Nevus size and number are associated with telomere length and represent potential markers of a decreased senescence in vivo.
Nevus counts represent one of the strongest risk factors for melanoma. They appear in childhood and adolescence and involute from middle age onwards. Recent evidence has shown that nevus cells undergo oncogene-induced senescence involving the p16/retinoblastoma pathway. However, telomere length also influences senescence in proliferative somatic cells and varies between individuals. This study explores whether telomere length measured in white cells is associated with nevus count and size in 1,897 Caucasian women ages 18 to 79 years. Total body nevus counts were positively correlated with white cell telomere length (mean, 7.09 kbp; range, 5.09-9.37) after adjustment for age (P = 0.0001). Age-adjusted telomere length was also associated with nevus count for nevi above 5 mm in diameter (P = 0.04). Subjects in the top category for nevus count had an average age-adjusted telomere length 150 bp longer than those in the lowest category. The positive correlation between white cell telomere length and nevi number and size may reflect an increased replicative potential (reduced senescence) in individuals with longer telomeres, which may not be melanocyte specific. Understanding mechanisms influencing the induction and involution of nevi will not only help in understanding the pathophysiology of melanoma but should also shed light on the complex relationship between aging and cancer.


Angiotensin II induces cellular hypertrophy in cultured murine proximal tubular cells.
In the present study we demonstrate that a murine proximal tubular cell line (MCT cells), expressing angiotensin II (ANG II) receptors [dissociation constant (Kd) = 0.89 nM; receptor density (R0) = 46,900 receptors/cell] in culture, can be induced to hypertrophy after the daily addition of exogenous ANG II (10(-8) M). This hypertrophic response was characterized by an increase in total cellular protein content, by an enhancement of [3H]leucine incorporation into precipitable proteins, and by an augmentation in cell size by cytofluorography. This ANG II effect producing MCT cell enlargement was demonstrable in the absence of cellular proliferation. Proliferation of MCT cells, however, could be induced by epidermal growth factor (EGF) or platelet-derived growth factor (PDGF), and pretreatment of rested MCT cells with ANG II further enhanced EGF-induced cell division. ANG II-induced hypertrophy in MCT cells was factor specific, in that it could be blocked with saralasin, and not induced by angiotensin I (ANG I). This hypertrophic response was also independent of prostaglandin E2 synthesis but was transducible by pertussis toxin-sensitive G proteins and involved, to some extent, the activation of Na(+)-H+ exchange. ANG II, as well as EGF and/or PDGF, moreover, could induce the cellular oncogenes c-fos, c-myc, c-N-ras, but not c-cis, which suggests that early gene activation is probably not a specific prerequisite for hypertrophy. Our findings demonstrate that ANG II, in culture, can be a single-factor event capable of inducing hypertrophy in proximal tubular cells.


Prescribing PPIs.
Proton pump inhibitors (PPIs) are widely prescribed and many people remain on them for years. In some, there may be good justification for long-term use; for example, in those with oesophageal stricture, Barrett's oesophagus or a history of a bleeding gastrointestinal ulcer, or to provide gastroprotection in those at high risk of gastrointestinal complications from taking NSAIDs. However, PPIs are often not used in line with clinical guidelines.1,2 In addition, there are concerns that many people take them for long periods to manage less serious conditions (e.g. indigestion) and may prefer taking them to addressing factors such as diet, obesity or alcohol that may be contributing to their symptoms. Although PPIs are well-tolerated, there is increasing evidence that they may be associated with a range of long-term adverse effects. Here, we review the safety of PPIs and consider whether long-term prescribing needs to be reassessed.


Warming Amplifies the Frequency of Harmful Algal Blooms with Eutrophication in Chinese Coastal Waters.
Widespread coastal eutrophication is known to increase the prevalence of harmful algal blooms (HABs). Increased HABs have also been linked to climate change, with ocean warming predicted to lead to increased prevalence and earlier timing of HABs. Testing the predictions of warming to HABs is difficult due to the lack of long-term observations across spatial scales. Here, we use a 45 year (1970-2015) record of the occurrence and duration of HABs along Chinese coast to show that the HAB frequency has increased at a rate of 40 ± 4% decade-1, with earlier timing by 5.50 ± 1.78 days decade-1. The increasing frequency of blooms varied with latitude and is significantly correlated with warming at an average rate of 0.17 ± 0.03 °C decade-1, with the positive relationship being strongest in more eutrophic provinces. HAB frequency increased with elevated dissolved inorganic nutrient concentration, but this increase was amplified further with warming. Warming and eutrophication showed additive roles in triggering HABs. Swift action to mitigate eutrophication is essential to avoid a sharp increase in the HABs in coastal waters with further warming.


