Susceptibility of Aedes aegypti and Aedes albopictus to temephos in four study sites in Kuala Lumpur City Center and Selangor State, Malaysia.
Larvae obtained from Taman Samudera (Gombak, Selangor), Kampung Banjar (Gombak, Selangor), Taman Lembah Maju (Cheras, Kuala Lumpur) and Kampung Baru (City centre, Kuala Lumpur) were bioassayed with diagnostic dosage (0.012 mg/L) and operational dosage (1 mg/L) of temephos. All strains of Aedes aegypti and Aedes albopictus showed percentage mortality in the range of 16.00 to 59.05 and 6.4 to 59.50 respectively, after 24 hours. LT50 values for the 6 strains of Ae. aegypti and Ae. albopictus were between 41.25 to 54.42 minutes and 52.67 to 141.76 minutes respectively, and the resistance ratio for both Aedes species were in the range of 0.68 to 1.82 when tested with operational dosage, 1 mg/L temephos. These results indicate that Aedes mosquitoes have developed some degree of resistance. However, complete mortality for all strains were achieved after 24 hours when tested against 1 mg/L temephos.


Generalized length scales for three-dimensional dendritic growth.
The crucial length scale of dendritic growth is the tip radius. Usually it is determined by fitting the data to a theoretical function. We present a method of a generalized tip radius which is entirely based on geometric considerations and is not dependent on an underlying assumption of the shape of the tip. Furthermore the results are stable and the average change of the tip radius between successive images is less than 6%.


Reconstructing uniformly attenuated rotating slant-hole SPECT projection data using the DBH method.
This work applies a previously developed analytical algorithm to the reconstruction problem in a rotating multi-segment slant-hole (RMSSH) SPECT system. The RMSSH collimator has greater detection efficiency than the parallel-hole collimator with comparable spatial resolution at the expense of limited common volume-of-view (CVOV) and is therefore suitable for detecting low-contrast lesions in breast, cardiac and brain imaging. The absorption of gamma photons in both the human breast and brain can be assu- med to follow an exponential rule with a constant attenuation coefficient. In this work, the RMSSH SPECT data of a digital NCAT phantom with breast attachment are modeled as the uniformly attenuated Radon transform of the activity distribution. These data are reconstructed using an analytical algorithm called the DBH method, which is an acronym for the procedure of differentiation backprojection followed by a finite weighted inverse Hilbert transform. The projection data are first differentiated along a specific direction in the projection space and then backprojected to the image space. The result from this first step is equal to a one-dimensional finite weighted Hilbert transform of the object; this transform is then numerically inverted to obtain the reconstructed image. With the limited CVOV of the RMSSH collimator, the detector captures gamma photon emissions from the breast and from parts of the torso. The simulation results show that the DBH method is capable of exactly reconstructing the activity within a well-defined region-of-interest (ROI) within the breast if the activity is confined to the breast or if the activity outside the CVOV is uniformly attenuated for each measured projection, while a conventional filtered backprojection algorithm only reconstructs the high frequency components of the activity function in the same geometry.


[Aspartate kinase from the cyanobacteriium Plectonema boryanum infected with the cyanophage LPP-3].
The effect of development of cyanophage infection on the activity of aspartate kinase of Plectonema boryanum has been studied. It has been determined that activity of aspartate kinase increased during early period of reproduction of cyanophage. It coincided in time with the increase of the level of amino acids of aspartate family. Isoenzymes of aspartate kinase were isolated from the infected cells purified and studied. Expression of viral genome is accompanied with the appearance of four new isoenzymes determined by virus. The revealed aspartate kinases are not subject of regulation by amino acids (the end-products of biosynthesis) according to the principle of feed-back inhibition.


Development of a Brief Patient-Administered Screening Tool for Prescription Opioid Dependence for Primary Care Settings.
To develop a short, patient-administered screening tool that will allow for earlier assessment of prescription opioid dependence (often referred to as addiction) in primary care settings. Cross-sectional analysis (N = 1,134) from the two-year time point of the Pain and Opioids IN Treatment (POINT) cohort was used in the scale development. Participants who completed two-year interviews in the POINT study, a prospective cohort study that followed people with chronic noncancer pain over a five-year period, and who were prescribed strong opioids for a minimum of six weeks at baseline. An advisory committee provided advice on wording and content for screening in primary care settings. Univariate logistic regression identified individual items that were significantly associated with meeting ICD-11 criteria for prescription opioid dependence. Exploratory and confirmatory factor analysis (EFA and CFA) were conducted, and items were reduced to identify a small item set that were discriminative and shared a simple underlying structure. Sixty-four variables associated with ICD-11 criteria for prescription opioid dependence were initially identified. Four rounds of EFA were performed, resulting in five items remaining. CFA identified two possible four-item combinations, with the final combination chosen based on greater item endorsement and the results of goodness-of-fit indices. Addressing prescription opioid dependence is an important part of the global public health challenge surrounding rising opioid-related harm. This study addresses an important initial requisite step to develop a brief screening tool. Further studies are required to validate the tool in clinical settings.


Novel Tretinoin 0.05% Lotion for the Once-Daily Treatment of Moderate-to-Severe Acne Vulgaris in an Asian Population
BACKGROUND: Acne is a common problem among Asian adolescents and adults. Generally, Asian skin is more pigmented, with a higher risk of acne sequelae. Potential for skin irritation and dryness, as well as pigmentary changes are key concerns that can have significant impact on Quality of Life (QoL). The first lotion formulation of tretinoin was developed using novel polymeric emulsion technology to provide an important alternative option to treat acne patients who may be sensitive to the irritant effects of other tretinoin formulations. OBJECTIVE: To evaluate the efficacy, tolerability, and safety of tretinoin 0.05% lotion in treating moderate-to-severe acne in an Asian population. METHODS: Post hoc analysis of two multicenter, randomized, double-blind, vehicle-controlled phase 3 studies. Asian subjects (aged 12 to 48 years, N=69 with 61% female) were randomized (1:1) to receive tretinoin 0.05% lotion or vehicle, once-daily for 12 weeks. Efficacy assessments included changes in baseline inflammatory and noninflammatory lesions and treatment success (at least 2-grade reduction in Evaluator's Global Severity Score [EGSS] and clear/almost clear). Quality of Life (QoL) was assessed using the validated Acne QoL scale. Safety, adverse events (AEs), cutaneous tolerability and hyper- or hypo-pigmentation (using 4-point scales where 0=none and 3=severe) were evaluated. RESULTS: At week 12, mean percent reduction in inflammatory and noninflammatory lesion counts were 58.6% and 51.4% respectively compared with 41.5% and 23.9% with vehicle (P=0.012 for noninflammatory lesions from week 8). Treatment success was achieved by 27.2% of subjects treated with tretinoin 0.05% lotion by week 12. For each Acne QoL domain, changes from baseline achieved with tretinoin 0.05% lotion were statistically significant compared to vehicle. Only five subjects reported any AE; all AEs were mild or moderate and transient. There were no serious AEs (SAEs). There were no treatment-related AEs with tretinoin 0.05% lotion. There were slight transient increases in scaling and burning over the first 4-8 weeks. Mild hyperpigmentation was reported at baseline (mean score, 0.8) and remained mild throughout the study. CONCLUSIONS: Post hoc analysis showed that tretinoin 0.05% lotion was significantly more effective than its vehicle in achieving reductions in noninflammatory acne lesions and improvements in QoL in an Asian population. The novel lotion formulation was well-tolerated, with no treatment-related AEs and no concerns with skin dryness, irritation, or hyperpigmentation. J Drugs Dermatol. 2019;18(9):910-916.


Phthalate esters enhance quinolinate production by inhibiting alpha-amino-beta-carboxymuconate-epsilon-semialdehyde decarboxylase (ACMSD), a key enzyme of the tryptophan pathway.
Tryptophan is metabolized to alpha-amino-beta-carboxymuconate-epsilon-semialdehyde (ACMS) via 3-hydroxyanthranilate (3-HA). ACMS decarboxylase (ACMSD) directs ACMS to acetyl CoA; otherwise ACMS is non-enzymatically converted to quinolinate (QA), leading to the formation of NAD and its degradation products. Thus, ACMSD is a critical enzyme for tryptophan metabolism. Phthalate esters have been suspected of being environmental endocrine disrupters. Because of the structural similarity of phthalate esters with tryptophan metabolites, we examined the effects of phthalate esters on tryptophan metabolism. Phthalate esters containing diets were orally given to rats and the urinary excreted tryptophan metabolites were quantified. Of the phthalate esters with different side chains tested, di(2-ethylhexyl)phthalate (DEHP) and its metabolite, mono(2-ethylhexyl)phthalate (MEHP), most strongly enhanced the production of QA and degradation products of nicotinamide, while 3-HA was unchanged. This pattern of metabolic change led us to assume that these esters lowered ACMSD protein or its activity. Although DEHP could not be tested because of its low solubility, MEHP reversibly inhibited ACMSD from rat liver and mouse kidney, and also the recombinant human enzyme. Correlation between inhibition of ACMSD by phthalate esters with different side chains and urinary excretion of QA supports the notion that phthalate esters perturb tryptophan metabolism by inhibiting ACMSD. Quinolinate is a potential endogenous toxin and has been implicated in the pathogenesis of various disorders. Although toxicity of phthalate esters through accumulation of QA remains to be investigated, they may be detrimental by acting as metabolic disrupters when intake of a tryptophan-rich diet and exposure to phthalate esters occur coincidentally.


Vascular Tissue-Type Plasminogen Activator Promotes Intracranial Aneurysm Formation.
Although the mechanisms that contribute to intracranial aneurysm (IA) formation and rupture are not totally elucidated, inflammation and matrix remodeling are incriminated. Because tPA (tissue-type plasminogen activator) controls both inflammatory and matrix remodeling processes, we hypothesized that tPA could be involved in the pathophysiology of IA. Immunofluorescence analyses of tPA and its main substrate within the aneurysmal wall of murine and human samples were performed. We then compared the formation and rupture of IAs in wild-type, tPA-deficient and type 1 plasminogen activator inhibitor-deficient mice subjected to a model of elastase-induced IA. The specific contribution of vascular versus global tPA was investigated by performing hepatic hydrodynamic transfection of a cDNA encoding for tPA in tPA-deficient mice. The formation and rupture of IAs were monitored by magnetic resonance imaging tracking for 28 days. Immunofluorescence revealed increased expression of tPA within the aneurysmal wall. The number of aneurysms and their symptomatic ruptures were significantly lower in tPA-deficient than in wild-type mice. Conversely, they were higher in plasminogen activator inhibitor-deficient mice. The wild-type phenotype could be restored in tPA-deficient mice by selectively increasing circulating levels of tPA via hepatic hydrodynamic transfection of a cDNA encoding for tPA. Altogether, this preclinical study demonstrates that the tPA present in the blood stream is a key player of the formation of IAs. Thus, tPA should be considered as a possible new target for the prevention of IAs formation and rupture.


