancement/separation and robust ASR performance. Motivated by this, the training target that produces enhanced/separated speech at the highest quality and intelligibility and that which is best for an ASR front-end is found. Three different deep neural network (DNN) types and two datasets, which include real-world nonstationary and coloured noise sources at multiple signal-to-noise ratio (SNR) levels, were used for evaluation. Ten objective measures were employed, including the word error rate of the Deep Speech ASR system. It is found that training targets that estimate the a priori SNR for MMSE estimators produce the highest objective quality scores. Moreover, it is established that the gain of MMSE estimators and the ideal amplitude mask produce the highest objective intelligibility scores and are most suitable for an ASR front-end.


The control by respiration of the uptake of alpha-methyl glucoside in Escherichia coli K12.
The uptake of methyl alpha-D-glucopyranoside (alpha-MG) by Escherichia coli K12 was decreased by the addition of substrates which stimulated the rate of oxygen consumption by the cells. The inhibition, which occurred only at non-saturating concentrations of alpha-MG, was not the result of a stimulation of the rate of exit of intracellular alpha-MG, and was abolished by the presence of carbonyl cyanide m-chlorophenylhydrazone or sodium azide. Since those drugs inhibit energy conservation at the respiratory chain and did not alter significantly the rate of oxygen consumption under the conditions for the assay of alpha-MG uptake, it appears that the inhibition of the transport system by respirable substrates is mediated by some form of energy derived from respiration.


Multimodal fMRI tractography in normal subjects and in clinically recovered traumatic brain injury patients.
In this study, we defined an operator-independent protocol for reconstructing the anatomical connections originating from fMRI activations in order to demonstrate that results obtained with this protocol are affected by alterations of functional activations. Seven healthy volunteers and two patients who sustained traumatic brain injury underwent an fMRI with a finger tapping task and a DTI scan. Cortical fMRI activations were used directly as seed mask for tractography for the reconstruction of individual motor pathways. On patients we observed a different motor network if compared to healthy subjects. However, when the activations of healthy subjects were used as seed masks for the tractography in patients, we observed for the patients a pattern of connectivity more similar to what was observed for healthy subjects. At the same time, when the activations of patients were used for the tractography on healthy subjects, we obtained patterns of connectivity similar to those obtained for patients. These results show the potential of the integration of fMRI and tractography for clarifying the mechanisms of cortical plasticity in the recovery of motor functions.


Use of simple clinical and laboratory predictors to differentiate influenza from dengue and other febrile illnesses in the emergency room.
Clinical differentiation of influenza from dengue and other febrile illnesses (OFI) is difficult, and available rapid diagnostic tests have limited sensitivity. We conducted a retrospective study to compare clinical and laboratory findings between (i) influenza and dengue and (ii) influenza and OFI. Of 849 enrolled patients, the mean time between illness onset and hospital presentation was 1.7, 3.7, and 3 days for influenza, dengue, and OFI, respectively. Among pediatric patients (≤18 years) (445 influenza, 24 dengue, and 130 OFI), we identified absence of rashes, no leukopenia, and no marked thrombocytopenia (platelet counts <100 × 10(9) cells/L) as predictors to distinguish influenza from dengue, whereas rhinorrhea, malaise, sore throat, and mild thrombocytopenia (platelet counts 100-149 × 10(9)/L) were predictors that differentiated influenza from OFI. Among adults (>18 years) (81 influenza, 124 dengue, and 45 OFI), no leukopenia and no marked thrombocytopenia distinguished influenza from dengue, while rhinorrhea and malaise differentiated influenza from OFI. A diagnostic algorithm developed to distinguish influenza from dengue using rash, leukopenia, and marked thrombocytopenia showed >90% sensitivity to identify influenza in pediatric patients. This study identified simple clinical and laboratory parameters that can assist clinicians to distinguish influenza from dengue and OFI. These findings may help clinicians diagnose influenza and facilitate appropriate management of affected patients, particularly in resource-poor settings.


Dual-Modal Therapeutic Role of the Lactate Oxidase-Embedded Hierarchical Porous Zeolitic Imidazolate Framework as a Nanocatalyst for Effective Tumor Suppression.
The increasing evidence supports the fact that lactate in the tumor microenvironment (TME) plays a vital role in tumor proliferation, metastasis, and recurrence, which in turn is emerging as one of the most interesting molecular targets for tumor treatment. Here, hierarchical porous zeolitic imidazolate framework-8 (ZIF-8) as the nanocarrier is fabricated to simultaneously load lactate oxidase (LOD) and Fe3O4 nanoparticles (NPs), called LOD & Fe3O4@ZIF-8 NPs (LFZ NPs), for tumor therapy. On one hand, the sharp consumption of lactate in the TME by LOD will change the essential "soil" where tumor cells live so as to suppress tumor rapid growth. On the other hand, hydrogen peroxide (H2O2) is produced in the TME from the oxidation of lactate catalyzed by LOD and subsequently converted to highly toxic hydroxyl radicals (•OH) catalyzed by Fe3O4 NPs via Fenton-like reactions to kill tumor cells. Based on the endogenous catalysis, this dual-modal strategy of tumor therapy based on lactate is simple, safe, and effective, which deserves to be well concerned.


Improved binding affinity and interesting selectivities of aminopyrimidine-bearing carbohydrate receptors in comparison with their aminopyridine analogues.
Due to the problems with the exact prediction of the binding properties of an artificial carbohydrate receptor, the identification of characteristic structural features, having the ability to influence the binding properties in a predictable way, is of high importance. The purpose of our investigation was to examine whether the previously observed higher affinity of 2-aminopyrimidine-bearing carbohydrate receptors in comparison with aminopyridine substituted analogues represents a general tendency of aminopyrimidine-bearing compounds. Systematic binding studies on new compounds consisting of 2-aminopyrimidine groups confirmed such a tendency and allowed the identification of interesting structure-activity relationships. Receptors having different symmetries showed systematic preferences for specific glycosides, which are remarkable for such simple receptor systems. Particularly suitable receptor architectures for the recognition of selected glycosides were identified and represent a valuable base for further developments in this field.


Higher incidence of new vertebral fractures following percutaneous vertebroplasty and kyphoplasty--fact or fiction?
New vertebral fractures after percutaneous vertebroplasty or kyphoplasty are said to result from biomechanical changes induced by cementation. Fact or fiction? The reported incidences for new vertebral fractures after cementation or after conservative therapy vary widely. This is mainly due to differences in their design, more specifically as to the duration of followup. Therefore a systematic review of the literature was performed, searching for comparable publications to assess the potential risk of new vertebral fractures following vertebroplasty and kyphoplasty versus conservative treatment. Studies were only included if they granted a standardized one-year radiological follow-up, so improving comparability. However, a high degree of heterogeneity was still seen among the results, which made it impossible to state that cement augmentation is as safe as conservative treatment with respect to new fractures. In other words, it was impossible to separate facts from fiction with the studies available to-day. The combined odds ratio of vertebroplasty and kyphoplasty versus conservative treatment, namely 0.96, gave a hint that there might be little difference. Large scale randomized studies will be necessary.


Genome-wide identification, functional prediction, and evolutionary analysis of the R2R3-MYB superfamily in Brassica napus.
R2R3-MYB transcription factors (TFs) have been shown to play important roles in plants, including in development and in various stress conditions. Phylogenetic analysis showed the presence of 249 R2R3-MYB TFs in Brassica napus, called BnaR2R3-MYB TFs, clustered into 38 clades. BnaR2R3-MYB TFs were distributed on 19 chromosomes of B. napus. Sixteen gene clusters were identified. BnaR2R3-MYB TFs were characterized by motif prediction, gene structure analysis, and gene ontology. Evolutionary analysis revealed that BnaR2R3-MYB TFs are mainly formed as a result of whole-genome duplication. Orthologs and paralogs of BnaR2R3-MYB TFs were identified in B. napus, B. rapa, B. oleracea, and Arabidopsis thaliana using synteny-based methods. Purifying selection was pervasive within R2R3-MYB TFs. Kn/Ks values lower than 0.3 indicated that BnaR2R3-MYB TFs are being functionally converged. The role of gene conversion in the formation of BnaR2R3-MYB TFs was significant. Cis-regulatory elements in the upstream regions of BnaR2R3-MYB genes, miRNA targeting BnaR2R3MYB TFs, and post translational modifications were identified. Digital expression data revealed that BnaR2R3-MYB genes were highly expressed in the roots and under high salinity treatment after 24 h. BnaMYB21, BnaMYB141, and BnaMYB148 have been suggested for improving salt-tolerant B. napus. BnaR2R3-MYB genes were mostly up regulated on the 14th day post inoculation with Leptosphaeria biglobosa and L. maculan. BnaMYB150 is a candidate for increased tolerance to Leptospheria in B. napus.