Optical Control of the Topology of Laser-Plasma Accelerators.
We propose a twisted plasma accelerator capable of generating relativistic electron vortex beams with helical current profiles. The angular momentum of these vortex bunches is quantized, dominates their transverse motion, and results in spiraling particle trajectories around the twisted wakefield. We focus on a laser wakefield acceleration scenario, driven by a laser beam with a helical spatiotemporal intensity profile, also known as a light spring. We find that these light springs can rotate as they excite the twisted plasma wakefield, providing a new mechanism to control the twisted wakefield phase velocity and enhance energy gain and trapping efficiency beyond planar wakefields.


Identification of a transcription factor, PunR, that regulates the purine and purine nucleoside transporter punC in E. coli.
Many genes in bacterial genomes are of unknown function, often referred to as y-genes. Recently, the analytic methods have divided bacterial transcriptomes into independently modulated sets of genes (iModulons). Functionally annotated iModulons that contain y-genes lead to testable hypotheses to elucidate y-gene function. The inversely correlated expression of a putative transporter gene, ydhC, relative to purine biosynthetic genes, has led to the hypothesis that it encodes a purine-related transporter and revealed a LysR-family regulator, YdhB, with a predicted 23-bp palindromic binding motif. RNA-Seq analysis of a ydhB knockout mutant confirmed the YdhB-dependent activation of ydhC in the presence of adenosine. The deletion of either the ydhC or the ydhB gene led to a substantially decreased growth rate for E. coli in minimal medium with adenosine, inosine, or guanosine as the nitrogen source. Taken together, we provide clear evidence that YdhB activates the expression of the ydhC gene that encodes a purine transporter in E. coli. We propose that the genes ydhB and ydhC be re-named as punR and punC, respectively.


Tuning the potency and selectivity of ImmTAC molecules by affinity modulation.
T cell engaging bispecifics have great clinical potential for the treatment of cancer and infectious diseases. The binding affinity and kinetics of a bispecific molecule for both target and T cell CD3 have substantial effects on potency and specificity, but the rules governing these relationships are not fully understood. Using ImmTAC (Immune mobilizing monoclonal TCRs Against Cancer) molecules as a model, we explored the impact of altering affinity for target and CD3 on the potency and specificity of the re-directed T cell response. This class of bispecifics, exemplified by tebentafusp which has recently shown survival benefit in a randomized phase 3 clinical trial1, bind specific target peptides presented by human leukocyte antigen (HLA) on the cell surface via an affinity-enhanced T cell receptor and can redirect T cell activation with an anti-CD3 effector moiety. The data reveal that combining a strong affinity TCR with an intermediate affinity anti-CD3 results in optimal T cell activation, while strong affinity of both targeting and effector domains significantly reduces maximum cytokine release. Moreover, by optimising the affinity of both parts of the molecule, it is possible to improve the selectivity. These results could be effectively modelled based on kinetic proof-reading with limited signalling. This model explained the experimental observation that strong binding at both ends of the molecules leads to reduced activity, through very stable target-bispecific-effector complexes leading to CD3 entering a non-signalling dark-state. These findings have important implications for the design of anti-CD3 based bispecifics with optimal biophysical parameters for both activity and specificity.


Complications in the treatment of acute spinal injury.
Complications may occur during any phase of the treatment process for acute spinal injuries. An awareness of the most common problems is the most powerful tool that the physician has to prevent him or her from repeating at least some of the errors of others.