Oncogenic bystander radiation effects in Patched heterozygous mouse cerebellum.
The central dogma of radiation biology, that biological effects of ionizing radiation are a direct consequence of DNA damage occurring in irradiated cells, has been challenged by observations that genetic/epigenetic changes occur in unexposed "bystander cells" neighboring directly-hit cells, due to cell-to-cell communication or soluble factors released by irradiated cells. To date, the vast majority of these effects are described in cell-culture systems, while in vivo validation and assessment of biological consequences within an organism remain uncertain. Here, we describe the neonatal mouse cerebellum as an accurate in vivo model to detect, quantify, and mechanistically dissect radiation-bystander responses. DNA double-strand breaks and apoptotic cell death were induced in bystander cerebellum in vivo. Accompanying these genetic events, we report bystander-related tumor induction in cerebellum of radiosensitive Patched-1 (Ptch1) heterozygous mice after x-ray exposure of the remainder of the body. We further show that genetic damage is a critical component of in vivo oncogenic bystander responses, and provide evidence supporting the role of gap-junctional intercellular communication (GJIC) in transmission of bystander signals in the central nervous system (CNS). These results represent the first proof-of-principle that bystander effects are factual in vivo events with carcinogenic potential, and implicate the need for re-evaluation of approaches currently used to estimate radiation-associated health risks.


Real-time pure shift ¹⁵N HSQC of proteins: a real improvement in resolution and sensitivity.
Spectral resolution in proton NMR spectroscopy is reduced by the splitting of resonances into multiplets due to the effect of homonuclear scalar couplings. Although these effects are often hidden in protein NMR spectroscopy by low digital resolution and routine apodization, behind the scenes homonuclear scalar couplings increase spectral overcrowding. The possibilities for biomolecular NMR offered by new pure shift NMR methods are illustrated here. Both resolution and sensitivity are improved, without any increase in experiment time. In these experiments, free induction decays are collected in short bursts of data acquisition, with durations short on the timescale of J-evolution, interspersed with suitable refocusing elements. The net effect is real-time (t 2) broadband homodecoupling, suppressing the multiplet structure caused by proton-proton interactions. The key feature of the refocusing elements is that they discriminate between the resonances of active (observed) and passive (coupling partner) spins. This can be achieved either by using band-selective refocusing or by the BIRD element, in both cases accompanied by a nonselective 180° proton pulse. The latter method selects the active spins based on their one-bond heteronuclear J-coupling to (15)N, while the former selects a region of the (1)H spectrum. Several novel pure shift experiments are presented, and the improvements in resolution and sensitivity they provide are evaluated for representative samples: the N-terminal domain of PGK; ubiquitin; and two mutants of the small antifungal protein PAF. These new experiments, delivering improved sensitivity and resolution, have the potential to replace the current standard HSQC experiments.


Lad, an adapter protein interacting with the SH2 domain of p56lck, is required for T cell activation.
T cell-specific Src family tyrosine kinase, p56lck, plays crucial roles in T cell differentiation, activation, and proliferation. These multiple functions of p56lck are believed to be conducted through the protein-protein interactions with various cellular signaling proteins. To clarify the mechanisms through which p56lck contributes to T cell signaling, we identified the proteins binding to the Src homology 2 (SH2) domain of p56lck through a tyrosine phosphorylation-dependent yeast two-hybrid screening. Subsequent characterization of positive clones revealed the presence of a protein of 366 aa named Lad (Lck-associated adapter protein), which is a potential murine homologue of previously reported TSAd, a T cell-specific adapter protein. Lad contains several protein-protein interaction domains including a zinc-finger motif, an SH2 domain, a proline-rich SH3 binding motif, and several phosphotyrosine sites. Furthermore, Lad was tyrosine phosphorylated and associated with p56lck in vivo and redistributed from cytoplasm to the plasma membrane in a T cell activation-dependent manner. Moreover in T cells, IL-2 promoter activity was enhanced upon coexpression of Lad but was inhibited by the coexpression of antisense Lad RNA. These characteristics of Lad suggest that Lad play an essential role as an adapter protein in p56lck-mediated T cell signaling.


Peginterferon Alfa-2a (40KD) Plus Lamivudine or Entecavir in Children With Immune-Tolerant Chronic Hepatitis B.
Treatment guidelines for chronic hepatitis B (CHB) do not recommend antiviral therapy for patients in the immune-tolerant phase of the disease, which generally occurs in children who acquire hepatitis B virus (HBV) vertically and may last for decades. On the basis of promising results of a pilot study, we conducted a randomized, controlled, multicenter study to evaluate the efficacy and safety of antiviral therapy in children and adolescents with immune-tolerant CHB. Fifty-nine children aged 3 to <18 years hepatitis B e antigen-positive with an HBV DNA titer >20,000 IU/mL and persistently normal alanine aminotransferase levels were randomized to 56 weeks of antiviral therapy with an oral nucleoside analogue [entecavir or lamivudine], combined with subcutaneous peginterferon alfa-2a from week 8, or 80 weeks of untreated observation. The primary efficacy outcome was hepatitis B surface antigen loss 24 weeks post-treatment in the antiviral therapy group or at the end of observation in the control group. Enrollment was terminated after the results of two similar studies showed that similar antiviral regimens were ineffective in children and adults with immune-tolerant CHB. At 24 weeks post-treatment, 1 of 26 patients in the antiviral treatment group experienced HBsAg loss (vs none of 33 patients in the control group). No serious treatment-related adverse events were reported, and no patients discontinued treatment because of adverse events. The antiviral regimen evaluated in this trial had an acceptable tolerability profile, but was ineffective in children and adolescents with immune-tolerant CHB.


Atomic resolution description of the interaction between the nucleoprotein and phosphoprotein of Hendra virus.
Hendra virus (HeV) is a recently emerged severe human pathogen that belongs to the Henipavirus genus within the Paramyxoviridae family. The HeV genome is encapsidated by the nucleoprotein (N) within a helical nucleocapsid. Recruitment of the viral polymerase onto the nucleocapsid template relies on the interaction between the C-terminal domain, N(TAIL), of N and the C-terminal X domain, XD, of the polymerase co-factor phosphoprotein (P). Here, we provide an atomic resolution description of the intrinsically disordered N(TAIL) domain in its isolated state and in intact nucleocapsids using nuclear magnetic resonance (NMR) spectroscopy. Using electron microscopy, we show that HeV nucleocapsids form herringbone-like structures typical of paramyxoviruses. We also report the crystal structure of XD of P that consists of a three-helix bundle. We study the interaction between N(TAIL) and XD using NMR titration experiments and provide a detailed mapping of the reciprocal binding sites. We show that the interaction is accompanied by α-helical folding of the molecular recognition element of N(TAIL) upon binding to a hydrophobic patch on the surface of XD. Finally, using solution NMR, we investigate the interaction between intact nucleocapsids and XD. Our results indicate that monomeric XD binds to N(TAIL) without triggering an additional unwinding of the nucleocapsid template. The present results provide a structural description at the atomic level of the protein-protein interactions required for transcription and replication of HeV, and the first direct observation of the interaction between the X domain of P and intact nucleocapsids in Paramyxoviridae.


Frequent deletion in the methylthioadenosine phosphorylase gene in T-cell acute lymphoblastic leukemia: strategies for enzyme-targeted therapy.
Methylthioadenosine phosphorylase (MTAP), an enzyme essential for the salvage of adenine and methionine, is deficient in a variety of cancers, including acute lymphoblastic leukemia (ALL). Because the MTAP gene is located adjacent to the tumor-suppressor gene p16 on chromosome 9p21 and more than 60% of T-cell ALL (T-ALL) patients have deletion in the p16 gene, we examined the status of the MTAP gene in T-ALL patients. Quantitative polymerase chain reaction amplification of exon 8 of MTAP showed a deletion in 16 of 48 (33.3%) patients at diagnosis and in 13 of 33 (39.4%) patients at relapse. Southern blot analysis showed that, in addition to deletion of the entire MTAP gene, a common break point was between exons 4 and 5, resulting in deletion of exons 5 through 8. The finding of frequent deficiency of MTAP in T-ALL offers the possibility of an enzyme targeted therapy for T-ALL. MTAP(-) T-ALL-derived cell line, CEM cells were very sensitive to methionine deprivation, with cell viability at 50% of control as early as 48 hours after methionine deprivation. In contrast, methionine deprivation had little effect on the viability of normal lymphocytes or on their proliferative response to phytohemagglutinin. Alanosine, an inhibitor of AMP synthesis, inhibited the growth of both MTAP(+) (Molt-4 and Molt-16) and MTAP(-) (CEM and HSB2) cell lines. However, the addition of methylthioadenosine, the substrate of MTAP, protected the MTAP(+) cells but not the MTAP(-) cells from alanosine toxicity. These findings suggest the possibility of targeting MTAP for selective therapy of T-ALL.


Macrophages and HIV infection: therapeutical approaches toward this strategic virus reservoir.
Cells of macrophage lineage represent a key target of human immunodeficiency virus (HIV) in addition to CD4-lymphocytes. The absolute number of infected macrophages in the body is relatively low compared to CD4-lymphocytes. Nevertheless, the peculiar dynamics of HIV replication in macrophages, their long-term survival after HIV infection, and their ability to spread virus particles to bystander CD4-lymphocytes, make evident their substantial contribution to the pathogenesis of HIV infection. In addition, infected macrophages are able to recruit and activate CD4-lymphocytes through the production of both chemokines and virus proteins (such as nef). In addition, the activation of the oxidative pathway in HIV-infected macrophages may lead to apoptotic death of bystander, not-infected cells. Finally, macrophages are the most important target of HIV in the central nervous system. The alteration of neuronal metabolism induced by infected macrophages plays a crucial role in the pathogenesis of HIV-related encephalopathy. Taken together, these results strongly support the clinical relevance of therapeutic strategies able to interfere with HIV replication in macrophages. In vitro data show the potent efficacy of all nucleoside analogues inhibitors of HIV-reverse transcriptase in macrophages. Nevertheless, the limited penetration of some of these compounds in sequestered districts, coupled with the scarce phosphorylation ability of macrophages, suggests that nucleoside analogues carrying preformed phosphate groups may have a potential role against HIV replication in macrophages. This hypothesis is supported by the great anti-HIV activity of tenofovir and other acyclic nucleoside phosphonates in macrophages that may provide a rationale for the remarkable efficacy of tenofovir in HIV-infected patients. Non-nucleoside reverse transcriptase inhibitors (NNRTI) do not affect HIV-DNA chain termination, and for this reason their antiviral activity in macrophages is similar to that found in CD4-lymphocytes. Interestingly, protease inhibitors (PIs), acting at post-integrational stages of virus replication, are the only drugs able to interfere with virus production and release from macrophages with established and persistent HIV infection (chronically-infected cells). Since this effect is achieved at concentrations and doses higher than those effective in de-novo infected CD4-lymphocytes, it is possible that lack of adherence to therapy, and/or suboptimal dosage leading to insufficient concentrations of PIs may cause a resumption of virus replication from chronically-infected macrophages, ultimately resulting in therapeutic failure. For all these reasons, therapeutic strategies aimed to achieve the greatest and longest control of HIV replication should inhibit HIV not only in CD4-lymphocytes, but also in macrophages. Testing new and promising antiviral compounds in such cells may provide crucial hints about their efficacy in patients infected by HIV.