Applying Public Health Frameworks to Advance the Promotion of Mental Health Among Asian American Children.
Asian American (ASA) children experience high rates of mental health problems. Although there is a pressing need to utilize population approaches, emerging frameworks from the fields of public and population health have not been applied to ASA children. This paper addresses this gap by first discussing applications of the National Prevention Strategy (NPS), a population strategy developed from the Social Determinants of Health perspective, to guide ASA prevention work. Next, we provide a practical example to illustrate how the NPS can be applied to prevention program design (using ParentCorps as an example) and dissemination and implementation processes to broadly address ASA children's mental health needs. Finally, we present preliminary data on the feasibility of applying this population strategy to ASA families and a framework for researchers who are considering disseminating and implementing evidence-based programs to ASA or ethnic minority pediatric populations.


Interaction of HIV-1 ribonuclease H with polypurine tract containing RNA-DNA hybrids.
The mode of action of the RNase H activity from HIV-1 was analyzed with a purified recombinant p66/p51 reverse transcriptase RT/RNase H protein and RNA-DNA hybrid consisting of RNA harboring the polypurine tract (ppt) and three complementary synthetic DNA oligonucleotides. Upon incubation of this preformed RNA-DNA hybrid with the p66/p51 RT/RNase H, a cleavage pattern is observed that indicates endonucleolytic RNase H activity with some sequence specificity for the next to last nucleotide of the 3'-end of the ppt RNA and one cut within the ppt. The RNase H avoids cleavage of G or A stretches. During RNA-directed DNA synthesis the RNA is hydrolyzed in a concerted action of RT and RNase H whereby the RNase H exhibits endonuclease as well as 3'-5'-exonuclease activity. The distance between the active centers of the RT and RNase H corresponds to 18 base pairs of the RNA-DNA hybrid. Plus-strand DNA-directed DNA synthesis initiates exactly at the next to last nucleotide of the 3'-end of the ppt RNA by means of the RNase H activity.


Clozapine-induced seizures and EEG changes.
Clozapine, an atypical antipsychotic drug, is widely used in refractory schizophrenia. At New Hampshire Hospital, 7 of the first 35 patients treated (20%) had convulsions. Patterns were tonic-clonic (5), complex partial (2), and myoclonic (3). Seizures were dose-related and may be anticholinergic in etiology. EEG changes are frequent with clozapine, particularly as dosage is increased. Twenty-six of 35 patients (74%) had EEG abnormalities at some time during clozapine treatment. EEG is a sensitive means of detecting clinical toxicity. When EEG abnormalities (slowing, dysrhythmia, or paroxysmal discharges) are detected, immediately lowering the dose by at least 25-50 mg per day and adjusting weekly until EEG returns to baseline can reduce the incidence of seizures.


AnABlast: a new in silico strategy for the genome-wide search of novel genes and fossil regions.
Genome annotation, assisted by computer programs, is one of the great advances in modern biology. Nevertheless, the in silico identification of small and complex coding sequences is still challenging. We observed that amino acid sequences inferred from coding-but rarely from non-coding-DNA sequences accumulated alignments in low-stringency BLAST searches, suggesting that this alignments accumulation could be used to highlight coding regions in sequenced DNA. To investigate this possibility, we developed a computer program (AnABlast) that generates profiles of accumulated alignments in query amino acid sequences using a low-stringency BLAST strategy. To validate this approach, all six-frame translations of DNA sequences between every two annotated exons of the fission yeast genome were analysed with AnABlast. AnABlast-generated profiles identified three new copies of known genes, and four new genes supported by experimental evidence. New pseudogenes, ancestral carboxyl- and amino-terminal subtractions, complex gene rearrangements, and ancient fragments of mitDNA and of bacterial origin, were also inferred. Thus, this novel in silico approach provides a powerful tool to uncover new genes, as well as fossil-coding sequences, thus providing insight into the evolutionary history of annotated genomes.


A Commercial Arbuscular Mycorrhizal Inoculum Alleviated the Effects of Acid Water on Lupinus angustifolius Grown in a Sterilized Mining Dump.
Lupinus species have been sporadically reported to be colonized by arbuscular mycorrhizal fungi (AMF). The interactions between AMF and lupine plants could also be non-symbiotic, from positive to negative, as controlled by the stress conditions of the plant. The goal of the study was to reveal the existence of such positive interactions and provide preliminary data for a myco-phytoremediation technology of mining dumps using L. angustifolius as a first crop. The objective was to test the hypothesis that the AMF inoculation of an acidified dump material contaminated with heavy metals would improve the growth of L. angustifolius and decrease oxidative stress. The design consisted of a one-month bivariate pot experiment with plants grown in a mining dump soil inoculated and not inoculated with a commercial AMF inoculum sequestered in expanded clay and watered with acidic and neutral water. There was no AMF root colonization under the experimental conditions, but under neutral and acidic water conditions, the phosphorus concentrations in roots and leaves increased, and the superoxide dismutase and peroxidase activities significantly decreased due to AMF inoculation. The increase in leaf phosphorus concentration was correlated with the decrease in peroxidase activity. The fresh weight of shoots and leaves significantly increased due to the commercial inoculum (under acidic water conditions). At the end of the experiment, the ammonium concentration in the substrate was higher in the inoculated treatments than in the not inoculated ones, and the concentrations of many elements in the dump material decreased compared to the start of the experiment. A comprehensive discussion of the potential mechanisms underlying the effects of the commercial AMF inoculum on the non-host L. angustifolius is completed.


Food intake diet and sperm characteristics in a blue zone: a Loma Linda Study.
The study examined the effect the life-long vegetarian diet on male fertility and focused on vegetarians living in the Loma Linda blue zone, a demographic area known for life longevity. The objective was to compare sperm characteristics of vegetarian with non-vegetarian males. The cross-sectional observational study was based on semen analyses of 474 males from 2009 to 2013. Patients categorized themselves as either life-long lacto-ovo vegetarians (N=26; vegetable diet with dairy and egg products), vegans (N=5; strictly vegetables with no animal products) or non-vegetarians (N=443; no diet restrictions). Sperm quality was assessed using a computer-aided sperm analyzer and strict morphology and chromatin integrity were manually evaluated. Lacto-ovo vegetarians had lower sperm concentration (50.7±7.4M/mL versus non-vegetarians 69.6±3.2M/mL, mean±S.E.M.). Total motility was lower in the lacto-ovo and vegan groups (33.2±3.8% and 51.8±13.4% respectively) versus non-vegetarians (58.2±1.0%). Vegans had lowest hyperactive motility (0.8±0.7% versus lacto-ovo 5.2±1.2 and non-vegetarians 4.8±0.3%). Sperm strict morphologies were similar for the 3 groups. There were no differences in rapid progression and chromatin integrity. The study showed that the vegetables-based food intake decreased sperm quality. In particular, a reduction in sperm quality in male factor patients would be clinically significant and would require review. Furthermore, inadequate sperm hyperactivation in vegans suggested compromised membrane calcium selective channels. However, the study results are cautiously interpreted and more corroborative studies are needed.


Hemoglobin-mediated oxidation of washed minced cod muscle phospholipids: effect of pH and hemoglobin source.
Lipid pro-oxidative properties and deoxygenation/autoxidation patterns of hemoglobins from nonmigratory white-fleshed fish (winter flounder and Atlantic pollock) and migratory dark-fleshed fish (Atlantic mackerel and menhaden) were compared during ice storage at pH 7.2 and 6. A washed cod mince model system and a buffer model system were used for studying lipid changes and hemoglobin changes, respectively. TBARS and painty odor were followed as markers for lipid oxidation. At pH 6, all four hemoglobins were highly and equally active as pro-oxidants. At pH 7.2, pro-oxidation by all hemoglobins except that from pollock was slowed down, and activity ranked as pollock > mackerel > menhaden > flounder. The higher catalytic activities of the hemoglobins at pH 6 than at pH 7.2 corresponded with higher formation of deoxyhemoglobin and methemoglobin. Pollock had the most extensive formation of deoxy- and methemoglobin at both pH values, which could explain its high catalytic activity. The pro-oxidative differences among the other hemoglobins at pH 7.2 did not correlate with deoxygenation and autoxidation reactions. This indicates involvement of other structural differences between the hemoglobins such as differences in the heme-crevice volume. It is suggested that a biological reason for the species differences was their adaptations to different depths/water temperatures.