A Loop-mediated Isothermal Amplification With a Nanoparticle-Based Lateral Flow Biosensor Assay to Detect Pseudomonas aeruginosa in Endophthalmitis.
Pseudomonas aeruginosa is the most common bacteria causing endophthalmitis after cataract surgery. Vitreous fluid culture and molecular studies are commonly used in clinical diagnoses, but have disadvantages, such as a long culture cycle and low detection sensitivity. Here, we report a loop-mediated isothermal amplification (LAMP) method combined with the nanoparticles-lateral flow biosensor (LFB) method for rapid and specific detection of P. aeruginosa. A set of six primers was designed to target the OprL gene of P. aeruginosa. Genomic DNA extracted from several gram-negative and gram-positive bacteria was used to determine the sensitivity and specificity of the analysis. LAMP reactions were conducted at 65 °C for 50 minutes, and results were reported using the LFB method. The DNA template of P. aeruginosa was specifically recognized by the P. aeruginosa-LAMP-LFB (PA-LAMP-LFB) method as no cross reactions were observed for non-P. aeruginosa templates. The analytical sensitivity of our assay was 100 fg per test for the pure cultured DNA template, and the result obtained using the LFB was consistent with that of colorimetric indicator detection. The whole test could be completed within 1h. This method was used to detect P. aeruginosa, Staphylococcus aureus, and Klebsiella pneumoniae; only P. aeruginosa was positive. The positive rates of P. aeruginosa detected by a traditional culture method, the LAMP-LFB method, and the fluorescence quantitative polymerase chain reaction method were 17.7%, 17.7%, and 13.3%, respectively. The P. aeruginosa-LAMP-LFB method established here is a rapid, specific, and sensitive method for the detection of P. aeruginosa, which can be widely used.


Vesicular stomatitis virus-mediated cell fusion subsequent to virus adsorption at different pH values.
Vesicular stomatitis virus (VSV)-mediated cell fusion from without can be induced by transient exposure to low pH, subsequent to adsorption of VSV at neutral pH. To study the mechanism of VSV-induced cell fusion, we examined the effect of pH condition at virus adsorption on acid-inducible VSV-mediated cell fusion. Although the binding of VSV to BHK-21 cells was most efficient under acidic condition (pH 5.7-6.3), extensive cell fusion was not observed under this condition. A temporary exposure to low pH after binding at neutral pH also decreased fusion activity. However, return to neutral pH for 2 min just after the acid binding restored the fusion activity. These results indicate the requirement of neutral pH condition for VSV-mediated cell fusion prior to the acid stimulation which induces conformational change of the virus glycoprotein into a fusogenic form.


Laparoscopic versus vaginal hysterectomy for benign pathology.
The following review is an attempt to clarify the current role of vaginal and laparoscopic approaches in case of hysterectomy for benign pathologies. Recent researches establish that vaginal hysterectomy seems to be the gold standard in case of benign pathologies and should be performed in preference to abdominal hysterectomy wherever possible. When vaginal hysterectomy is not technically possible, laparoscopic hysterectomy is to be preferred to abdominal hysterectomy. No advantages of laparoscopic hysterectomy could be found over vaginal hysterectomy, in particular, because laparoscopic hysterectomy is associated with a higher rate of complications (especially bladder and ureteral injuries). Other authors, instead, show that laparoscopic hysterectomy permits a safe bilateral salpingo-oophorectomy (BSO), the treatment of additional pathologies at the time of surgery, a reduction of intraoperative bleeding, postoperative pelvic pain and length of stay compared with vaginal hysterectomy. Moreover, laparoscopic approach, in experienced hands, is not associated with any increase in major complication rate. It's time to reduce abdominal hysterectomy in favour of mini-invasive approaches. To do so a suitable training and supervision are paramount before embarking on total laparoscopic hysterectomy (so that complications are minimized) or on vaginal hysterectomy (so to perform a planned BSO). It is also essential, particularly for total laparoscopic hysterectomy, to share the techniques used by different surgeons, the results and the complications concerning this approach.


Extracellular HIV-1 Tat protein induces a rapid and selective activation of protein kinase C (PKC)-alpha, and -epsilon and -zeta isoforms in PC12 cells.
The addition in culture of extracellular HIV-1 Tat protein (0.1-1 nM) to PC12 cells induced a rapid increase of the bulk protein kinase C (PKC) catalytic activity. Among various PKC isoforms (alpha, beta I, beta II, delta, epsilon, eta, theta, and zeta) expressed in PC12 cells, Tat selectively stimulated alpha, epsilon, and zeta, as judged by activities in immunoprecipitates. Activation of these isoforms was suppressed by the tyrosine kinase inhibitor genistein. Moreover, PKC-zeta showed the fastest kinetics of activation in response to Tat, but PKC-alpha and PKC-epsilon showed the highest levels of activation. PKC-alpha activation was accompanied by a rise of intracellular IP3, while the PI 3-kinase inhibitors wortmannin and LY294002 suppressed PKC-epsilon activation. Taken together, these findings demonstrate that extracellular Tat shows a cytokine-like activity in PC12 cells, being able to trigger an intracellular signalling cascade which involves PKC-alpha, -epsilon, and -zeta.