First-in-Human Phase 1 Study To Assess Safety, Tolerability, and Pharmacokinetics of a Novel Antifungal Drug, VL-2397, in Healthy Adults.
VL-2397 is an antifungal drug with a novel mechanism of action, rapid fungicidal in vitro activity, and potent in vivo activity against Aspergillus fumigatus, including azole-resistant strains. VL2397-101, a phase 1 first-in-human, randomized, double-blind, placebo-controlled dose-escalation study, was conducted in healthy adults to determine the safety, tolerability, and pharmacokinetics (PK) of single and multiple ascending intravenous (i.v.) doses of VL-2397. All dosing cohorts were fully enrolled; all subjects completed the safety follow-up. A safety committee reviewed the safety data for each dosing cohort prior to recommending the initiation of each subsequent cohort. No serious adverse events (SAEs) occurred; the majority of treatment-emergent adverse events (TEAEs) were mild and self-limited. The most common drug-related TEAEs were infusion site reactions. No clinically concerning trends were noted in vital signs, electrocardiograms, physical examinations, or safety laboratory results. Following single infusions of VL-2397, the overall and maximum exposures rose less than proportionally with increasing doses from 3 mg to 1,200 mg as indicated by area under the concentration-time curve over 24 h (AUC24) and maximum concentration (Cmax). No signs of VL-2397 accumulation were observed following i.v. infusions of 300, 600, and 1,200 mg every 24 h (q24h) for 7 days. Renal elimination played a major role in total body clearance, with up to 47% of unmetabolized drug in urine 24 h after administration at single doses of >30 mg. Overall, VL-2397 dosing in the study appeared to be safe and well tolerated in the healthy subjects. The safety profile, consistent PK, and lack of drug accumulation support further development of VL-2397 in patients with invasive aspergillosis.


Reduced risk of severe COVID-19 in more than 1.4 million elderly people aged 75 years and older vaccinated with mRNA-based vaccines.
Randomized clinical trials have shown mRNA-based vaccines to be 92-95% effective to prevent COVID-19 in adults. We aimed to estimate the impact of vaccination on the risk of severe COVID-19 (requiring hospitalization) in elderly people. Each 1,422,461 vaccinated subject aged 75 or older was matched to two unvaccinated subjects of same age, sex, administrative region, and type of residence. They were followed from date of first injection between 27 December 2020 and 24 February 2021 to 20 March 2021 for COVID-19 hospitalization. Mean age was 82.4 years (SD, 5.7) and median follow-up was 38 days [IQR, 17-54]. Adjusted Hazard Ratio for COVID-19 hospitalization from day 7 after the second dose was estimated at 0.14 (95% confidence interval, 0.11-0.17), i.e. an estimated 86% risk reduction in people aged 75 and older, highlighting the major impact of mRNA vaccination on reducing the risk of COVID-19 among elderly people.


High-power distributed Bragg reflector ridge-waveguide diode laser with very small spectral linewidth.
We manufactured and investigated distributed Bragg reflector ridge-waveguide diode lasers having sixth-order surface gratings and an emission wavelength around 974 nm. The single-mode output power of the lasers with a total length of 4 mm exceeded 1 W. A very small spectral linewidth of 1.4 MHz (3 dB) consisting of a Lorentzian part of 146 kHz and a Gaussian part of 1308 MHz was measured using a self-delayed heterodyne measurement technique.


A randomized pilot trial of a mobile delivered brief motivational interviewing and behavioral economic alcohol intervention for emerging adults.
Mobile health (mHealth) interventions show potential to broaden the reach of efficacious alcohol brief motivational interventions (BMIs). However, efficacy is mixed and may be limited by low participant attention and engagement. The present study examined the feasibility, acceptability, and preliminary efficacy of a live text-message delivered BMI in a pilot randomized clinical trial. Participants were 66 college students (63.6% women; 61.9% White; Mage = 19.95, SD = 1.66) reporting an average of 11.88 (SD = 8.74) drinks per week, 4.42 (SD = 3.59) heavy drinking episodes (HDEs), and 8.44 (SD = 5.62) alcohol-related problems in the past month. Participants were randomized to receive either (a) education or (b) an alcohol BMI plus behavioral economic substance-free activity session (SFAS), each followed by 4 weeks of mini sessions. All sessions were administered via live text-message. Participants completed assessments postintervention (after the 4th mini session) and at 3-month follow-up. 90.9% completed both initial full-length sessions and at least two of the four mini sessions with 87.9% retention at 3-month follow-up. Participants found the interventions useful, interesting, relevant, and effective, with no between-group differences. There were no statistically significant group differences in drinks per week or alcohol-related problems at follow-up, but BMI + SFAS participants reported fewer past-month HDEs than those who received education. Live text-messaging to deliver the BMI + SFAS is feasible and well-received. The preliminary efficacy results should be interpreted cautiously due to the small sample size but support further investigation. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


Production of Protein Hydrolysate from Quinoa (Chenopodium quinoa Willd.): Economic and Experimental Evaluation of Two Pretreatments Using Supercritical Fluids' Extraction and Conventional Solvent Extraction.
Supercritical fluids' extraction (SFE) and conventional solvent extraction (CSE) for defatting of quinoa flour as pretreatments to produce the quinoa protein hydrolysate (QPH) were studied. The objective was to extract the oil and separate the phenolic compounds (PC) and the defatted quinoa flour for subsequent quinoa protein extraction and enzymatic hydrolysis. The oil extraction yield (OEY), total flavonoid content (TFC), and QPH yield were compared. SuperPro Designer 9.0® software was used to estimate the cost of manufacturing (COM), productivity, and net present value (NPV) on laboratory and industrial scales. SFE allows higher OEY and separation of PC. The SFE oil showed a higher OEY (99.70%), higher antioxidant activity (34.28 mg GAE/100 g), higher QPH yield (197.12%), lower COM (US$ 90.10/kg), and higher NPV (US$ 205,006,000) as compared to CSE (with 77.59%, 160.52%, US$ 109.29/kg, and US$ 28,159,000, respectively). The sensitivity analysis showed that the sale of by-products improves the economic results: at the industrial scale, no significant differences were found, and both processes are economically feasible. However, results indicate that SFE allows the recovery of an oil and QPH of better nutritional quality and a high level of purity-free organic solvents for further health and nutraceutical uses.


NOX4 links metabolic regulation in pancreatic cancer to endoplasmic reticulum redox vulnerability and dependence on PRDX4.
There is an urgent need to identify vulnerabilities in pancreatic ductal adenocarcinoma (PDAC). PDAC cells acquire metabolic changes that augment NADPH production and cytosolic redox homeostasis. Here, we show that high NADPH levels drive activity of NADPH oxidase 4 (NOX4) expressed in the endoplasmic reticulum (ER) membrane. NOX4 produces H2O2 metabolized by peroxiredoxin 4 (PRDX4) in the ER lumen. Using functional genomics and subsequent in vitro and in vivo validations, we find that PDAC cell lines with high NADPH levels are dependent on PRDX4 for their growth and survival. PRDX4 addiction is associated with increased reactive oxygen species, a DNA-PKcs-governed DNA damage response and radiosensitivity, which can be rescued by depletion of NOX4 or NADPH. Hence, this study has identified NOX4 as a protein that paradoxically converts the reducing power of the cytosol to an ER-specific oxidative stress vulnerability in PDAC that may be therapeutically exploited by targeting PRDX4.


The NADPH Oxidases DUOX1 and NOX2 Play Distinct Roles in Redox Regulation of Epidermal Growth Factor Receptor Signaling.
The epidermal growth factor receptor (EGFR) plays a critical role in regulating airway epithelial homeostasis and responses to injury. Activation of EGFR is regulated by redox-dependent processes involving reversible cysteine oxidation by reactive oxygen species (ROS) and involves both ligand-dependent and -independent mechanisms, but the precise source(s) of ROS and the molecular mechanisms that control tyrosine kinase activity are incompletely understood. Here, we demonstrate that stimulation of EGFR activation by ATP in airway epithelial cells is closely associated with dynamic reversible oxidation of cysteine residues via sequential sulfenylation and S-glutathionylation within EGFR and the non-receptor-tyrosine kinase Src. Moreover, the intrinsic kinase activity of recombinant Src or EGFR was in both cases enhanced by H2O2 but not by GSSG, indicating that the intermediate sulfenylation is the activating modification. H2O2-induced increase in EGFR tyrosine kinase activity was not observed with the C797S variant, confirming Cys-797 as the redox-sensitive cysteine residue that regulates kinase activity. Redox-dependent regulation of EGFR activation in airway epithelial cells was found to strongly depend on activation of either the NADPH oxidase DUOX1 or the homolog NOX2, depending on the activation mechanism. Whereas DUOX1 and Src play a primary role in EGFR transactivation by wound-derived signals such as ATP, direct ligand-dependent EGFR activation primarily involves NOX2 with a secondary role for DUOX1 and Src. Collectively, our findings establish that redox-dependent EGFR kinase activation involves a dynamic and reversible cysteine oxidation mechanism and that this activation mechanism variably involves DUOX1 and NOX2.