EFFECTS OF PLANT PROTEIN DIETS ON THE HEALTH OF FARMED AMERICAN ALLIGATORS (ALLIGATOR MISSISSIPPIENSIS).
The objective of this prospective, blinded study was to compare plasma biochemical values and gross and histologic evaluation of kidney and liver from American alligators ( Alligator mississippiensis ) fed extruded diets with protein derived from animal or plant sources. Alligators in two treatment groups were fed an extruded diet with protein derived primarily from plant products for 7 (n = 20) or 10 (n = 20) mo prior to harvest. A control group (n = 20) was fed a commercial diet with protein derived from animal products for the duration of the study. Plasma biochemistry panels were obtained and gross and histologic examination of kidney and liver tissues was conducted for each animal. No differences were found between alligators fed diets with animal or plant protein in terms of either biochemistry profiles or gross or histologic examination of kidney and liver. Plant-based diets, fed for up to 10 mo, do not appear to have any ill effects on the kidney or liver of American alligators.


Posterior medial meniscus root ligament lesions: MRI classification and associated findings.
The purposes of this study were to determine the prevalence of altered MRI appearances of "posterior medial meniscus root ligament (PMMRL)" lesions, introduce a classification of lesion types, and report associated findings. We retrospectively reviewed 419 knee MRI studies to identify the presence of PMMRL lesions. Classification was established on the basis of lesions encountered. The medial compartment was assessed for medial meniscal tears in the meniscus proper, medial meniscal extrusion, insertional PMMRL osseous changes, regional synovitis, osteoarthritis, insufficiency fracture, and cruciate ligament abnormality. PMMRL abnormalities occurred in 28.6% (120/419) of the studies: degeneration, 14.3% (60/419) and tear, 14.3% (60/419). Our classification system included degeneration and tearing. Tearing was categorized as partial or complete with delineation of the point of failure as entheseal, midsubstance, or junction to meniscus. Of all tears, 93.3% (56/60) occurred at the meniscal junction. Univariate analysis revealed significant differences between the knees with and without PMMRL lesions in age, medial meniscal tear, medial meniscal extrusion, insertional PMMRL osseous change, regional synovitis, osteoarthritis, insufficiency fracture (p=0.017), and cruciate ligament degeneration (p<0.001). PMMRL lesions are commonly detected in symptomatic patients. We have introduced an MRI classification system. PMMRL lesions are significantly associated with age, medial meniscal tears, medial meniscal extrusion, insertional PMMRL osseous change, regional synovitis, osteoarthritis, insufficiency fracture, and cruciate ligament degeneration.


Spatial patterns of atrophy, hypometabolism, and amyloid deposition in Alzheimer's disease correspond to dissociable functional brain networks.
Recent neuroimaging studies of Alzheimer's disease (AD) have emphasized topographical similarities between AD-related brain changes and a prominent cortical association network called the default-mode network (DMN). However, the specificity of distinct imaging abnormalities for the DMN compared to other intrinsic connectivity networks (ICNs) of the limbic and heteromodal association cortex has not yet been examined systematically. We assessed regional amyloid load using AV45-PET, neuronal metabolism using FDG-PET, and gray matter volume using structural MRI in 473 participants from the Alzheimer's Disease Neuroimaging Initiative, including preclinical, predementia, and clinically manifest AD stages. Complementary region-of-interest and voxel-based analyses were used to assess disease stage- and modality-specific changes within seven principle ICNs of the human brain as defined by a standardized functional connectivity atlas. Amyloid deposition in AD dementia showed a preference for the DMN, but high effect sizes were also observed for other neocortical ICNs, most notably the frontoparietal-control network. Atrophic changes were most specific for an anterior limbic network, followed by the DMN, whereas other neocortical networks were relatively spared. Hypometabolism appeared to be a mixture of both amyloid- and atrophy-related profiles. Similar patterns of modality-dependent network specificity were also observed in the predementia and, for amyloid deposition, in the preclinical stage. These quantitative data confirm a high vulnerability of the DMN for multimodal imaging abnormalities in AD. However, rather than being selective for the DMN, imaging abnormalities more generally affect higher order cognitive networks and, importantly, the vulnerability profiles of these networks markedly differ for distinct aspects of AD pathology.


Wavelet adaptive backstepping control for a class of nonlinear systems.
This paper proposes a wavelet adaptive backstepping control (WABC) system for a class of second-order nonlinear systems. The WABC comprises a neural backstepping controller and a robust controller. The neural backstepping controller containing a wavelet neural network (WNN) identifier is the principal controller, and the robust controller is designed to achieve L2 tracking performance with desired attenuation level. Since the WNN uses wavelet functions, its learning capability is superior to the conventional neural network for system identification. Moreover, the adaptation laws of the control system are derived in the sense of Lyapunov function and Barbalat's lemma, thus the system can be guaranteed to be asymptotically stable. The proposed WABC is applied to two nonlinear systems, a chaotic system and a wing-rock motion system to illustrate its effectiveness. Simulation results verify that the proposed WABC can achieve favorable tracking performance by incorporating of WNN identification, adaptive backstepping control, and L2 robust control techniques.


Chemopreventive effect of a vitamin D(3) analog, alfacalcidol, on colorectal carcinogenesis in mice with ulcerative colitis.
An increased incidence of colorectal carcinoma is known to occur in patients with ulcerative colitis (UC), which displays a cycle of recurrence-remission in the colorectal mucosa. Repeated oral doses of 3% dextran sulfate sodium subsequent to a single intraperitoneal injection of azoxymethane induced a chronic UC resulting in an increased incidence of high-grade dysplasia and submucosal-invasive adenocarcinomas in the mouse colorectum. The active form of vitamin D(3) is a calcium-regulating hormone that increases serum calcium levels and intestinal calcium absorption. It has been reported that there is an inverse correlation between serum levels of the active metabolite of vitamin D and colorectal carcinoma stage. The features of the colitis induced in this animal model are very similar to the UC in patients in terms of both clinical and histological characteristics. Treatment with a vitamin D(3) analog, alfacalcidol, in mice prevented colitis and carcinogenesis; this is shown by inhibition of the decrease in colorectal length and inhibition of the increased incidence of colorectal dysplasia, with a reduction in the mRNA expression of the DNA-synthesizing enzyme, thymidine kinase, in colorectal tissues.


[The change of the anti-VEGF agent in the treatment of exceptionally active choroidal neovascular membrane in age-related macular degeneration].
Aim of this case report to present a case a male patient with age related macular degeneration succesfully treated by anti-VEGF substances different types. The autor presents a case of 83 year old male with active choroidal neovasculare membrane. In this patient, the best corrected visual acuity (BCVA) of the left eye was 4/10 and the central retina thickness of the left eye was 320 microm. Activity of neovascular membrane was verified by OCT and FAG. Treatment was started with anti-VEGF Macugen. After 2 intravitreal injections of Macugen was state of retina worse. Treatment of the left eye was terminated according to recommendation in Information about drug Macugen. After that was started intravitreal applications of Lucentis. The state of retina was stabilizated after four injections of Lucentis. BCVA of the left eye was 4/20 and the central retina thickness was 130 microm. It happend after one year from beginning of therapy. Dosing of anti-VEGF substances results from clinical trials is suitable for treatment majority of patients with choroidal neovascular membrane. Change of anti-VEGF substance is necessary for stabilisation of state on retina in some cases.


Successful sleeve lobectomy of inflammatory myofibroblastic tumor in a 4-year-old child.
Primary pulmonary tumors in small children have remained a challenge for pediatric surgeons. Pneumonectomy and radical lobectomy are limitedly indicated due to surgical difficulties and sequelae. Here, we present our experience with a 4-year-old patient who suffered from an inflammatory myofibroblastic tumor. A left lower sleeve lobectomy was performed, and the patient recovered significantly after surgery. At the last follow-up, the child was growing well without any sequel, which supports our hypothesis that in small children, sleeve resection is the preferred treatment for tumors on the main stem bronchus and presents an alternative to an otherwise unavoidable pneumonectomy.


Neural mechanisms of vocal imitation: The role of sleep replay in shaping mirror neurons.
Learning by imitation involves not only perceiving another individual's action to copy it, but also the formation of a memory trace in order to gradually establish a correspondence between the sensory and motor codes, which represent this action through sensorimotor experience. Memory and sensorimotor processes are closely intertwined. Mirror neurons, which fire both when the same action is performed or perceived, have received considerable attention in the context of imitation. An influential view of memory processes considers that the consolidation of newly acquired information or skills involves an active offline reprocessing of memories during sleep within the neuronal networks that were initially used for encoding. Here, we review the recent advances in the field of mirror neurons and offline processes in the songbird. We further propose a theoretical framework that could establish the neurobiological foundations of sensorimotor learning by imitation. We propose that the reactivation of neuronal assemblies during offline periods contributes to the integration of sensory feedback information and the establishment of sensorimotor mirroring activity at the neuronal level.