[Formalin fixation in the method of the Ixodes persulcatus (ixodidae) age estimation by the state of fat reserves].
Formalin fixation is proved to be the useful tool in the method of age estimation for the taiga ticks collected in field. Formalin does not deform fixed material and does not dissolve fat; hence, it is the best fixing fluid when tick age is estimated by the state of fat reserves in its organism. The ticks preserved in 9% formalin were shown to be suitable for age estimation 1.5-2 months after fixation.


The clinical importance of fungal biofilms.
Fungal biofilms have become an increasingly important clinical problem. The widespread use of antibiotics, frequent use of indwelling medical devices, and a trend toward increased patient immunosuppression have resulted in a creation of opportunity for clinically important yeasts and molds to form biofilms. This review will discuss the diversity and importance of fungal biofilms in the context of clinical medicine, provide novel insights into the clinical management of fungal biofilm infection, present evidence why these structures are recalcitrant to antifungal therapy, and discuss how our knowledge and understanding may lead to novel therapeutic intervention.


Rod and cone photoreceptor cells produce ROS in response to stress in a live retinal explant system.
The production of reactive oxygen species (ROS) can lead to oxidative stress, which is a strong contributory factor to many ocular diseases. In this study, the removal of trophic factors is used as a model system to investigate the effects of stress in the retina. The aims were to determine if both rod and cone photoreceptor cells produce ROS when they are deprived of trophic factor support and to demonstrate if the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (Nox) enzymes are responsible for this ROS production. Retinas were explanted from mice aged between postnatal days 8-10 and cultured overnight. The following morning, confocal microscopy combined with various fluorescent probes was used to detect the production of ROS. Each time peanut agglutinin (PNA), a cone photoreceptor marker, was used to facilitate orientation of the retina. Dihydroethidium and dihydrorhodamine 123 (DHR123) were used to determine which cells produce ROS. Subsequently, western blots of retinal serial sections were used to detect the presence of Noxs in the different retinal layers. The Nox inhibitor apocynin was then tested to determine if it altered the production of ROS within these cells. Live retinal explants, viewed at high magnifications using confocal microscopy, displayed an increase in the fluorescent products of dihydroethidium and DHR123 upon serum removal when compared to controls. DHR123 fluorescence, once oxidized, localized to mitochondria and was found in the same focal plane as the PNA staining. This showed that cones and rods produced ROS when stressed. Retinal serial sectioning established that the photoreceptor layer expressed Nox4, dual oxidase (Duox) 1, and Duox2 at varying levels. Finally, the Nox inhibitor apocynin decreased the burst stimulated by the stress of serum removal. Confocal microscopy and PNA staining allowed differentiation of cell types within the outermost layers of the retina, demonstrating that both rods and cones generated ROS in response to the stress of serum deprivation. Nox4 was the most abundantly expressed Nox in the photoreceptor layer, but Duox1 and Duox2 were also present at detectable levels, and as apocynin reduced the levels of ROS produced, this implied that these proteins may play some role in this production.


AMF Inoculum Enhances Crop Yields of Zea mays L. 'Chenghai No. 618' and Glycine max L. 'Zhonghuang No. 17' without Disturbing Native Fugal Communities in Coal Mine Dump.
For the agricultural development of dumps, increase in land use efficiency and protection of food security, to verify the safety, efficacy and sustainability of field-applied arbuscular mycorrhizal fungi (AMF) inoculum, and to exclude the risk of potential biological invasion, in this study, we determined the effect of AMF inoculation and intercropping patterns (maize-soybean) on the temporal dynamics of soil parameters, native AMF communities and crop yields. AMF communities were analyzed using Illumina MiSeq. A total of 448 AMF operational taxonomic units (OTUs) belonging to six genera and nine families were identified. AMF inoculation treatment significantly improved the yield of intercropping maize and increased the content of available phosphorus. AMF diversity was significantly influenced by cropping pattern and growth stage, but not by the inoculation treatment. Inoculation altered the AMF community composition in the early growth stage and facilitated a more complex AMF network in the early and late growth stages. These results indicate that AMF inoculation affects native AMF only in the early stage, and its impact on yield may be the consequence of cumulative effects due to the advantages of plant growth and nutrient uptake in the early stage.