Changes in cognitive domains during three years in patients with Alzheimer's disease treated with donepezil.
The objective was to identify separate cognitive domains in the standard assessment tools (MMSE, ADAS-Cog) and analyze the process of decline within domains during three years in Alzheimer's disease (AD) patients with donepezil treatment. AD patients (n = 421) were recruited from a clinical multi-centre study program in Sweden. Patients were assessed every six months during three years. All patients received donepezil starting directly after study entry. After dropouts, 158 patients remained for analyses over three years. Data for the other patients were analysed until they dropped out (4 groups based on length in study). Factor analyses of all items suggested that there were three intercorrelated factors: a General, a Memory and a Spatial factor for which we constructed corresponding domains. Overall there was a cognitive improvement at six months followed by a linear drop over time for the three domains. Some group and domain differences were identified. Patients who remained longer in the study had better initial performance and a slower deterioration rate. The early dropouts showed no improvement at six months and many dropped out due to side effects. The other groups displayed a performance improvement at six months that was less pronounced in the Memory domain. Before dropping out, deterioration accelerated, particularly in the Spatial domain. The course of illness in the three domains was heterogeneous among the patients. We were not able to identify any clinically relevant correlates of this heterogeneity. As an aid we constructed three algorithms corresponding to the cognitive domains, which can be used to characterize patients initially, identify rapid decliners and follow the course of the disease.


High-Spin Mn(V)-Oxo Intermediate in Nonheme Manganese Complex-Catalyzed Alkane Hydroxylation Reaction: Experimental and Theoretical Approach.
Mononuclear nonheme manganese complexes are highly efficient catalysts in the catalytic oxidation of hydrocarbons by hydrogen peroxide in the presence of carboxylic acids. Although high-valent Mn(V)-oxo complexes have been proposed as the active oxidants that afford high regio-, stereo-, and enantioselectivities in the catalytic oxidation reactions, the importance of the spin state (e.g., S = 0 or 1) of the proposed Mn(V)-oxo species is an area that requires further study. In the present study, we have theoretically demonstrated that a mononuclear nonheme Mn(V)-oxo species with an S = 1 ground spin state is the active oxidant that effects the stereo- and enantioselective alkane hydroxylation reaction; it is noted that synthetic octahedral Mn(V)-oxo complexes, characterized spectroscopically and/or structurally, possess an S = 0 spin state and are sluggish oxidants. In an experimental approach, we have investigated the catalytic hydroxylation of alkanes by a mononuclear nonheme Mn(II) complex, [(S-PMB)MnII]2+, and H2O2 in the presence of carboxylic acids; alcohol is the major product with high stereo- and enantioselectivities. A synthetic Mn(IV)-oxo complex, [(S-PMB)MnIV(O)]2+, is inactive in C-H bond activation reactions, ruling out the Mn(IV)-oxo species as an active oxidant. DFT calculations have shown that a Mn(V)-oxo species with an S = 1 spin state, [(S-PMB)MnV(O)(OAc)]2+, is highly reactive and capable of oxygenating the C-H bond via oxygen rebound mechanism; we propose that the triplet spin state of the Mn(V)-oxo species results from the consequence of breaking the equatorial symmetry due to the binding of an equatorial oxygen from an acetate ligand. Thus, the present study reports that, different from the previously reported S = 0 Mn(V)-oxo species, Mn(V)-oxo species with a triplet ground spin state are highly reactive oxidants that are responsible for the regio-, stereo-, and enantioselectivities in the catalytic hydroxylation of alkanes by mononuclear nonheme manganese complexes and terminal oxidants.


MYC regulates ribosome biogenesis and mitochondrial gene expression programs through its interaction with host cell factor-1.
The oncoprotein transcription factor MYC is a major driver of malignancy and a highly validated but challenging target for the development of anticancer therapies. Novel strategies to inhibit MYC may come from understanding the co-factors it uses to drive pro-tumorigenic gene expression programs, providing their role in MYC activity is understood. Here we interrogate how one MYC co-factor, host cell factor (HCF)-1, contributes to MYC activity in a human Burkitt lymphoma setting. We identify genes connected to mitochondrial function and ribosome biogenesis as direct MYC/HCF-1 targets and demonstrate how modulation of the MYC-HCF-1 interaction influences cell growth, metabolite profiles, global gene expression patterns, and tumor growth in vivo. This work defines HCF-1 as a critical MYC co-factor, places the MYC-HCF-1 interaction in biological context, and highlights HCF-1 as a focal point for development of novel anti-MYC therapies.


Prevalence of and gene regulatory constraints on transcriptional adaptation in single cells.
Cells and tissues have a remarkable ability to adapt to genetic perturbations via a variety of molecular mechanisms. Nonsense-induced transcriptional compensation, a form of transcriptional adaptation, has recently emerged as one such mechanism, in which nonsense mutations in a gene can trigger upregulation of related genes, possibly conferring robustness at cellular and organismal levels. However, beyond a handful of developmental contexts and curated sets of genes, to date, no comprehensive genome-wide investigation of this behavior has been undertaken for mammalian cell types and contexts. Moreover, how the regulatory-level effects of inherently stochastic compensatory gene networks contribute to phenotypic penetrance in single cells remains unclear. Here we combine computational analysis of existing datasets with stochastic mathematical modeling and machine learning to uncover the widespread prevalence of transcriptional adaptation in mammalian systems and the diverse single-cell manifestations of minimal compensatory gene networks. Regulon gene expression analysis of a pooled single-cell genetic perturbation dataset recapitulates important model predictions. Our integrative approach uncovers several putative hits-genes demonstrating possible transcriptional adaptation-to follow up on experimentally, and provides a formal quantitative framework to test and refine models of transcriptional adaptation.


Efficacy of a motilin receptor agonist (ABT-229) for the treatment of gastro-oesophageal reflux disease.
ABT-229 is a potent motilin agonist without significant antibiotic activity. It has been shown to improve gastric emptying in humans and to increase lower oesophageal sphincter pressure in cats. To assess the efficacy of four different doses of ABT-229 (1.25 mg, 2.5 mg, 5 mg, 10 mg b.d.) compared to placebo in the treatment of gastro-oesophageal reflux disease, and to determine its safety in patients with gastro-oesophageal reflux disease. In a double-blind, multicentre study, 324 patients with heartburn were randomized to receive four different doses of ABT-229 or placebo for 8 weeks. The efficacy was evaluated by Patient Symptom Questionnaire, daily diary, endoscopy and global evaluation of efficacy. There were no statistically significant improvement scores for any of the ABT-229 treatment groups vs. the placebo group in any of the efficacy parameters. Reflux symptom scores were significantly worse after treatment in the dyspeptic group. ABT-229 appeared to be well tolerated and safe in total daily doses up to 20 mg. ABT-229 appears to have limited, if any, clinical utility in the treatment of gastro-oesophageal reflux disease.


Bacillus thermoglucosidius alpha-glucosidase. Temperature dependence of activity and stability.
A p-nitrophenyl-alpha-D-glucopyranoside-hydrolyzing alpha-glucosidase from an obligate thermophile, Bacillus thermoglucosidius KP 1006, gave a triphasic relationship at pH 6.8 in the van't Hoff plot of Km, in the Arrhenius plot of the first order rate constant of inactivation with 0.1% sodium dodecyl sulfate, and in the logarithmic plot of the maximal fluorescence intensity at 346 nm versus reciprocal of temperature. The respective plots exhibited two breaks at 40 and 61 degrees C, 43 and 62 degrees C, and 40 and 61 degrees C. However, the Arrhenius plot of the molecular activity at pH 6.8 had a single discontinuity at 64 degrees C. These findings, together with thermodynamic quantities for the enzyme, suggest that the thermal conformation changes in the enzyme protein occur around 40--43 degrees C and 61--64 degrees C. The Arrhenius plot of the rate constant of heat inactivation at pH 6.8 was bent at 73 degrees C. Thermodynamic data indicate that the enzyme is transformed from a heat stable form into a heat unstable form at 73 degrees C with temperature elevation. The critical points localized near the minimum, optimum, and maximum temperatures (40, 60, and 72 degrees C) of the cell growth, respectively.


High resolution approach to the native state ensemble kinetics and thermodynamics.
Many biologically interesting functions such as allosteric switching or protein-ligand binding are determined by the kinetics and mechanisms of transitions between various conformational substates of the native basin of globular proteins. To advance our understanding of these processes, we constructed a two-dimensional free energy surface (FES) of the native basin of a small globular protein, Trp-cage. The corresponding order parameters were defined using two native substructures of Trp-cage. These calculations were based on extensive explicit water all-atom molecular dynamics simulations. Using the obtained two-dimensional FES, we studied the transition kinetics between two Trp-cage conformations, finding that switching process shows a borderline behavior between diffusive and weakly-activated dynamics. The transition is well-characterized kinetically as a biexponential process. We also introduced a new one-dimensional reaction coordinate for the conformational transition, finding reasonable qualitative agreement with the two-dimensional kinetics results. We investigated the distribution of all the 38 native nuclear magnetic resonance structures on the obtained FES, analyzing interactions that stabilize specific low-energy conformations. Finally, we constructed a FES for the same system but with simple dielectric model of water instead of explicit water, finding that the results were surprisingly similar in a small region centered on the native conformations. The dissimilarities between the explicit and implicit model on the larger-scale point to the important role of water in mediating interactions between amino acid residues.


Exploring the potential of the model cyanobacterium Synechocystis PCC 6803 for the photosynthetic production of various high-value terpenes.
The robust model cyanobacterium Synechocystis PCC 6803 is increasingly explored for its potential to use solar energy, water and atmospheric CO2 for the carbon-neutral production of terpenes, the high-value chemicals that can be used for the production of drugs, flavors, fragrances and biofuels. However, as terpenes are chemically diverse, it is extremely difficult to predict whether Synechocystis is a suitable chassis for the photosynthetic production of various terpenes or only a few of them. We have performed the first-time engineering and comparative analysis of the best-studied cyanobacterium Synechocystis PCC 6803 for the photosynthetic production of five chemically diverse high-value terpenes: two monoterpenes (C10H16) limonene (cyclic molecule) and pinene (bicyclic), and three sesquiterpenes (C15H24) bisabolene (cyclic), farnesene (linear) and santalene (cyclic). All terpene producers appeared to grow well and to be genetically stable, as shown by the absence of changes in their production levels during the 5-9-month periods of their sub-cultivation under photoautotrophic conditions). We also found that Synechocystis PCC 6803 can efficiently and stably produce farnesene and santalene, which had never been produced before by this model organism or any other cyanobacteria, respectively. Similar production levels were observed for cells growing on nitrate (the standard nitrogen source for cyanobacteria) or urea (cheaper than nitrate). Furthermore, higher levels of farnesene were produced by cloning the heterologous farnesene synthase gene in a RSF1010-derived replicating plasmid as compared to the well-used slr0168 neutral cloning site of the chromosome. Altogether, the present results indicate that Synechocystis PCC 6803 is better suited to produce sesquiterpenes (particularly farnesene, the most highly produced terpene of this study) than monoterpenes (especially pinene).