High fat diet and high polyphenols beverages effects in enzymatic and non-enzymatic antioxidant activity.
High fat diets have been implicated in the generation of reactive oxygen species (ROS). Polyphenols from grapes may reduce ROS and restore oxidative balance. The aim of this study is to investigate the antioxidant properties of high polyphenols beverages associated with a high fat diet in enzymatic and non-enzymatic antioxidant activity. Fifty female rats were divided into five groups: a) control group (CG) - control diet (4% fat); b) high fat diet group (HFD) - high fat diet (20% fat); c) grape juice group (GJ) - grape juice (15 ml/day) + high fat diet; d) red wine group (RW) - red wine (10 ml/day) + high fat diet; and e) resveratrol solution group (RS) - resveratrol solution (15 ml/day) + high fat diet. Eight weeks later, the superoxide dismutase, catalase and glutathione peroxidase activities were measured. Superoxide dismutase activity was assayed by measuring the inhibition of adrenaline auto-oxidation, catalase by the decrease rate in hydrogen peroxide and glutathione peroxidase by monitoring the oxidation of nicotinamide adenine dinucleotide phosphate. Non-enzymatic antioxidant activity was assessed by oxygen radical absorbance capacity and DDPH (free radical sequestration 2,2-diphenyl-1-picryhydracil) method in the animal's plasma. GC and GJ presented the lowest glutathione peroxidase activity, pointing to a possible protective effect of grape juice against high levels of ROS (p < 0.05). RW increased catalase activity when compared to the RS (p <0 .05). Superoxide dismutase activity and non-enzymatic antioxidant plasma activity were similar in all groups. Grape juice showed to be the most effective in minimizing the deleterious effects of a high fat diet. Resveratrol did not present any benefit and red wine possibly shows a harmful effect due to ethanol content.


Molecular Cloning, Expression Profiling, and Marker Validation of the Chicken Myoz3 Gene.
Myozenin3 (Myoz3) has been reported to bind multiple Z-disc proteins and hence play a key role in signal transduction and muscle fiber type differentiation. The purpose of current study is to better understand the basic characteristics of Myoz3. Firstly, we cloned the ORF (open reading frame) of the Myoz3 gene. AA (amino acid) sequence analysis revealed that the Myoz3 gene encodes a 26 kDa protein which have 97% identities with that of turkey. Expression profiling showed that Myoz3 mRNA is mainly expressed in leg muscle and breast muscle. Furthermore, we investigated Myoz3 gene polymorphisms in two broiler breeds, the Yellow Bantam (YB) and the Avian. Five SNPs (single nucleotide polymorphisms) were identified in the YB breed and 3 were identified in the Avian breed. Genotypes and haplotype were constructed and their associations with carcass traits were analyzed. In the YB breed, c.516 C>T had a strong effect on both shank bone length and the [Formula: see text] value of breast muscle, and the H1H3 diplotype had the highest FC compared to other diplotypes. The markers identified in this study may serve as useful targets for the marker-assisted selection (MAS) of growth and meat quality traits in chickens.


Decline in serum albumin concentration is a predictor of serious events in nonalcoholic fatty liver disease.
Nonalcoholic fatty liver disease (NAFLD) is associated with metabolic syndrome, which includes diabetes mellitus and hyperlipidemia. A fraction of NAFLD patients develop nonalcoholic steatohepatitis, leading to cirrhosis associated with various serious complications, including hepatocellular carcinoma, gastroesophageal varices, cardiovascular events, and other organ malignancy. Although the incidence of chronic viral hepatitis with associated complications has gradually decreased as highly effective antiviral therapies have been established, the number of patients with steatohepatitis has been increasing.This retrospective study examined data of 229 patients from 22 hospitals in our region. We examined 155 cases of chronological data and assessed the development of liver fibrosis and evaluated hepatic reserve-related markers such as platelet count, FIB-4 index, prothrombin time, and serum albumin concentration. We analyzed the relationship of these chronological changes and the incidence of NAFLD related serious complications.Data related to liver fibrosis progression, albumin, and prothrombin time were significantly associated with the occurrence of serious complications associated with cirrhosis. We compared 22 event and 133 nonevent cases of chronological changes in the data per year and found that serum albumin concentration was significantly lower in the group that developed serious complications (event cases: -0.21 g/dL/year, nonevent cases: -0.04 g/dL/year (P < .001)). This albumin decline was only the associated factor with the event incidence by multivariate analysis (P < .01).Annual decline in serum albumin concentration in patients with NAFLD is associated with serious events from the outcome of multicenter retrospective study. This highlights its potential utility as a surrogate marker to assess the efficacy of prediction of NAFLD related serious events.


Analyses of microbial activity in biomass-recycle reactors using denaturing gradient gel electrophoresis of 16S rDNA and 16S rRNA PCR products.
The relationship between mixed microbial community structure and physiology when grown under substrate-limited conditions was investigated using continuous-flow bioreactors with 100% biomass recycle. Community structure was analyzed by denaturing gradient gel electrophoresis (DGGE) of the PCR and RT-PCR amplified V3 region of 16S rDNA and 16S rRNA templates, respectively. Comparisons were made of communities exposed to different types of transient conditions (e.g., long- and short-term starvation, increasing nutrients). With progressively more stringent substrate limitation over time, the specific content of community RNA declined by more than 10-fold and closely followed the decline in specific growth rate. In contrast, the DNA content was variable (up to 3-fold differences) and did not follow the same trend. Cluster analysis of the presence or absence of individual bands indicated that the fingerprints generated by the two templates were different, and community response was first observed in the rRNA fraction. However, both the rDNA and rRNA fingerprints provided a picture of temporal population dynamics. Dice similarity coefficients gave a quantitative measure of the differences and changes between the communities. In comparison, standard cultivation techniques yielded only a quarter of the phylotypes detected by DGGE, but included the most dominant population based on rRNA. Nucleotide-sequence analyses of the almost complete 16S rRNA genes of these isolates place them in the same group of organisms that is typically cultivated from environmental samples: alpha, beta, and gamma Proteobacteria and the high GC and the low GC Gram-positive divisions.


On the Assessment of Surface Quality and Productivity Aspects in Precision Hard Turning of AISI 4340 Steel Alloy: Relative Performance of Wiper vs. Conventional Inserts.
This article reports an experimental assessment of surface quality generated in the precision turning of AISI 4340 steel alloy using conventional round and wiper nose inserts for different cutting conditions. A three-factor (each at 4 levels) full factorial design of experiment was followed for feed rate, cutting speed, and depth of cut, with resulting machined surface quality characterized by resulting average roughness (Ra). The results show that, for the provided range of cutting conditions, lower surface roughness values were obtained using wiper inserts compared with conventional inserts, indicating a superior performance. When including the type of insert as a qualitative factor, ANOVA revealed that the type of insert was most important in determining surface roughness and material removal rate, with feed rate as the second most significant, followed by the interaction of feed rate and type of insert. It was found that using wiper inserts allowed simultaneous increases in feed rate, cutting speed, and depth of cut, while providing better surface quality of lower Ra, compared to the global minimum value that could be achieved using the conventional insert. These findings show that wiper inserts produce better surface quality and a material removal rate up to ten times higher than that obtained with conventional inserts. This clearly indicates the tremendous advantages of high surface quality and productivity that wiper inserts can offer when compared with the conventional round nose type in precision hard turning of AISI 4340 alloy steel.


A temperature and precipitation record of the past 16,000 years in southern chile.
Regression equations relating pollen taxa from surface samples to temperature and precipitation are applied to a radiocarbon-dated pollen sequence in a lake core from Alerce. The resulting curves are a measure of the fluctuations of these climatic variables and show similarities to other late Quaternary records from the Southern Hemisphere.


Need for simulation in laparoscopic colorectal surgery training.
The dissemination of laparoscopic colorectal surgery (LCS) has been slow despite increasing evidence for the clinical benefits, with a prolonged learning curve being one of the main restrictions for a prompt uptake. Performing advanced laparoscopic procedures requires dedicated surgical skills and new simulation methods designed precisely for LCS have been established: These include virtual reality simulators, box trainers, animal and human tissue and synthetic materials. Studies have even demonstrated an improvement in trainees' laparoscopic skills in the actual operating room and a staged approach to surgical simulation with a combination of various training methods should be mandatory in every colorectal training program. The learning curve for LCS could be reduced through practice and skills development in a riskfree setting.