Ability of nucleoside-modified mRNA to encode HIV-1 envelope trimer nanoparticles.
The success of nucleoside-modified mRNAs in lipid nanoparticles (mRNA-LNP) as COVID-19 vaccines heralded a new era of vaccine development. For HIV-1, multivalent envelope (Env) trimer protein nanoparticles are superior immunogens compared to trimers alone for priming of broadly neutralizing antibody (bnAb) B cell lineages. The successful expression of complex multivalent nanoparticle immunogens with mRNAs has not been demonstrated. Here we show that mRNAs can encode antigenic Env trimers on ferritin nanoparticles that initiate bnAb precursor B cell expansion and induce serum autologous tier 2 neutralizing activity in bnAb precursor VH + VL knock-in mice. Next generation sequencing demonstrated acquisition of critical mutations, and monoclonal antibodies that neutralized heterologous HIV-1 isolates were isolated. Thus, mRNA-LNP can encode complex immunogens and are of use in design of germline-targeting and sequential boosting immunogens for HIV-1 vaccine development.


What is the case for care home medicine? The geriatrician's perspective.
This paper makes the case for a specific set of competencies required to adequately care for care home residents. It proposes that staff with these competencies need to work in an appropriate service context in order to achieve the best outcomes. These competencies and contextual factors are not currently recognised or championed by any single constituency within the existing healthcare infrastructure and there is a case that service delivery would be well served by recognition that 'care home medicine' is a distinct entity.


[Development and psychometric validation of a new screening questionnaire for erectile dysfunction (SQUED questionnaire)].
It is necessary to have simple tools to screen erectile dysfunction (ED) in an easy, reliable and valid manner. The objective of this study was to develop and validate a short diagnostic questionnaire for erectile dysfunction [SQUED), easy to use in the primary care setting. The development of SQUED included: concepts identification, item generation and evaluation of contents and face validity through interviews with subjects to assess comprehension and idiomatic adequacy. The psychometric validation was conducted in an epidemiologic, observational, comparative and multicenter study. Participants should complete the questionnaire in primary care setting and send to the specialist to confirm the diagnosis by an in depth interview and the application of IIEF questionnaire. Internal consistency, test-retest reliability and sensitivity and specificity of SQUED was evaluated. Out of 405 subjects enrolled, 316 (208 ED and 108 non-ED) were evaluable. The SQUED questionnaire showed a high internal consistency (Cronbach's alpha = 0.92) and a good test-retest reliability (Kappa index = 0.77). Furthermore, the questionnaire showed a good diagnostic capacity with high values of sensitivity and specificity, 0.87 and 0.78, respectively, in relation to the diagnosis made by urologists. The cutting point was established at a score of 12. The SQUED questionnaire is a simple, easy to use and reliable instrument. It can become a useful tool, and the shortest validated, for primary care physicians to easily screen for ED patients. Its simplicity should facilitate an easy cultural adaptation and validation into other languages.


Transforming Growth Factor-induced Protein Promotes NF-κB-mediated Angiogenesis during Postnatal Lung Development.
Pulmonary angiogenesis is a key driver of alveolarization. Our prior studies showed that NF-κB promotes pulmonary angiogenesis during early alveolarization. However, the mechanisms regulating temporal-specific NF-κB activation in the pulmonary vasculature are unknown. To identify mechanisms that activate proangiogenic NF-κB signaling in the developing pulmonary vasculature, proteomic analysis of the lung secretome was performed using two-dimensional difference gel electrophoresis. NF-κB activation and angiogenic function was assessed in primary pulmonary endothelial cells (PECs) and TGFBI (transforming growth factor-β-induced protein)-regulated genes identified using RNA sequencing. Alveolarization and pulmonary angiogenesis was assessed in wild-type and Tgfbi null mice exposed to normoxia or hyperoxia. Lung TGFBI expression was determined in premature lambs supported by invasive and noninvasive respiratory support. Secreted factors from the early alveolar, but not the late alveolar or adult lung, promoted proliferation and migration in quiescent, adult PECs. Proteomic analysis identified TGFBI as one protein highly expressed by the early alveolar lung that promoted PEC migration by activating NF-κB via αvβ3 integrins. RNA sequencing identified Csf3 as a TGFBI-regulated gene that enhances nitric oxide production in PECs. Loss of TGFBI in mice exaggerated the impaired pulmonary angiogenesis induced by chronic hyperoxia, and TGFBI expression was disrupted in premature lambs with impaired alveolarization. Our studies identify TGFBI as a developmentally regulated protein that promotes NF-κB-mediated angiogenesis during early alveolarization by enhancing nitric oxide production. We speculate that dysregulation of TGFBI expression may contribute to diseases marked by impaired alveolar and vascular growth.