Neural basis for improgan antinociception.
Improgan, the prototype compound of a novel class of non-opioid analgesic drugs derived from histamine antagonists, attenuates thermal and mechanical nociception in rodents following intracerebroventricular (i.c.v.) administration. Improgan does not bind to known opioid, histamine or cannabinoid receptors, and its molecular target has not been identified. It is known however, that improgan acts directly in the periaqueductal gray and the rostral ventromedial medulla to produce its antinociceptive effects, and that inactivation of the rostral ventromedial medulla prevents the antinociceptive effect of improgan given i.c.v. Here we used in vivo single-cell recording in lightly anesthetized rats to show that improgan engages pain-modulating neurons in the medulla to produce antinociception. Following improgan administration, OFF-cells, which inhibit nociception, became continuously active and no longer paused during noxious stimulation. The increase in OFF-cell firing does not represent a non-specific neuroexcitant effect of this drug, since ON-cell discharge, associated with net nociceptive facilitation, was depressed. NEUTRAL-cell firing was unaffected by improgan. The net response of rostral ventromedial medulla (RVM) neurons to improgan is thus comparable to that evoked by mu-opioids and cannabinoids, well known RVM-active analgesic drugs. This common basis for improgan, opioid, and cannabinoid antinociception in the RVM supports the idea that improgan functions as a specific analgesic agent.


[Sudden death in patients with intraventricular conduction disorders in myocardial infarct].
Examination of 900 patients with acute myocardial infarction was carried out, in 110 of them disorders of intraventricular conduction were revealed. Atrioventricular block of grade III-IV developed in 13 patients within the first 3 days of the disease. Nine patients died: asystolia developed in 4 of them, progressive cardiac insufficiency in 3, cardiogenic shock in 1, paroxysm of ventricular tachycardia followed by ventricular fibrillation in 1 patient.


Guidelines for the performance of fusion procedures for degenerative disease of the lumbar spine. Part 17: bone growth stimulators and lumbar fusion.
There have been a number of randomized studies supporting the use of ES for the promotion of bone healing following lumbar fusion. All of the published studies have methodological flaws that prevent the studies from providing Class I medical evidence. There is, however, Class II and III evidence to support the use of direct current stimulation or CCS for enhancing fusion rates in high-risk patients undergoing lumbar PLF. A beneficial effect on fusion rates in patients not at "high risk" has not been convincingly demonstrated, nor has an effect been shown for these modalities in patients treated with interbody fusion. There is limited evidence both for and against the use of PEMFS for enhancing fusion rates following PLE Class II and III medical evidence supports the use of PEMFS for promoting arthrodesis following interbody fusion. Although some studies have purported to demonstrate functional improvement in some patient subgroups, other studies have not detected differences. All of the reviewed studies are significantly flawed by the use of a four-point patient satisfaction scale as the primary outcome measure. This outcome measure is not validated. Because of the use of this flawed outcome measure and because of the conflicting results reported in the better-designed studies that assess functional outcome, there is no consistent medical evidence to support or refute use of these devices for improving patient outcomes.


Identification of a homeologous chromosome pair by in situ DNA hybridization to ribosomal RNA loci in meiotic chromosomes of cotton (Gossypium hirsutum).
In situ DNA hybridization with 18S-28S and 5S ribosomal DNA probes was used to map 18S-28S nucleolar organizers and tandem 5S repeats to meiotic chromosomes of cotton (Gossypium hirsutum L.). Mapping was performed by correlating hybridization sites to particular positions in translocation quadrivalents. Arm assignment required translocation quadrivalents with at least one interstitial chiasma and sufficient distance between the hybridization site and the centromere. We had previously localized a major 18S-28S site to the short arm of chromosome 9; here we mapped two additional major 18S-28S sites to the short arm of chromosome 16 and the left arm of chromosome 23. We also identified and mapped a minor 18S-28S site to the short arm of chromosome 7. Two 5S sites of unequal size were identified, the larger one near the centromere of chromosome 9 and the smaller one near the centromere of chromosome 23. Synteny of 5S and 18S-28S sites indicated homeology of chromosomes 9 and 23, while positions of the other two 18S-28S sites supplement genetic evidence that chromosomes 7 and 16 are homeologous.


Gestational diabetes, comparison of women diagnosed in second and third trimester of pregnancy with non GDM women: Analysis of a cohort study.
Pregnant women are normally screened for Gestational diabetes (GDM) at week 24 of pregnancy. However some women develop the disease later on their pregnancies. No study has analyzed women developing GDM later in pregnancy. To analyze data on a cohort study and compare women diagnosed with GDM in second and third trimester of pregnancy with women without GDM. GDM women diagnosed during their first two trimesters of pregnancy were older (p = 0.0008) and had higher body mass index (BMI) (p = 0.0007) than non GDM women. However, the only risk factor in women diagnosed in their third trimester of pregnancy was having first degree relatives with type 2 DM and this was independent of age and BMI (OR of 2.7, 95% CI 1.2 - 6.0). Women who develop GDM in their second trimester of pregnancy have known risk factors for diabetes mellitus such as age and higher BMI, however, the only recognised risk factor between non GDM women and women developing GDM late in pregnancy is family history of type 2 DM. Two populations of GDM may exist and future studies should focus on analysing short and long term complications of these women to support the need to diagnosed and treat them all.


Plasma cortisol and adrenocorticotropin in appropriate and small for gestational age fetuses.
Plasma cortisol and adrenocorticotropin (ACTH) were measured in fetal blood samples obtained by cordocentesis from 61 appropriate for gestational age (AGA) and in 41 small for gestational age (SGA) fetuses at 18-38 weeks gestation. Fetal plasma cortisol (mean 74 nmol/l) did not change but plasma ACTH increased between 18 and 36 weeks gestation and the two were not significantly correlated. Plasma cortisol in cord blood samples after vaginal delivery at term (mean 305 nmol/l) was higher than in samples at elective cesarean section (mean 151 nmol/l), and there was a significant correlation between fetal and maternal levels. The findings suggest that in human fetuses a late gestational rise in plasma cortisol may not be necessary for organ maturation and that the high fetal plasma cortisol in spontaneous labor at term is probably the result rather than the cause of labor. In SGA fetuses, plasma cortisol was higher and ACTH lower than in AGA fetuses, and plasma cortisol was inversely correlated to fetal hypoglycemia, suggesting that in the chronically hypoglycemic SGA fetus the fetal pituitary is under negative inhibition.


Structures of Escherichia coli NAD synthetase with substrates and products reveal mechanistic rearrangements.
Nicotinamide adenine dinucleotide synthetases (NADS) catalyze the amidation of nicotinic acid adenine dinucleotide (NAAD) to yield the enzyme cofactor nicotinamide adenine dinucleotide (NAD). Here we describe the crystal structures of the ammonia-dependent homodimeric NADS from Escherichia coli alone and in complex with natural substrates and with the reaction product NAD. The structures disclosed two NAAD/NAD binding sites at the dimer interface and an adenosine triphosphate (ATP) binding site within each subunit. Comparison with the Bacillus subtilis NADS showed pronounced chemical differences in the NAAD/NAD binding sites and less prominent differences in the ATP binding pockets. In addition, the E. coli NADS structures revealed unexpected dynamical rearrangements in the NAAD/NAD binding pocket upon NAAD-to-NAD conversion, which define a catalysis state and a substrate/product exchange state. The two states are adopted by concerted movement of the nicotinysyl moieties of NAAD and NAD, Phe-170, and residues 224-228, which may be triggered by differential coordination of a magnesium ion to NAAD and NAD. Phylogenetic structure comparisons suggest that the present results are relevant for designing species-specific antibiotics.


Risk factors associated with severe viral croup in hospitalised Malaysian children.
To determine the risk factors associated with severe viral croup in hospitalised Malaysian children. The medical records of children aged less than five years admitted with a diagnosis of viral croup between 1994 and 1999 were reviewed. Severe viral croup was diagnosed in children who had stridor at rest with marked recession associated with central cyanosis or altered level of consciousness. Multivariate logistic regression analysis was performed to identify risk factors associated with severe viral croup. Eighteen (14.7%) of 122 children with viral croup were severe. These children were older (mean age 16.8 +/- 7.2 vs 12.6 +/- 6.6 months, p = 0.01) and had a shorter duration of illness prior to admission (1.7 +/- 0.7 days vs 2.3 +/- 1.4 days, p = 0.03). Age between 12 and 24 months (OR 3.8 95% Cl 1.3, 12.7, p = 0.02) and fever (OR 5.7 95% Cl 2.9, 15.6, p = 0.02) were the only risk factors associated with severe viral croup after multivariate logistic regression analysis. Only three children or 2.5 per 100 children admitted with viral croup required ventilation. Only a small number of children admitted particularly those between 12 to 24 months with fever developed severe viral croup. Recognition of these risk factors provides a guide in selecting children who will most likely benefit from steroid therapy. The overall outcome was nonetheless favourable.


[Pharmacokinetics of domestic actoprotector drug Metaprot in healthy volunteers].
Pharmacokinetics of the actoprotector Metaprot, an original Russian drug, has been studied in a group of healthy adult volunteers. Metaprot in capsules was administrated orally as a single dose of 250 mg. The concentration of the active substance (ethylthiobenzimidazole) in the blood serum was determined by high-performance liquid chromatography (HPLC) with UV detection. The pharmacokinetic parameters were calculated by the model-independent method. The peak concentration of ethylthiobenzimidazole in plasma was Cmax = 0.91 +/- 1.05 microg/ml and the average time to peak concentration was t(max) = 1.06 +/- 0.16 h. A polymodal character of the distribution of pharmacokinetic parameters in the test group was revealed.


Differential conductance and quantum interference in Kondo systems.
We present a large-N theory for the differential conductance, dI/dV, in Kondo systems measured via scanning tunneling spectroscopy. We demonstrate that quantum interference between tunneling processes into the conduction band and into the magnetic f-electron states is crucial in determining the experimental Fano line shape of dI/dV. This allows one to uniquely extract the Kondo coupling and the ratio of the tunneling amplitudes from the experimental dI/dV curve. Finally, we show that dI/dV directly reflects the strength of the antiferromagnetic interaction in Kondo lattice systems.