[The influence of immunosuppression on vancomycin concentration in the rat allogenic lung transplant].
To assess the influence of conventional immunosuppression on vancomycin concentration in the rat allogenic lung transplant basing on acute and hyperacute rejection models. Left lung allotransplantations were performed from Brown Norway donors to Fisher F 344 recipients in the acute rejection model (animals were sacrificed 5 days after transplantation), and from Brown Norway donors to Wistar recipients in the hyperacute rejection model (animals were sacrificed 2 days after transplantation). Immunosuppression (cyclosporin A 5 mg/kg b.w., aziathioprine 4 mg/kg b.w., methylprednisolone 4 mg/kg b.w.) was administered daily. Control rats received no immunosuppression and were sacrificed on day 2 or 5, respectively. Rejection grading was done on the basis of arterial pO2 and histology of the lung graft sample obtained at autopsy. A single 30 mg/kg b.w. dose ofvancomycin was injected intraperitoneally on day 2 or 5 depending on the model. Samples of blood and grafted lung were collected 30 min, 1 h, 2 h, 4 h, and 6 h from injection. Arterial pO2 levels were significantly higher in the group with acute rejection and immunosuppression as compared with the control groups. Histology revealed attenuated rejection in the immunosuppression groups. Vancomycin concentration in the transplanted lung and the lung graft to plasma vancomycin concentration ratio did not differ in the immunosuppression and control groups. Immunosuppression has no influence on vancomycin concentration in the transplanted lung.


Modeling Shift-Variant X-Ray Focal Spot Blur for High-Resolution Flat-Panel Cone-Beam CT.
Flat-panel cone-beam CT (CBCT) has been applied clinically in a number of high-resolution applications. Increasing geometric magnification can potentially improve resolution, but also increases blur due to an extended x-ray focal-spot. We present a shift-variant focal-spot blur model and incorporate it into a model-based iterative-reconstruction algorithm. We apply this algorithm to simulation and CBCT test-bench data. In a trabecular bone simulation study, we find traditional reconstruction approaches without a blur model exhibit shift-variant resolution properties that depend greatly on the acquisition protocol (e.g. short vs. full scans) and the anode angles of the rays used to reconstruct a particular region. For physical CBCT experiments focal spot blur was characterized and a spatial resolution phantom was scanned and reconstructed. In both experiments image quality using the shift-variant model was significantly improved over approaches that modeled no blur or only a shift-invariant blur, suggesting a potential means to overcome traditional CBCT spatial resolution and system design limitations.


Effect of the Size and Shape on the Electrocatalytic Activity of Co3O4 Nanoparticles in the Oxygen Evolution Reaction.
Co3O4 nanoparticles were size- and shape-selectively synthesized in a solvothermal approach by the thermal decomposition of either cobalt(II) acetylacetonate [Co(acac)2] or cobalt(II) nitrate [Co(NO3)2] in different solvents followed by calcination under air. Spherical, cubic, octahedral, and platelike nanoparticles with narrow size distributions and sizes ranging from 8 to 90 nm were formed in high yield. The nanoparticles were characterized by X-ray diffraction, scanning and transmission electron microscopy, energy-dispersive X-ray analysis, and X-ray photoelectron spectroscopy. The distinctive influence of the size and shape of the nanoparticles on the electrocatalytic activity in the oxygen evolution reaction is also demonstrated.


[Forces at the implant-bone interface of condylar knee prosthesis--with special reference to the retention of the posterior cruciate ligament].
Breakdown of the implant-bone interface is the most important factor limiting the effective life of total joint replacement. A particular example is the tibial component of the knee prosthesis which interfaces with trabecular bone. It is postulated that one of the major reasons for interface breakdown is excessive shear, compressive and moment stresses at the implant-bone interface. These stresses can be greatly influenced by the design of the prostheses, namely surface contour of the tibial component and the retention of the posterior cruciate ligament. Forces across the tibial component-bone interface was measured for prostheses both retaining and sacrificing the posterior cruciate ligament and for partially conforming and non-conforming, flat geometries. The results are as follows: Shear force, normal force and moment force across the interface were significantly greater for the prostheses sacrificing the posterior cruciate ligament than for those retaining it. Moment force was markedly greater for the tibial component with anterior curvature than for flat tibial component. It was suggested from this experiment that the knee prosthesis should retain the posterior cruciate ligament and the surface of the tibial component should probably be flatter for reducing the forces at the implant-bone interface.


[Presentation of a clinical hemoviscosimeter].
A clinical hemoviscosiSImeter is presented. It is a Couette type viscometer which is automatic and driven by a microcomputer. It was designed and realized in the laboratory. The measurement of blood viscosity requires 1 ml of anticoagulated blood adjusted at 45% of hematocrit. Experiments are performed at a constant temperature (37 degrees C) according to a well-established protocol: gentle mixing of the blood sample followed by a 3 minute rest just before each of the following measurement modes begins. Under steady state flow conditions, 12 viscosity measurements are realized in a range of shear stress rate step of 1 s-1 is applied. The elastothixotropic response of the blood sample is then obtained. Such a hemorheological test, called "hemorheogram", requires 15 minutes. Given to the clinician, the hemorheogram results include the absolute blood viscosities at the shear stress rates of 0.1 s-1, 1 s-1, 10 s-1, 70 s-1, the thixotropy index and the plasma viscosity. Finally, results to be kept in the laboratory files consist of the whole data issued from the patient hemorheogram including the relative blood viscosities (blood viscosity/plasma viscosity) at the same shear strers rate values.


A flexible mathematical model platform for studying branching networks: experimentally validated using the model actinomycete, Streptomyces coelicolor.
Branching networks are ubiquitous in nature and their growth often responds to environmental cues dynamically. Using the antibiotic-producing soil bacterium Streptomyces as a model we have developed a flexible mathematical model platform for the study of branched biological networks. Streptomyces form large aggregates in liquid culture that can impair industrial antibiotic fermentations. Understanding the features of these could aid improvement of such processes. The model requires relatively few experimental values for parameterisation, yet delivers realistic simulations of Streptomyces pellet and is able to predict features, such as the density of hyphae, the number of growing tips and the location of antibiotic production within a pellet in response to pellet size and external nutrient supply. The model is scalable and will find utility in a range of branched biological networks such as angiogenesis, plant root growth and fungal hyphal networks.


Genetic variation in nitric oxide synthase 2A (NOS2A) and risk for multiple sclerosis.
Multiple sclerosis (MS) is a chronic inflammatory disorder of the central nervous system with a strong genetic component. Variation in the major histocompatibility complex on chromosome 6p21, specifically the HLA-DRB1*15 haplotype, is the strongest genetic factor for MS, yet it is estimated to account for only a portion of risk for the disease. Previous evidence has implicated the nitric oxide synthase gene (NOS2A) encoding inducible NOS on chromosome 17q11 as a potential MS susceptibility gene. To determine whether variation in the NOS2A gene contributes to MS risk, we investigated a total of 50 polymorphisms within or flanking the locus for evidence of association using a comprehensive analytical strategy. A total of 6265 members from 1858 well-characterized MS families were utilized. No evidence for overtransmission of any individual single-nucleotide polymorphism allele or haplotype to the MS-affected individuals was observed. Furthermore, different transmission rates were not observed in either DRB1*15-positive or DRB1*15-negative family subgroups, or when extreme clinical outcomes characterizing disease progression were examined. The very largest study of NOS2A variation in MS, to date, excludes even a modest role for this locus in susceptibility.


[Volume measurement of the horizontal extraocular muscles using magnetic resonance imaging].
The volume of the horizontal extraocular muscles of 11 normal adults and three patients with ophthalmoplegia was measured using magnetic resonance imaging (MRI). The MRI examinations were carried out with a Signa Advantage, 1.5 tesla superconductive magnetic system manufactured by General Electric. This method employs the spin echo technique with a 3.0 mm gapless slice, a 350 ms. repetition time, and a 17.0 ms. echo time. The MRI films were projected and magnified on Kent paper using an overhead projector. Then the shapes of the horizontal extraocular muscles were traced. The volume of the muscles was measured as the total weight of Kent papers which were cut out from muscle shapes in all the slices. The average volume of the normal medial and lateral rectus muscles was 690 +/- 87 mm3 and 734 +/- 77 mm3, respectively. Two cases with peripheral nerve palsy showed typical atrophy of the paretic muscles. A case with orbital myositis showed typical hypertrophy of the inflamed muscles. This measurement may prove useful in the analysis and evaluation of extraocular muscles, especially in ophthalmoplegia.