Influence of environmental temperature on protein and amino acid needs of poultry.
Poultry production generally declines during adverse hot weather conditions. Dietary adjustments are often made to attempt to overcome the reduced performance, with an increase in dietary protein or amino acids being common. A review of the literature regarding the influence of environmental temperature on the amino acid needs of the chicken does not support an increase in the amino acid needs at temperature in excess of 32 C. Studies with broilers, growing pullets, and laying hens are in agreement that increased amino acid levels cannot effectively compensate for the reduced feed intake frequently observed at the higher envirnomental temperatures. Excessive dietary amino acid levels contribute to inhibition of feed intake and reduction in growth rate. Studies with practical-type diets have indicated that the performance of broiler chicks under conditions of high environmental temperatures may be improved through minimizing the excess levels of amino acids in the diet. This may be accomplished by means of blending protein sources combined with supplementation with commercially synthesized amino acids.


Complete pathological remission and tertiary lymphoid structures are associated with the efficacy of resectable NSCLC receiving neoadjuvant chemoimmunotherapy: A double-center retrospective study.
This study aimed to investigate the relationship between complete pathological remission (PCR), tertiary lymphoid structure (TLS) maturation and expression and clinical outcomes of patients with resectable non-small cell lung cancer (NSCLC) receiving neoadjuvant chemoimmunotherapy. Totally 80 patients with resectable NSCLC (stage IB-IIIB) receiving neoadjuvant chemoimmunotherapy were analyzed. We used the Kaplan-Meier method to plot survival curves and the log-rank test to compare differences. Among all patients included, 45 patients (56.25%) achieved major pathological response (MPR), including 30 patients (37.50%) with PCR. The proportion of patients diagnosed with stage IB, II, IIIA and IIIB was 1.25%, 10.00%, 52.50% and 36.25%, respectively. We divided patients into PCR group and non-PCR group respectively according to whether they achieved PCR. We found that patients achieving PCR had significantly improved disease-free survival (DFS) (mDFS: NR vs. 20.24 months, P = .020). TLS expression was low in 43 cases (53.75%) and high in 37 cases (46.25%). TLS maturation was low in 55 cases (68.75%) and high in 25 cases (31.25%). The DFS of patients with TLS high-maturation (34.07 vs. 22.30 months, P = .024) and TLS high-expression (34.07 vs. 22.30 months, P = .041) was significantly longer. In most subgroups, the PCR, TLS high-maturation and TLS high-expression group respectively achieved a better clinical outcome relative to the non-PCR, TLS low-maturation and TLS low-expression group. In patients with resectable NSCLC receiving neoadjuvant chemoimmunotherapy, the acquirement of PCR may predict better DFS. In addition, high expression and maturation of TLS may be prognostic factors.


The role of ethical banks in health care policy and financing in Spain.
Ethical, social, or civic banks, constitute a secondary source of financing, which is particularly relevant in Southern and Central Europe. However there is no information on the scientific literature on this source of health care financing. We review the characteristics of saving banks in Spain and illustrate the contribution of one institution "Obra Social Caixa Catalunya" (OS-CC) to the health care financing in Spain. Savings bank health care funding was equivalent to 3 percent of the public health expenditure for 2008. The programs developed by OS-CC illustrate the complex role of savings banks in health financing, provision, training, and policy, particularly in the fields of integrated care and innovation. Financing is a basic tool for health policy. However, the role of social banking in the development of integrated care networks has been largely disregarded, in spite of its significant contribution to complementary health and social care in Southern and Central Europe. Decision makers both at the public health agencies and at the social welfare departments of savings banks should become aware of the policy implications and impact of savings bank activities in the long-term care system.