Immunosuppression in Glioblastoma: Current Understanding and Therapeutic Implications.
Glioblastoma (GBM) is the most common primary brain tumor in adults an carries and carries a terrible prognosis. The current regiment of surgical resection, radiation, and chemotherapy has remained largely unchanged in recent years as new therapeutic approaches have struggled to demonstrate benefit. One of the most challenging hurdles to overcome in developing novel treatments is the profound immune suppression found in many GBM patients. This limits the utility of all manner of immunotherapeutic agents, which have revolutionized the treatment of a number of cancers in recent years, but have failed to show similar benefit in GBM therapy. Understanding the mechanisms of tumor-mediated immune suppression in GBM is critical to the development of effective novel therapies, and reversal of this effect may prove key to effective immunotherapy for GBM. In this review, we discuss the current understanding of tumor-mediated immune suppression in GBM in both the local tumor microenvironment and systemically. We also discuss the effects of current GBM therapy on the immune system. We specifically explore some of the downstream effectors of tumor-driven immune suppression, particularly myeloid-derived suppressor cells (MDSCs) and other immunosuppressive monocytes, and the manner by which GBM induces their formation, with particular attention to the role of GBM-derived extracellular vesicles (EVs). Lastly, we briefly review the current state of immunotherapy for GBM and discuss additional hurdles to overcome identification and implementation of effective therapeutic strategies.


Electron spectroscopic imaging of encapsidated DNA in vaccinia virus.
We have used electron spectroscopic imaging to locate the phosphorus in vaccinia DNA in situ in unstained, ultrathin sections of virions. The phosphorus of the DNA backbone appeared to form a halo on the core periphery surrounding a phosphorus-impoverished central element. These results constrain models for how DNA could be packaged into mature vaccinia particles.


Microbial biofilms in biorefinery - Towards a sustainable production of low-value bulk chemicals and fuels.
Harnessing the potential of biocatalytic conversion of renewable biomass into value-added products is still hampered by unfavorable process economics. This has promoted the use of biofilms as an alternative to overcome the limitations of traditional planktonic systems. In this paper, the benefits and challenges of biofilm fermentations are reviewed with a focus on the production of low-value bulk chemicals and fuels from waste biomass. Our study demonstrates that biofilm fermentations can potentially improve productivities and product yields by increasing biomass retention and allowing for continuous operation at high dilution rates. Furthermore, we show that biofilms can tolerate hazardous environments, which improve the conversion of crude biomass under substrate and product inhibitory conditions. Additionally, we present examples for the improved conversion of pure and crude substrates into bulk chemicals by mixed microbial biofilms, which can benefit from microenvironments in biofilms for synergistic multi-species reactions, and improved resistance to contaminants. Finally, we suggest the use of mathematical models as useful tools to supplement experimental insights related to the effects of physico-chemical and biological phenomena on the process. Major challenges for biofilm fermentations arise from inconsistent fermentation performance, slow reactor start-up, biofilm carrier costs and carrier clogging, insufficient biofilm monitoring and process control, challenges in reactor sterilization and scale-up, and issues in recovering dilute products. The key to a successful commercialization of the technology is likely going to be an interdisciplinary approach. Crucial research areas might include genetic engineering combined with the development of specialized biofilm reactors, biofilm carrier development, in-situ biofilm monitoring, model-based process control, mixed microbial biofilm technology, development of suitable biofilm reactor scale-up criteria, and in-situ product recovery.


Fructose intolerance in children presenting with abdominal pain.
We determined the occurrence of fructose malabsorption in pediatric patients with previous diagnoses of abdominal pain caused by a functional bowel disorder, whether the restriction of fructose intake changes the reporting of symptoms, the role of fructose dosage, and the severity of resultant symptoms. We administered a fructose breath test to children presenting with persistent unexplained abdominal pain. Patients randomly received 1, 15, or 45 g fructose, and breath hydrogen was measured for 3 hours after ingestion. Test results were positive when breath hydrogen was 20 ppm greater than baseline and was accompanied by gastrointestinal symptoms. A total of 32 patients was enrolled, and none of the 9 who received 1 g had positive results. Three of 10 who received 15 g and 8 of 13 who received 45 g had positive results. All patients with positive test results restricted their fructose intake. Among the group with positive results, 9 of 11 had rapid improvement of their gastrointestinal symptoms. After 2 months, all 9 patients continued to report improvement. We concluded that fructose malabsorption may be a significant problem in children and that management of dietary intake can be effective in reducing gastrointestinal symptoms.


Comparative pharmacokinetics of four bioactive components in normal and chronic heart failure rats after oral administration of Qiangxin Lishui Prescription by microdialysis combined with ultra-high-performance liquid chromatography.
Qiangxin Lishui Prescription (QLP) has been clinically applied for treating heart failure with remarkable curative effects. A multi-component pharmacokinetic research is very necessary for determining active substances in it. This study aims to profile the traits and differences in the pharmacokinetics of salvianolic acid B, astragaloside IV, calycosin-7-O-β-D-glucoside and kaempferol in QLP between normal and chronic heart failure (CHF) rats by microdialysis combined with ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Sensitive, selective, and online microdialysis combined with the UHPLC-MS/MS method was successfully established and applied to study the pharmacokinetics of QLP. The pathological condition of CHF could lead to the enhancement of systematic exposure and reduction of the metabolic rate of four bioactive components for better bioavailability and therapeutic efficacy. The pharmacokinetic results will provide data support for the clinical application of QLP.


Is There Any Improvement In Efficacy Of Xtracorporeal Shockwave Lithotripsy Therapy For Treating Renal Stones With Invent Of New Shockwave Machines?
Renal stone disease is a very common medical problem in general population. As with invent of newer therapeutic modalities, ESWL is already losing its popularity. But we believe it as an effective way of treating renal stones. This study was conducted to evaluate any improvement in success rate of ESWL therapy for treating renal stones with latest shockwave lithotripsy machines. Study conducted from June 2016 until November 2017 in Alkhor hospital, Hamad Medical Corporation, Qatar. All patients undergoing ESWL for renal stones in mentioned period were included. Total 197 patients underwent ESWL using newer machines. Factors already studied to affect the success rate like stone size, location, consistency (measured by Hounsfield units on CT), presence of stent were taken in consideration. After a period of 3 months either complete stone clearance or stone fragments smaller than 4 mm were considered as a treatment success. These results were compared to the results from literature. Patients were followed until 3 months after treatment. 170 patients (86.29%) had complete stone clearance. Eleven patients (5.58%) had residual stone less than 4 mm, thus achieving an overall success of 181 patients (91.88%). 42 patients (21.32%) needed repeat session of ESWL with a maximum number of 3 sessions. 16 patient's (8.12%) required auxiliary procedures like flexible ureteroscopy. Post-ESWL complications were recorded in 12 patients (6.09%). Success rate was affected mainly by stone size with negligible effect of stone location. Presence of stent affected the number of sessions but has no impact on stone clearance.. Although the stone size and to a negligible extent, the stone location and presence of stent may affect the stone clearance, nevertheless a significant improvement in success rate has been observed by use of new shockwave lithotripsy machines.


Targeting mast cells tryptase in tumor microenvironment: a potential antiangiogenetic strategy.
Angiogenesis is a complex process finely regulated by the balance between angiogenesis stimulators and inhibitors. As a result of proangiogenic factors overexpression, it plays a crucial role in cancer development. Although initially mast cells (MCs) role has been defined in hypersensitivity reactions and in immunity, it has been discovered that MCs have a crucial interplay on the regulatory function between inflammatory and tumor cells through the release of classical proangiogenic factors (e.g., vascular endothelial growth factor) and nonclassical proangiogenic mediators granule-associated (mainly tryptase). In fact, in several animal and human malignancies, MCs density is highly correlated with tumor angiogenesis. In particular, tryptase, an agonist of the proteinase-activated receptor-2 (PAR-2), represents one of the most powerful angiogenic mediators released by human MCs after c-Kit receptor activation. This protease, acting on PAR-2 by its proteolytic activity, has angiogenic activity stimulating both human vascular endothelial and tumor cell proliferation in paracrine manner, helping tumor cell invasion and metastasis. Based on literature data it is shown that tryptase may represent a promising target in cancer treatment due to its proangiogenic activity. Here we focused on molecular mechanisms of three tryptase inhibitors (gabexate mesylate, nafamostat mesylate, and tranilast) in order to consider their prospective role in cancer therapy.


The treatment of malaria.
At least four doses of quinine followed by a single dose of mefloquine or by a single dose of sulfadoxine-pyrimethamine are two highly effective regimens for chloroquine-resistant falciparum malaria. Mefloquine alone is valuable in ambulant patients. Chloroquine-sensitive falciparum malaria can be treated with a course of chloroquine. Vivax and all other types of malaria should be treated with sequential chloroquine and primaquine. Quinine, by intravenous infusion, is the most effective drug for severe falciparum malaria. The optimum intravenous dose varies between 5 mg/kg and 10 mg/kg administered over four hours. Intravenous or oral quinine should be administered about every 12 hours and the total daily dose of quinine should rarely exceed 20 mg/kg. Intravenous fluid input should be controlled in falciparum malaria to prevent pulmonary oedema. Established renal failure is best treated by dialysis. The value of adrenocortical steroids for falciparum coma has not been established. Fresh blood transfusion may be helpful in small doses for severe anaemia and to replace clotting factors. Anticoagulants, such as heparin, should not be used in falciparum malaria.


The stability of fixation of proximal femoral fractures: a radiostereometric analysis.
The aim of this study was to quantify the stability of fracture-implant complex in fractures after fixation. A total of 15 patients with an undisplaced fracture of the femoral neck, treated with either a dynamic hip screw or three cannulated hip screws, and 16 patients with an AO31-A2 trochanteric fracture treated with a dynamic hip screw or a Gamma Nail, were included. Radiostereometric analysis was used at six weeks, four months and 12 months post-operatively to evaluate shortening and rotation. Migration could be assessed in ten patients with a fracture of the femoral neck and seven with a trochanteric fracture. By four months post-operatively, a mean shortening of 5.4 mm (-0.04 to 16.1) had occurred in the fracture of the femoral neck group and 5.0 mm (-0.13 to 12.9) in the trochanteric fracture group. A wide range of rotation occurred in both types of fracture. Right-sided trochanteric fractures seem more rotationally stable than left-sided fractures. This prospective study shows that migration at the fracture site occurs continuously during the first four post-operative months, after which stabilisation occurs. This information may allow the early recognition of patients at risk of failure of fixation.