The ethics of nurse migration: an evolution of community change.
International nursing migration, or the movement of people across international boundaries, has enormous implications for the discipline of nursing. This column focuses on ethical assumptions and possible implications for community change as individual or group. Discussion follows with the illumination of community change paradoxes, implications for policy development, nursing education, and nursing practice from the nursing theoretical perspective of the humanbecoming school of thought.


Adult tuberculous meningitis in Qatar: a descriptive retrospective study from its referral center.
Tuberculous meningitis is the severest and commonest form of central nervous system tuberculosis causing high mortality and morbidity. We aim to study the clinical, biochemical, and radiological characteristics of tuberculous meningitis in adult patients in Qatar and to calculate the incidence. A descriptive retrospective 7-year study was conducted at the Hamad General Hospital (the only tertiary referral center and sole health care provider for tuberculosis in Qatar) between the 1st of January 2006 and the 31st of December 2012, to describe the clinical presentation, diagnosis, treatment, outcome, and the incidence of adult tuberculous meningitis in Qatar. Tuberculous meningitis was diagnosed in 80 patients (65 male, 15 female), with a mean age of 30.3 ± 8.9. The majority of patients (76.3%) were from the Indian subcontinent. The commonest clinical features were fever (79%), headache (71%), and meningism (56%). Cerebrospinal fluid tuberculosis culture was positive in 44% of patients. Almost 39% of patients were in Stage I, 46% in Stage II and 15% in Stage III of the disease. The commonest neuroimaging features were leptomeningeal enhancement (34%) and hydrocephalus (33%). Cranial nerve palsy, limb weakness, and an elevated C-reactive protein were associated with a poorer outcome. Sixty eight percent had complete recovery, 10% had residual neurological sequelae without disability, 17% had disability, and 5% died. The average incidence of tuberculous meningitis over 7 years was 0.9 per 100,000 adult population. The characteristics of tuberculous meningitis were described in our population. Our data indicate that the incidence of tuberculosis meningitis in Qatar has increased. Tuberculous meningitis in Qatar is mainly an imported disease.


Enhancement of the click chemistry for the inverse Diels Alder technology by functionalization of amide-based monomers.
In the near future personalized medicine with nucleic acids will play a key role in molecular diagnostics and therapy, which require new properties of the nucleic acids, like stability against enzymatic degradation. Here we demonstrate that the replacement of nucleobases with PNA by functional molecules harbouring either a dienophile or a diene reactivity is feasible and confers all new options for functionalization. These newly developed derivatives allow independent multi-ligations of multi-faceted components by use of the inverse Diels Alder technology. The high chemical stability and the ease of synthesis qualify these polyamide building blocks as favourites for intracellular delivery and targeting applications. This allows local drug concentrations sufficient for imaging and therapy and simultaneously a reduction of the application doses. It is important to point out that this technology is not restricted to ligation of medicament material; it is also a candidate to develop new and highly efficient active compounds for a "sustainable pharmacy".


Interaction between stress induced by competition, predation, and an insecticide on the response of aquatic invertebrates.
The present study investigated the effects of species interactions like competition and (intraguild) predation on the sensitivity of aquatic organisms to the insecticide chlorpyrifos. In the first experiment, combined effects of chlorpyrifos and different levels of intraspecific and interspecific interaction were assessed on Gammarus pulex survival using Asellus aquaticus as an interacting species. Intraspecific and interspecific interactions increased the time to extinction of G. pulex up to a factor of 2, most likely because of the cannibalistic nature of G. pulex and its intraguild predation on A. aquaticus under stress conditions. In the second experiment, combined effects of chlorpyrifos and intraspecific and interspecific interaction were assessed on Daphnia pulex abundance using Brachionus calyciflorus as a competing species and Chaoborus sp. larvae as a predator. Intraspecific and interspecific interactions significantly affected the D. pulex population structure, but they did not influence the total population size. Predation decimated D. pulex abundance; however, interacting effects of predation and chlorpyrifos exposure were less noticeable at high exposure concentrations because of the reduced predatory efficiency of Chaoborus sp. larvae. The present study shows that species interactions do not always increase the vulnerability of aquatic populations to chemical stress and that some interactions (e.g., cannibalism and intraguild predation) or reduced predator grazing pressure can alleviate competition and predation stress on population-level insecticide effects under food-limiting conditions. Environ Toxicol Chem 2017;36:2485-2492. © 2017 SETAC.


Preclinical development and characterisation of 99mTc-NM-01 for SPECT/CT imaging of human PD-L1.
The level of expression of programmed cell death-1 (PD-1)/programmed death ligand-1 (PD-L1) is a predictive biomarker for cancer immunotherapy, however, its detection remains challenging due to tumour heterogeneity and the influence from the binding of therapeutic agents. We recently developed [99mTc]-NM-01 as a companion diagnostic imaging agent for non-invasive molecular imaging of PD-L1 by single-photon emission computed tomography (SPECT). The aim of the study was to evaluate the [99mTc] radiolabelling of GMP graded NM-01 and its pharmacology, pharmacokinetics and toxicology. NM-01 bound specifically to human PD-L1 (Kd=0.8 nM) and did not interfere with the binding of the anti-PD-L1 antibody atezolizumab. NM-01 can bind various PD-L1-positive cancer cell lines and only interact with PD-L1 expressed on the cell surface. In SPECT/CT imaging, high [99mTc]-NM-01 accumulation was observed in the HCC827 mouse xenografted tumour model (30-min: 1.50 ± 0.27 %ID/g; 90-min: 1.23 ± 0.18 %ID/g), demonstrated a predominantly renal elimination (high uptake in bladder and kidney), while activity in the blood pool and other major organs remained low. The tumour-to-muscle and tumour-to-blood ratios were comparable with/without atezolizumab (P<0.04) but were significantly lowered when co-injected with excess NM-01 (P=0.04 and P=0.01, respectively.) The blood clearance of [99mTc]-NM-01 is bi-phasic; consisting of an initial fast washout phase with half-life of 2.1 min and a slower clearance phase with half-life of 25.4 min. In an intravenous extended single-dose toxicity study, no treatment-related changes were observed and the maximum tolerated dose of [99mTc]-NM-01 was 2.58 mg/kg. [99mTc]-NM-01 has suitable properties as a potential candidate for SPECT/CT imaging of PD-L1 assessment in cancer patients.


Holothurian glycosaminoglycan inhibits metastasis and thrombosis via targeting of nuclear factor-κB/tissue factor/Factor Xa pathway in melanoma B16F10 cells.
Holothurian glycosaminoglycan (hGAG) is a high-molecular-weight form of fucosylated chondroitin sulfate and has an antithrombotic effect. Our previous studies demonstrated that hGAG efficiently inhibited tumor cell metastasis. The interplays between thrombosis and tumor progression may have a major impact on hematogenous metastasis. In this study, we demonstrated that the mouse melanoma B16F10 cells treated with hGAG displayed a significant reduction of metastasis and coagulation capacity in vitro and in vivo. Mechanistic studies revealed that hGAG treatment in B16F10 cells remarkably inhibited the formation of fibrin through attenuating the generation of activated Factor Xa (FXa), without affecting the expression of urokinase (uPA) and plasminogen activator inhibitor 1 (PAI-1) that involved in fibrinolysis. Moreover, hGAG treatment downregulated the transcription and protein expression of tissue factor (TF). Promoter deletions, site mutations and functional studies identified that the nuclear transcription factor NF-κB binding region is responsible for hGAG-induced inhibition of TF expression. While the hGAG treatment of B16F10 cells was unable to inhibit NF-κB expression and phosphorylation, hGAG significantly prevented nuclear translocation of NF-κB from the cytosol, a potential mechanism underlying the transcriptional suppression of TF. Moreover, hGAG markedly suppressed the activation of p38MAPK and ERK1/2 signaling pathways, the central regulators for the expression of metastasis-related matrix metalloproteinases (MMPs). Consequently, hGAG exerts a dual function in the inhibition of metastasis and coagulation activity in mouse melanoma B16F10 cells. Our studies suggest hGAG to be a promising therapeutic agent for metastatic cancer treatment.


Application of the TRS 483 code of practice for reference and relative dosimetry in tomotherapy.
To apply the recent code of practice from the IAEA/AAPM, TRS 483, to helical tomotherapy (HT) for reference and relative dosimetry obtaining correction factors for the Exradin A1SL ionization chamber. The beam quality correction factor for the A1SL chamber was obtained through three different approaches following TRS 483 concepts and compared with published values. The determination of the reference absolute dose for the machine-specific reference (msr) field was complemented with relative dosimetry through the determination of output factors of small fields using different detectors. The response of A1SL was compared with correction-free film results and corrected output factors of other detectors. A weighted mean beam quality correction factor of 0.9945± 0.0073 was obtained for the A1SL chamber which is in agreement with values reported in the literature. Output factors obtained with different detectors were in agreement, given the uncertainty level. Considering the film output factors as free of corrections, the average value for A1SL output factors corrections was 1.000 ± 0.007. The beam quality correction factors for the A1SL chamber obtained through the three different pathways recommended by TRS 483 agreed with each other and also with published values. The measurements from the A1SL chamber normalized to the msr field in HT can be taken as output factors for small clinical field sizes without further corrections.