Optimization of brain metabolism using metabolic-targeted therapeutic hypothermia can reduce mortality from traumatic brain injury.
Therapeutic hypothermia is widely used to treat traumatic brain injuries (TBIs). However, determining the best hypothermia therapy strategy remains a challenge. We hypothesized that reducing the metabolic rate, rather than reaching a fixed body temperature, would be an appropriate target because optimizing metabolic conditions especially the brain metabolic environment may enhance neurologic protection. A pilot single-blind randomized controlled trial was designed to test this hypothesis, and a nested metabolomics study was conducted to explore the mechanics thereof. Severe TBI patients (Glasgow Coma Scale score, 3-8) were randomly divided into the metabolic-targeted hypothermia treatment (MTHT) group, 50% to 60% rest metabolic ratio as the hypothermia therapy target, and the body temperature-targeted hypothermia treatment (BTHT) control group, hypothermia therapy target of 32°C to 35°C body temperature. Brain and circulatory metabolic pool blood samples were collected at baseline and on days 1, 3, and 7 during the hypothermia treatment, which were selected randomly from a subgroup of MTHT and BTHT groups. The primary outcome was mortality. Using H nuclear magnetic resonance technology, we tracked and located the disturbances of metabolic networks. Eighty-eight severe TBI patients were recruited and analyzed from December 2013 to December 2014, 44 each were assigned in the MTHT and BTHT groups (median age, 42 years; 69.32% men; mean Glasgow Coma Scale score, 6.17 ± 1.02). The mortality was significantly lower in the MTHT than the BTHT group (15.91% vs. 34.09%; p = 0.049). From these, eight cases of MTHT and six cases from BTHT group were enrolled for metabolomics analysis, which showed a significant difference between the brain and circulatory metabolic patterns in MTHT group on day 7 based on the model parameters and scores plots. Finally, metabolites representing potential neuroprotective monitoring parameters for hypothermia treatment were identified through H nuclear magnetic resonance metabolomics. MTHT can significantly reduce the mortality of severe TBI patients. Metabolomics research showed that this strategy could effectively improve brain metabolism, suggesting that reducing the metabolic rate to 50% to 60% should be set as the hypothermia therapy target. Therapeutic study, Level I.


Endogenous and diet-induced hypercholesterolemia in nonhuman primates: effects of age, adiposity, and diabetes on lipoprotein profiles.
Nonhuman primates (NHPs) share with humans many features of lipid metabolism and often develop all features of the metabolic syndrome, including hypertriglyceridemia and low high-density lipoprotein cholesterol, and have been used in many studies of potential therapeutics during the preclinical phase. Here we identify for the first time in middle-aged and older rhesus the natural occurrence of hypercholesterolemia, and this hypercholesterolemia develops despite maintenance on a low-cholesterol diet. The aims of this study were to (a) define normal and hypercholesterolemia in rhesus monkeys, (b) determine the factors associated with the development of hypercholesterolemia, (c) compare the lipoprotein profiles in adult rhesus monkeys fed a low-fat/low-cholesterol diet (LFLC) with the profiles of human subjects, and (d) determine the effect of a 16-week high-fat/high-cholesterol (HFHC) diet feeding on total cholesterol and lipoprotein profiles in middle-aged and older monkeys. In our colony, maintained on a constant diet with negligible cholesterol, the mean total cholesterol level in healthy nondiabetic monkeys was 3.7 ± 0.02 mmol/L, with hypercholesterolemia identified as the 95th percentile of the normal cholesterol distribution (≥5.2 mmol/L). Severe hypercholesterolemia developed in the HFHC-fed group; however, despite the high-fat diet composition, unexpectedly, no weight gain occurred in these NHPs. The diet-induced hypercholesterolemia differed significantly in lipoprotein pattern from that of the spontaneous hypercholesterolemia. In summary, despite ingesting only a LFLC, NHPs frequently develop hypercholesterolemia, reflecting lipoprotein patterns similar to human subjects; and this lipid profile of spontaneous hypercholesterolemia differs significantly from the hypercholesterolemia induced by an HFHC diet.