Alcohol-induced changes of beta-lactoglobulin-retinol-binding stoichiometry.
It has been demonstrated using CD that ethanol induces important secondary structure changes of beta-lactoglobulin. CD spectra indicate that beta-lactoglobulin secondary structure, which is mainly composed of beta-strands, becomes mostly alpha-helical under the influence of the solvent polarity changes. The midpoint of beta-strand/alpha-helix transition in beta-lactoglobulin is observed at dielectric constant approximately 60 (35% ethanol; v/v). According to CD measurements, the ethanol-dependent secondary structure changes are reversible. The alkylation of lysines epsilon-NH2 in beta-lactoglobulin weakens the central beta-barrel structure, since the beta-strand/alpha-helix transition midpoint of alkylated beta-lactoglobulin is shifted to lower ethanol concentration (25% ethanol; v/v). beta-Lactoglobulin structural changes are triggering the dissociation of the beta-lactoglobulin-retinol complex as judged from complete quenching of its fluorescence in ethanol concentration greater than 30% (v/v). However, in 20% ethanol (v/v), beta-lactoglobulin still retains most of its native secondary structure as shown by CD and, in this condition, one beta-lactoglobulin molecule binds an additional second retinol molecule. This suggests that the highly populated species observed around 20% ethanol (v/v) might represent an intermediate state able to bind two molecules of retinol.


Successful Non-Operative Management of Adhesive Small Bowel Obstruction: Is it Really a Success?
In adhesive small bowel obstructions (ASBOs), literature has shown that passage of a water-soluble contrast challenge at either 8 hours or 24 hours is predictive of successful non-operative management (NOM) for an ASBO, but the long-term outcomes between these two groups are unknown. We hypothesized that patients who require longer transit times to the colon have a higher one-year recidivism of ASBO. This was a 4-year review of patients with presumed ASBO undergoing successful NOM. Those requiring operation or those with an SBO due to something other than adhesions were excluded. Patients were divided into two groups (8 hour and 24 hour) based on when contrast reached their right colon. Patients were followed for one year to determine ASBO recurrence. 137 patients underwent NOM; 112 in the 8-hour group and 25 in the 24-hour group. One-year recurrence rate was 21.4% in the 8-hour group and 40% in the 24-hour group (P = 0.05). The median time to recurrence was 113 days in the 8-hour group and 13 days in the 24-hour group (P = 0.02). Of those who recurred in the 24-hour group, 60% recurred within 30 days (P = 0.01). On univariable analysis, first-time ASBO and 24-hour transit time were risk factors for recurrence. Adhesive small bowel obstruction patients undergoing NOM in the 24-hour group had a recurrence rate nearly twice that of patients in the 8-hour group and may benefit from an operative exploration during the index hospitalization at the 8-hour mark of a water-soluble contrast challenge, especially if experiencing a first-time ASBO.


Listeria monocytogenes F2365 carries several authentic mutations potentially leading to truncated gene products, including inlB, and demonstrates atypical phenotypic characteristics.
Searches of the genome annotation of Listeria monocytogenes F2365, an isolate from the 1985 listeriosis epidemic in California, showed that this strain carries 20 authentic mutations resulting in premature stop codons, including a nonsense mutation in inlB. Here we showed that L. monocytogenes F2365 demonstrates atypical virulence-associated characteristics, including significantly (P < 0.05) reduced invasion efficiency in Caco-2 cells as compared with a closely related lineage I serotype 4b strain as well as significantly (P < 0.05) greater variation in invasiveness when grown under different conditions compared with standard laboratory control and other lineage I serotype 4b strains. In addition, L. monocytogenes F2365 demonstrated distinct growth characteristics, including a significantly (P < 0.05) reduced exponential growth rate when compared with laboratory control and other lineage I serotype 4b outbreak-associated strains as well as a significantly (P < 0.05) longer lag phase duration time compared with another lineage I serotype 4b strain. Our results support that L. monocytogenes F2365 is characterized by genotypic and phenotypic properties that are atypical of other L. monocytogenes strains.


Isolation of the membrane-bound 26 000-Mr penicillin-binding protein of Streptomyces strain K15 in the form of a penicillin-sensitive D-alanyl-D-alanine-cleaving transpeptidase.
The membrane-bound, 26 000-Mr penicillin-binding protein of Streptomyces K15 has been isolated in the form of an effective, penicillin-sensitive D-alanyl-D-alanine-cleaving peptidase exhibiting high transpeptidase activity (greater than 95%) and very low carboxy-peptidase activity (less than 5%). The penicillin-binding protein/transpeptidase can be extracted directly from the mycelium with N-cetyl-NNN-trimethylammonium bromide (Cetavlon) and subsequently obtained at 90% purity and with an 8000-fold specific enrichment (when compared with the activity of the isolated membranes) by a two-step procedure involving Sephadex filtration and affinity chromatography on ampicillin-linked CH Sepharose 4B in the presence of detergent. At saturating concentrations of the co-substrates diacetyl-L-Lys-D-Ala-D-Ala and Gly-Gly, the catalytic-centre activity is about 0.3 s-1.


The impact of meningococcal polymerase chain reaction testing on laboratory confirmation of invasive meningococcal disease.
Laboratory methods of diagnosis were examined for 266 children with invasive meningococcal disease. Seventy-five (36%) of 207 cases with bloodstream infection had both positive blood culture and blood meningococcal polymerase chain reaction (PCR), 130 (63%) negative blood culture and positive blood PCR, and 2 (1%) had positive blood culture and negative blood PCR. Sixty-three percent of cases were diagnosed by PCR alone.


Intravitreal injection of methotrexate in an experimental rabbit model: determination of pharmacokinetics.
To investigate the pharmacokinetics of intravitreally administered methotrexate. Twenty-one New Zealand white rabbits were used in the study. The pharmacokinetics of intravitreally injected 800 μg/0.1 ml of methotrexate was investigated. Intravitreal concentration of the drug was measured at seven different times, in six eyes at each occasion, on a total of 42 eyes of 21 rabbits from a period of 30 minutes to 72 hours. The volume of distribution was calculated as 1.33 ml following intravitreal injection of 800 μg methotrexate. Vitreous concentrations of the drug were found to be decreasing related to the specific mathematical equation; drug concentration= 1426.73 e -0.1182(time) and remained over effective dose by 81 hours with a half life of 5.9 hours. These findings evidenced those vitreous levels of methotrexate at various time intervals after 800 μg intravitreal injections which formulated a mathematical equation for calculation of vitreous level of the drug at each hour.


Nanoparticle heterogeneity: an emerging structural parameter influencing particle fate in biological media?
Drug nanocarriers' surface chemistry is often presumed to be uniform. For instance, the polymer surface coverage and distribution of ligands on nanoparticles are described with averaged values obtained from quantification techniques based on particle populations. However, these averaged values may conceal heterogeneities at different levels, either because of the presence of particle sub-populations or because of surface inhomogeneities, such as patchy surfaces on individual particles. The characterization and quantification of chemical surface heterogeneities are tedious tasks, which are rather limited by the currently available instruments and research protocols. However, heterogeneities may contribute to some non-linear effects observed during the nanoformulation optimization process, cause problems related to nanocarrier production scale-up and correlate with unexpected biological outcomes. On the other hand, heterogeneities, while usually unintended and detrimental to nanocarrier performance, may, in some cases, be sought as adjustable properties that provide NPs with unique functionality. In this review, results and processes related to this issue are compiled, and perspectives and possible analytical developments are discussed.


Metformin Attenuates Ischemia-reperfusion Injury of Fatty Liver in Rats Through Inhibition of the TLR4/NF-κB Axis
Donor organs for liver transplantation may often have fatty liver disease, which confers a higher sensitivity to ischemia/reperfusion injury. At present, there is no effective treatment for the condition. Evidence has suggested that metformin, the first-line medication for diabetes, has protective effects against many disorders. However, the potential role of metformin in ischemia/reperfusion injury in fatty liver disease remains unclear. To examine the effect of metformin treatment during ischemia/reperfusion injury in fatty liver and determine the possible mechanisms. Animal experimentation. Sprague-Dawley male rats were fed a high-fat diet (520 kcal/100 g) for 14 weeks and then were subjected to the orthotopic autologous liver transplantation model. Sections of liver tissue were stained with hematoxylin and eosin to visualize the damage. Blood and liver samples were used to analyze the related proteins and components involved in the inflammatory signaling pathway. We found that metformin significantly ameliorated the ischemia/reperfusion injury of the fatty liver through a reduction in alanine aminotransferase/aspartate aminotransferase concentrations in the serum and a decrease in dead cells, as shown by the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay (p<0.05). In addition, metformin significantly attenuated interleukin (IL)-6, IL-1β, and tumor necrosis factor-α production and increased the expression of active caspase-3 and Bax in the liver (p<0.05). Mechanistically, metformin suppressed the activation of toll-like receptor 4 (TLR4)/NF-κB signaling (p<0.05), resulting in a decreased inflammatory response and apoptosis. Our findings demonstrated that metformin attenuated ischemia/reperfusion injury in fatty liver disease via the TLR4/NF-κB axis, suggesting that metformin could have potential therapeutic applications in ischemia/reperfusion injury associated with liver transplantation.


A kinesin with calponin-homology domain is involved in premitotic nuclear migration.
Interaction and cross-talk between microtubules and actin microfilaments are important for numerous processes during plant growth and development, including the control of cell elongation and tissue expansion, but little is known about the molecular components of this interaction. Plant kinesins with the calponin-homology domain (KCH) were recently identified and associated with a putative role in microtubule-microfilament cross-linking. The putative biological role of the rice KCH member OsKCH1 is addressed here using a combined approach with Tos17 kch1 knock-out mutants on the one hand, and a KCH1 overexpression line generated in tobacco BY-2 cells. It is shown that OsKCH1 is expressed in a development and tissue-specific manner in rice and antagonistic cell elongation and division phenotypes as a result of knock-down and overexpression are reported. Further, the dynamic repartitioning of OsKCH1 during the cell cycle is described and it is demonstrated that KCH overexpression delays nuclear positioning and mitosis in BY-2 cells. These findings are discussed with respect to a putative role of KCHs as linkers between actin filaments and microtubules during nuclear positioning.