Multiresponse optimization of an extraction procedure of carnosol and rosmarinic and carnosic acids from rosemary.
A green solvent-based optimization for rosmarinic acid (RA), carnosol (COH), and carnosic acid (CA) extraction, the three main antioxidants from rosemary, was performed. The conventional solid-liquid extraction was optimized using a central composite design (CCD) followed by the desirability approach. In the CCD analysis the quantitative effects of extraction time (4.8-55.2min), liquid-to-solid ratio (4.6-21.4mLg(-1)), and ethanol content (44.8-95.2% v/v) were determined for the extracted amount of antioxidants, their concentrations in the extract, and the extraction yield. Samples were analyzed by HPLC and the antioxidants were identified by comparison with pure standard retention times and UV spectra. The desirability function that simultaneously maximizes the antioxidants extraction and their concentrations in the final product was validated. The extraction using a hydroalcoholic solution 70% v/v, at low liquid-to-solid ratio (5mLg(-1)), and after 55-min yielded an antioxidant recovery rate of 89.8%, and a final product 4.75 times richer in the main antioxidants than the raw material.


Dexamethasone alleviates meconium-induced airway hyperresponsiveness and lung inflammation in rabbits.
The effects of dexamethasone on in vitro airway reactivity associated with lung inflammation were investigated in rabbits with meconium aspiration. Oxygen-ventilated adult rabbits received an intratracheal bolus of 4 ml/kg body weight of saline (Sal, n = 4) or human meconium (25 mg/ml). Thirty minutes later, meconium-instilled animals intravenously received 0.5 mg/kg of dexamethasone (Dexa, n = 6), or were left without treatment (Meco, n = 5). The animals were ventilated for a further 5 hr and then sacrificed. The left lungs were lavaged with saline, and the white blood cell (WBC) count was estimated. Tracheal and right-lung tissue strips were placed into organ chambers with Krebs-Henseleit solution. Cumulative doses of histamine (10(-8)-10(-3) mol/l) and acetylcholine (10(-8)-10(-3) mol/l) were added to the chambers, and recordings of contractions were made after a 30-min loading phase with a tension of 4 grams, and another 30-min adaptation phase with a tension of 2 g. Tracheal smooth muscle in vitro reactivity to histamine was higher in the Meco than in the Sal group, and dexamethasone decreased the reactivity compared to the Meco group (P < 0.05). Lung tissue in vitro reactivity to histamine was slightly higher in the Meco than in the Sal group (P > 0.05), and dexamethasone decreased the reactivity compared to both the Meco and Sal groups (P < 0.05). No between-group differences were observed in tracheal or lung in vitro reactivity to acetylcholine (P > 0.05). In the Meco group, blood WBC (P > 0.05) and neutrophil (P < 0.05) counts were lower than in the Sal and Dexa groups. Lung neutrophils and eosinophils were higher in both the Meco and Dexa groups than in the Sal group (P < 0.01). Dexamethasone decreased neutrophils (P < 0.05) compared to the Meco group. Meconium-induced airway hyperreactivity to histamine and lung inflammation were alleviated by dexamethasone.


Proposition of an aging indicator from general health examination in France.
We present an indicator of aging based on eight quantitative variables, measured during the periodical health examination offered by French social security to the general population in France. The used sample derived from the population examined each year. We selected a reference group of 24,510 adults older than 25 years (non-smokers, drinking less than 44 g alcohol per day, with body mass index less than 35, and not using medication). A multiple regression analysis by gender was used to select significant variables to predict age from all quantitative variables measured during the health examination. The resulting biological age could be calculated from the measured variables and the aging indicator was defined as the difference between the calculated age and the chronological age. By definition, this indicator is independent of age in the reference population and gives an indication of the difference between an individual's status in the process of aging and the mean status of people of the same chronological age. Associations between this indicator and life style habits such as tobacco and alcohol use, and occupational status were observed. A geographical analysis also exhibited significant variation over France. Such an indicator can be used at the individual level as a pedagogical tool to explain the observed individual health status. It is also of interest for the epidemiological studies, where it could contribute to a better understanding of the aging process and associated factors.


Dual NEP/ECE inhibition improves endothelial function in mesenteric resistance arteries of 32-week-old SHR.
Endothelin 1 (ET-1), a potent vasoconstrictor, pro-mitogenic and pro-inflammatory peptide, may promote development of endothelial dysfunction and arterial remodeling. ET-1 can be formed through cleavage of big-ET-1 by endothelin-converting enzyme (ECE) or neutral endopeptidase (NEP). We investigated whether chronic treatment with the novel dual NEP/ECE inhibitor SOL1 improves functional and structural properties of resistance-sized arteries of 32-week-old male spontaneously hypertensive rats (SHR). SHR received a chronic 4-week treatment with SOL1, losartan or hydralazine. We then compared effects of inhibition of NO synthase (NOS) (100 μM l-NAME), blockade of ETA- and ETB-receptors (10 μM bosentan) and stimulation of the endothelium with 0.001-10 μM acetylcholine (ACh) in isolated third-order mesenteric resistance arteries. Losartan and hydralazine significantly lowered blood pressure. Losartan decreased the media-to-lumen ratio of resistance arteries. l-NAME (1) increased arterial contractile responses to K+ (5.9-40 mM) in the losartan, SOL1 and vehicle group and (2) increased the sensitivity to phenylephrine (PHE; 0.16-20 μM) in the SOL1 group but not in the losartan, hydralazine and vehicle group. Relaxing responses to ACh in the absence or presence of l-NAME during contractions induced by either 10 μM PHE or 40 mM K+ were not altered by any in vivo treatment. Acute treatment with bosentan did, however, significantly improve maximal relaxing responses involving endothelium-derived nitric oxide and -hyperpolarizing factors in the SOL1 group but not in the losartan, hydralazine or vehicle group. Thus, chronic inhibition of NEP/ECE improved basal endothelial function but did not alter blood pressure, resistance artery structure and stimulated endothelium-dependent relaxing responses in 32-week-old SHR.


Down-regulation of cytokine production and interleukin-2 receptor expression by pooled human IgG.
The influence of pooled human IgG preparations for intravenous use (i.v.Ig) on in vitro-induced cytokine production was studied at the single-cell level using cytokine-specific monoclonal antibodies (mAb) and indirect immunofluorescent technique. Cultured mononuclear cells from peripheral blood from healthy adult donors were polyclonally stimulated for 96 hr by either direct ligation of T-cell receptors using immobilized anti-CD3 mAb or by a combination of a protein kinase C activator [phorbol 12-myristate 13-acetate (PMA)] and a calcium ionophore (ionomycin) in the absence or presence of i.v.Ig. A marked inhibition of proliferation and blast transformation was noted in all i.v.Ig exposed cultures, despite good cell survival. The production of the T-cell lymphokines interleukin-2 (IL-2), IL-10,interferon-gamma (IFN-gamma) and tumour necrosis factor-beta (TNF-beta) was significantly down-regulated during the whole studied period in the i.v.Ig containing anti-CD3 stimulated cultures. The synthesis of the monokine IL-8 was not suppressed and that of TNF-alpha, which was made by both lymphocytes and monocytes, was only moderately inhibited. Somewhat different and more transient effects were observed in the i.v.Ig-exposed PMA/ionomycin-activated cultures. The production of IL-2, IL-3, IL-4, IL-5, IL-10, TNF-beta and granulocyte-macrophage colony-stimulating factor (GM-CSF) was down-regulated during the initial phase of the cultures up to 48 hr, but not at 48-96 hr. The synthesis of IFN-gamma and TNF-alpha was unaffected of the influence of i.v.Ig during the entire culture period. The expression of IL-2 receptors (IL-2R) was significantly suppressed in the i.v.Ig-treated anti-CD3-activated cells, but not in the PMA/ionomycin-stimulated cultures. Taken together our results indicate that pooled IgG may mediate immunomodulation by direct effects on cytokine production and on T-cell proliferation.