Parents' evaluation of developmental status: how well do parents' concerns identify children with behavioral and emotional problems?
This study was undertaken to determine which parental concerns are most associated with significant behavioral/emotional problems and the extent to which parents' concerns can be depended on in the detection of mental health problems. An additional goal is to view how well a recently published screening test relying on parents' concerns, Parents' Evaluation of Developmental Status (PEDS), detects behavioral and emotional problems. Subjects were a national sample of 472 parents and their children (21 months to 8 years old) who were participants in 1 of 2 test standardization and validation studies. Sites included various pediatric settings, public schools, and Head Start programs in 5 diverse geographic locations. Subjects were representative of U.S. demographics in terms of ethnicity, parental level of education, gender, and socioeconomic status. At each site, psychological examiners, educational diagnosticians, or school psychologists recruited families, and obtained informed consent. Examiners disseminated a demographics questionnaire (in English or Spanish) and a developmental screening test that relies on parents' concerns (PEDS). Examiners were blinded to PEDS' scoring and interpretation administered either by interview or in writing, the Eyberg Child Behavior Inventory (ECBI) or the Possible Problems Checklist (PPC), a subtest of the Child Development Inventory that includes items measuring emotional well-being and behavioral self-control. PEDS was used to sort children into risk for developmental disabilities according to various types of parental concern. Those identified as having high or moderate risk were nominated for diagnostic testing or screening followed by developmental and mental health services when indicated. Because their emotional and behavioral needs would have been identified and addressed, these groups were removed from the analysis (N = 177). Of the 295 children who would not have been nominated for further scrutiny on PEDS due to their low risk of developmental problems, 102 had parents with concerns not predictive of developmental disabilities (e.g., behavior, social skills, self-help skills) and 193 had no concerns at all. Of the 295 children, 12% had scores on either the ECBI or the PPC indicative of mental health problems. Two parental concerns were identified through logistic regression as predictive of mental health status: behavior (OR = 4.74, CI = 1.69-13.30); and social skills (OR = 5.76, CI = 2.46-13.50). If one or more of these concerns was present, children had 8.5 times the risk of mental health problems (CI = 3.69-19.71) In children 434 years of age and older, one or both concerns was 87% sensitive and 79% specific to mental health status, figures keeping with standards for screening test accuracy. In young children, the presence of one or both concerns was 68% sensitive and 66% specific to mental health status. The findings suggest that certain parental concerns, if carefully elicited, can be depended on to detect mental health problems when children are 41 years and older and at low risk of developmental problems. For younger children, clinicians should counsel parents in disciplinary techniques, follow up, and if suggestions were not effective, administer a behavioral-emotional screening test such as the Pediatric Symptoms Checklist or the ECBI before making a referral decision.


Artificial intelligence and cognitive diagnosis based teaching resource recommendation algorithm.
In the realm of advanced technology, deep learning capabilities are harnessed to analyze and predict novel data, once it has absorbed existing information. When applied to the sphere of education, this transformative technology becomes a catalyst for innovation and reform, leading to advancements in teaching modes, methodologies, and curricula. In light of these possibilities, the application of deep learning technology to teaching resource recommendations is explored in this article. Within the context of the study, a bespoke recommendation algorithm for teaching resources is devised, drawing upon the integration of deep learning and cognitive diagnosis (ADCF). This intricately constructed model consists of two core elements: the Multi-layer Perceptron (MLP) and the Generalized Matrix Factorization (GMF), operating cohesively through stages of linear representation and nonlinear learning of the interaction function. The empirical analysis reveals that the ADCF model achieves 0.626 and 0.339 in the hits ratio (HR) and the Normalized Discounted Cumulative Gain (NDCG) respectively due to the traditional model, signifying its potential to add significant value to the domain of teaching resource recommendations.


Respiratory indices during sleep in healthy infants: A prospective longitudinal study and meta-analysis.
Healthy infants may have a greater apnea hypopnea index (AHI) than older children during the newborn period, but the trajectory of these sleep-related events beyond the first month of life is poorly understood. In this study, we evaluated the longitudinal changes in respiratory indices during sleep in healthy infants during the first six months of life. Single-center prospective cohort study. Thirty healthy infants underwent overnight in-lab polysomnography at one and five months of age and findings were compared between assessments. Systematic review of studies evaluating infant polysomnography and meta-analysis was conducted. At one month of age, total AHI, obstructive AHI, and central AHI model-adjusted means (95% confidence interval) were 16.9 events/hour (12.2, 21.5), 10.2 events/hour (7.4, 13.1), and 6.6 events/hour (4.2, 9.0), respectively. 16.8% of events were obstructive apneas and 36.1% central apneas. By five months of age, there were significant reductions in each index to 4.1 events/hour (3.2, 5.0), 1.9 events/hour (1.4, 2.4), and 2.2 events/hour (1.6, 2.9), respectively (p < 0.001 for each), and a lower proportion of events were obstructive apneas (8.6%, p = 0.007) and a greater proportion central apneas (52.3%, p = 0.002). Meta-analysis found high AHI in infants with significant heterogeneity. Central AHI and obstructive AHI are greater in healthy newborns than older children. There is a significant spontaneous reduction in events and change in type of events in the first six months of life in this low-risk population. These findings may serve as a reference for clinicians evaluating for obstructive sleep apnea in infants.


Analysis of antibody response to invariable regions of VlsE, the variable surface antigen of Borrelia burgdorferi.
VlsE, the variable surface antigen of Borrelia burgdorferi, consists of two invariable domains at the amino and carboxyl termini and one central variable domain. The latter contains six invariable regions, IR(1) to IR(6), and six variable regions. In the present study, the antigenicity of all of the invariable regions in B. burgdorferi-infected monkeys, humans, and mice was assessed by peptide-based enzyme-linked immunosorbent assays. Only one invariable region, IR(6), was antigenic in all animals of the three host species. IR(2) and IR(4) were also antigenic in mice.


Favorable progression-free survival in women with two different histopathological subtypes of bilateral breast cancer.
The aim of this study was to classify bilateral breast cancers (BBCs) into two groups according to histopathological or molecular subtypes of the two breast diseases in each patient and to study their characteristics in relation to survival outcomes. Fifty-six BBC patients were enrolled in the study. They were classified according to whether the two breast diseases were of the same or different histopathological subtypes (defined as S-his or D-his groups) and molecular subtypes (defined as S-mole and D-mole groups). Progression-free survival (PFS), overall survival (OS), and important characteristics were then compared between the groups. We observed that the PFS and OS of the S-mole and D-mole groups failed to reach a significant difference. However, D-his BBC patients enjoyed longer PFS than their S-his counterparts, although the OS was similar. To explore the reason for the extended PFS in D-his BBC patients, we first focused on the possible prognostic contribution by various histopathological subtypes in the groups. We compared the proportion of infiltrating ductal carcinoma, infiltrating lobular carcinoma, and other breast cancer subtypes in the D-his and S-his groups, and demonstrated that they were not associated with longer PFS. We then examined age, menopausal status, tumor, node, and metastasis (TNM) stage, expressions of estrogen receptor (ER), progesterone receptor (PR) and Ki67, and metastasis to lymph nodes, viscera, and bone. The results indicated no significant between-group differences in age, TNM stage, ER/PR expression, and metastasis to viscera and bone. However, significantly lower levels of Ki67and decreased lymph node metastasis rate were associated with D-his BBC patients, which might explain the observed longer PFS. In comparison to S-his, D-his BBC patients had longer PFS, which was associated with lower levels of Ki67 and a decreased lymph node metastasis rate.


Mechanisms of physiological adjustment of N2 fixation in Cicer arietinum L. (chickpea) during early stages of water deficit: single or multi-factor controls.
Drought negatively impacts symbiotic nitrogen fixation (SNF) in Cicer arietinum L. (chickpea), thereby limiting yield potential. Understanding how drought affects chickpea nodulation will enable the development of strategies to biotechnologically engineer chickpea varieties with enhanced SNF under drought conditions. By analyzing carbon and nitrogen metabolism, we studied the mechanisms of physiological adjustment of nitrogen fixation in chickpea plants nodulated with Mesorhizobium ciceri during both drought stress and subsequent recovery. The nitrogenase activity, levels of several key carbon (in nodules) and nitrogen (in both nodules and leaves) metabolites and antioxidant compounds, as well as the activity of related nodule enzymes were examined in M. ciceri-inoculated chickpea plants under early drought stress and subsequent recovery. Results indicated that drought reduced nitrogenase activity, and that this was associated with a reduced expression of the nifK gene. Furthermore, drought stress promoted an accumulation of amino acids, mainly asparagine in nodules (but not in leaves), and caused a cell redox imbalance in nodules. An accumulation of organic acids, especially malate, in nodules, which coincided with the decline of nodulated root respiration, was also observed under drought stress. Taken together, our findings indicate that reduced nitrogenase activity occurring at early stages of drought stress involves, at least, the inhibition of respiration, nitrogen accumulation and an imbalance in cell redox status in nodules. The results of this study demonstrate the potential that the genetic engineering-based improvement of SNF efficiency could be applied to reduce the impact of drought on the productivity of chickpea, and perhaps other legume crops.


Aberrant transcriptional regulatory network in T cells of multiple sclerosis.
To identify the molecular network of the genes deregulated in multiple sclerosis (MS), we studied gene expression profile of purified CD3(+) T cells isolated from Hungarian monozygotic MS twins by DNA microarray analysis. By comparing three concordant and one discordant pairs, we identified 20 differentially expressed genes (DEG) between the MS patient and the genetically identical healthy subject. Molecular network of 20 DEG analyzed by KeyMolnet, a comprehensive information platform, indicated the close relationship with transcriptional regulation by the Ets transcription factor family and the nuclear factor NF-kappaB. This novel bioinformatic approach proposes the logical hypothesis that aberrant regulation of the complex transcriptional regulatory network contributes to development of pathogenic T cells in MS.


Characterization and classification of external quality assessment schemes (EQA) according to objectives such as evaluation of method and participant bias and standard deviation. External Quality Assessment (EQA) Working Group A on Analytical Goals in Laboratory Medicine.
Within the scope of this paper, the Working Group has attempted to place external quality assessment (EQA) within the whole context of quality management in laboratory medicine. First, the objectives of EQA schemes are defined and current EQA schemes evaluated. In most schemes, the objectives are not defined a priori and do not allow the definition of the origin of unacceptable individual results from participants. There is an ongoing trend for making traditional EQA schemes more interesting for the participants. Analysis of the factors involved in analytical quality allow the definition of the essential analytical tasks of educational EQA schemes. Beside these quality control tasks, educational EQA also includes quality assurance elements. EQA today has not only an important role to play in the assessment of each participant's performance but also in the assessment of the method. Efficiency of the schemes and educational impact can be improved by appropriate scheme designs according to objectives. After this theoretical approach, some practical examples of problem related EQA designs are given.