Sleep and next-day negative affect and suicidal ideation in borderline personality disorder.
Sleep disturbance is associated with elevated suicidal ideation and negative affect. To date, however, no study has investigated the temporal relationship between sleep and suicidality among those diagnosed with borderline personality disorder (BPD). This preregistered (https://osf.io/4vugk) study tested whether nightly sleep (self-reported sleep duration, sleep onset latency, and subjective sleep quality) represents a (within-person) short-term risk factor for affective dysregulation and increases in suicide risk from day-to-day, as well as whether between-person differences in sleep, negative affect, and suicidality were associated. We used a 21-day ecological momentary assessment protocol in a sample of 153 people diagnosed with BPD, 105 of which had a history of serious suicide attempts, and 52 healthy controls (N = 4076 days). We found a within-person association between worse subjective sleep quality and greater next-day negative affect. At the between-person level, we found positive relationships between sleep latency and suicidal ideation, and a negative association between subjective sleep quality and negative affect. BPD severity did not significantly moderate the strength of any within-person associations, although BPD was positively associated with average levels of suicidal ideation, sleep latency, and negative affect, and negatively related to subjective sleep quality. These findings suggest that the association of sleep with suicidal ideation and BPD exists largely at the between-persons rather than the within-person level. Disturbed sleep, therefore, seems to largely coincide, rather than specifically contribute to, the exacerbation of suicidal crises in BPD. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


RNF168-mediated localization of BARD1 recruits the BRCA1-PALB2 complex to DNA damage.
DNA damage prompts a diverse range of alterations to the chromatin landscape. The RNF168 E3 ubiquitin ligase catalyzes the mono-ubiquitination of histone H2A at lysine (K)13/15 (mUb-H2A), forming a binding module for DNA repair proteins. BRCA1 promotes homologous recombination (HR), in part, through its interaction with PALB2, and the formation of a larger BRCA1-PALB2-BRCA2-RAD51 (BRCA1-P) complex. The mechanism by which BRCA1-P is recruited to chromatin surrounding DNA breaks is unclear. In this study, we reveal that an RNF168-governed signaling pathway is responsible for localizing the BRCA1-P complex to DNA damage. Using mice harboring a Brca1CC (coiled coil) mutation that blocks the Brca1-Palb2 interaction, we uncovered an epistatic relationship between Rnf168- and Brca1CC alleles, which disrupted development, and reduced the efficiency of Palb2-Rad51 localization. Mechanistically, we show that RNF168-generated mUb-H2A recruits BARD1 through a BRCT domain ubiquitin-dependent recruitment motif (BUDR). Subsequently, BARD1-BRCA1 accumulate PALB2-RAD51 at DNA breaks via the CC domain-mediated BRCA1-PALB2 interaction. Together, these findings establish a series of molecular interactions that connect the DNA damage signaling and HR repair machinery.


Determinants of the heart rate variability in type 1 diabetes mellitus.
Evaluation of heart rate variability (HRV) detects the early subclinical alterations of the autonomic nervous system. Thus, impaired HRV is the earliest subclinical marker of cardiac autonomic neuropathy (CAN) in type 1 diabetes mellitus (T1DM). We aimed to explore the HRV parameters in asymptomatic T1DM patients and compare them with the results obtained in healthy subjects. Potential associations between HRV parameters and the established risk factors for CAN and cardiovascular diseases were also investigated. Seventy T1DM patients (38 ± 12 years, 46 females) and 30 healthy subjects were enrolled into the study. For HRV analysis, beat-to-beat heart rate was recorded for 30 min. The less noisy 5-min segment of the recording was analyzed by Bittium Cardiac Navigator HRV analysis software. Time domain, frequency domain, and nonlinear indices were calculated. Regarding ratio of low to high frequency component (LF/HF), no differences were found between the two populations (p = 0.227). All the further, time domain, frequency domain, and nonlinear HRV indices were significantly lower in T1DM patients (each p < 0.001). In multiple linear models, disease duration remained the only independent predictor of LF/HF ratio (p = 0.019). HbA1c was found to be significant independent predictor of all further time domain (SDNN, p < 0.001; rMSSD, p < 0.001), frequency domain (VLF, p < 0.001; LF, p = 0.002; HF, p = 0.006; Total Power, p = 0.002), and nonlinear indices (SD1, p = 0.006; SD2, p = 0.007), alone, or in combination with other factors, such as age or body mass index. Asymptomatic T1DM patients have significantly reduced overall HRV as compared with healthy subjects, indicating subclinical CAN. Quality of the glycemic control is important determinant of HRV among T1DM patients. This relationship is independent of other risk factors for CAN or cardiovascular diseases.


Cross-reactivity and 1.4-A crystal structure of Malassezia sympodialis thioredoxin (Mala s 13), a member of a new pan-allergen family.
We have identified thioredoxins (Trx) of Malassezia sympodialis, a yeast involved in the pathogenesis of atopic eczema, and of Aspergillus fumigatus, a fungus involved in pulmonary complications, as novel IgE-binding proteins. We show that these Trx, including the human enzyme, represent cross-reactive structures recognized by serum IgE from individuals sensitized to M. sympodialis Trx. Moreover, all three proteins were able to elicit immediate-type allergic skin reactions in sensitized individuals, indicating a humoral immune response based on molecular mimicry. To analyze structural elements involved in these reactions, the three-dimensional structure of M. sympodialis Trx (Mala s 13) has been determined at 1.4-A resolution by x-ray diffraction analysis. The structure was solved by molecular replacement and refined to a crystallographic R factor of 14.0% and a free R factor of 16.8% and shows the typical Trx fold. Mala s 13 shares 45% sequence identity with human Trx and superposition of the solved Mala s 13 structure with those of human Trx reveals a high similarity with a root mean square deviation of 1.11 A for all Calpha atoms. In a detailed analysis of the molecular surface in combination with sequence alignment, we identified conserved solvent-exposed amino acids scattered over the surface in both structures which cluster to patches, thus forming putative conformational B cell epitopes potentially involved in IgE-mediated cross- and autoreactivity.


Tactile sensor is useful for estimating liver hardness and liver fibrosis compared with ultrasonography and computed tomography.
We examined whether conventional ultrasonography (US) and computed tomography (CT) were useful to evaluate liver hardness and hepatic fibrosis by comparing the results with those obtained by a tactile sensor using rats with liver fibrosis. We used 44 Wistar rats in which liver fibrosis was induced by intraperitoneal administration of thioacetamide. The CT and US values of each liver were measured before laparotomy. After laparotomy, a tactile sensor was used to measure liver hardness. We prepared Azan stained sections of each excised liver specimen and calculated the degree of liver fibrosis (HFI: hepatic fibrosis index) by computed color image analysis. The stiffness values and HFI showed a positive correlation (r=0.690, p<0.001), as did the tactile values and HFI (r=0.709, p<0.001).In addition, the stiffness and tactile values correlated positively with each other (r=0.814, p<0.001). There was no correlation between the CT values and HFI, as well as no correlation between the US values and HFI. We confirmed that it was difficult to evaluate liver hardness and HFI by CT or US examination, and considered that, at present, a tactile sensor is useful method for evaluating HFI.


Chromosome 12p deletions in TEL-AML1 childhood acute lymphoblastic leukemia are associated with retrotransposon elements and occur postnatally.
TEL-AML1 (ETV6-RUNX1) is the most common translocation in the childhood leukemias, and is a prenatal mutation in most children. This translocation has been detected at a high rate among newborns ( approximately 1%); therefore, the rate-limiting event for leukemia seems to be secondary mutations. One such frequent mutation in this subtype is partial deletion of chromosome 12p, trans from the translocation. Nine del(12p) breakpoints within six leukemia cases were sequenced to explore the etiology of this genetic event, and most involved cryptic sterile translocations. Twelve of 18 del(12p) parent sequences involved in these breakpoints were located in repeat regions (8 of these in long interspersed nuclear elements). This stands in contrast with TEL-AML1, in which only 21 of 110 previously assessed breakpoints (19%) occur in DNA repeats (P=0.0001). An exploratory assessment of archived neonatal blood cards revealed significantly more long interspersed nuclear element CpG methylations in individuals at birth who were later diagnosed with TEL-AML1 leukemia, compared with individuals who did not contract leukemia (P=0.01). Nontemplate nucleotides were also more frequent in del(12p) than in TEL-AML1 junctions (P=0.004), suggesting formation by terminal deoxynucleotidyl transferase. Assessment of six archived neonatal blood cards indicated that no del(12p) rearrangements backtracked to birth, although two of these patients were previously positive for TEL-AML1 using the same assay with comparable sensitivity. These data are compatible with a two-stage natural history: TEL-AML1 occurs prenatally, and del(12p) occurs postnatally in more mature cells with a structure that suggests the involvement of retrotransposon instability.