Determination of allergenicity to three cow's milk hydrolysates and an amino acid-derived formula in children with cow's milk allergy.
Products based on hydrolysed cow milk proteins or amino acid mixtures are recommended in children with cow's milk hypersensitivity. However, some children who are allergic to cow's milk and who clinically react to substitute milk formulas have been observed. To determine the tolerance and allergenicity of protein hydrolysate or amino acid-derived formulas in children with IgE-mediated cow's milk allergy. Twenty children with positive cow's milk challenges, positive skin prick tests and/or serum-specific IgE antibodies to cow's milk were selected. Oral challenges, skin prick tests and serum-specific IgE antibodies to extensively hydrolysed whey formula, partially hydrolysed whey formula, extensively hydrolysed casein formula and amino acid-derived formula were performed. Five out of 17 (5/17) children reacted to partially hydrolysed whey formula, (3/16) to extensively hydrolysed whey formula, (2/10) to amino acid-derived formula, (1/16) to extensively hydrolysed casein formula. Only extensively hydrolysed casein formula was tolerated by at least 90% (with 95% confidence intervals) of children. Hydrolysates provoked early and delayed clinical reactions, amino acid mixtures only delayed reactions. Partially hydrolysed whey formula elicited a significantly higher number of positive skin prick test reactions than other formulas. Two children had specific IgE antibodies to extensively hydrolysed whey formula, one to partially hydrolysed whey formula, one to extensively hydrolysed casein formula and none to amino acid-derived formula. In this study, none of the cow's milk substitutes has been found to be non-allergenic. Our results suggest that in children with IgE-mediated cow's milk allergy, the first ingestions of extensively hydrolysed cow's milk protein formulas require strict medical supervision because of immediate reactions. This is not the case for amino acid-derived formula. Moreover, our data suggest that treatment of children allergic to cow's milk with cow's milk substitutes should be monitored for several days to document tolerance.


Evaluation of silver nanoparticles for the prevention of SARS-CoV-2 infection in health workers: In vitro and in vivo.
SARS-CoV-2 infection in hospital areas is of a particular concern, since the close interaction between health care personnel and patients diagnosed with COVID-19, which allows virus to be easily spread between them and subsequently to their families and communities. Preventing SARS-CoV-2 infection among healthcare personnel is essential to reduce the frequency of infections and outbreaks during the pandemic considering that they work in high-risk areas. In this research, silver nanoparticles (AgNPs) were tested in vitro and shown to have an inhibitory effect on SARS-CoV-2 infection in cultured cells. Subsequently, we assess the effects of mouthwash and nose rinse with ARGOVIT® silver nanoparticles (AgNPs), in the prevention of SARS-CoV-2 contagion in health workers consider as high-risk group of acquiring the infection in the General Tijuana Hospital, Mexico, a hospital for the exclusive recruitment of patients diagnosed with COVID-19. We present a prospective randomized study of 231 participants that was carried out for 9 weeks (during the declaration of a pandemic). The "experimental" group was instructed to do mouthwash and nose rinse with the AgNPs solution; the "control" group was instructed to do mouthwashes and nose rinse in a conventional way. The incidence of SARS-CoV-2 infection was significantly lower in the "experimental" group (two participants of 114, 1.8%) compared to the "control" group (thirty-three participants of 117, 28.2%), with an 84.8% efficiency. We conclude that the mouth and nasal rinse with AgNPs helps in the prevention of SARS-CoV-2 infection in health personnel who are exposed to patients diagnosed with COVID-19.


Amelogenesis imperfecta: functional and esthetic restoration of a severely compromised dentition.
The treatment of patients with severe forms of amelogenesis imperfecta presents an interesting challenge to the dental team. A 16-year-old girl presented with a severely compromised occlusion and poor esthetics that had lowered her self-esteem. Preoperative investigations included the determination of completion of growth, an accurate diagnostic waxup, and an evaluation of clinical crown lengths. Periodontal full-flap surgery was planned to lengthen clinical crowns and create gingival harmony. This was to be followed by placement of 28 complete crowns to restore occlusal function and gain a pleasing esthetic result. All-porcelain crowns were placed from premolar to premolar in each arch (20 crowns), and porcelain-fused-to-metal crowns were placed on each molar tooth in each arch (8 crowns). The final treatment result provided this patient with a mutually protective occlusion and with esthetics that greatly enhanced her self-image.


Protonated dipeptide losses from b(5) and b(4) ions of side chain hydroxyl group containing pentapeptides.
In this study, C-terminal protonated dipeptide eliminations were reported for both b5 and b4 ions of side chain hydroxyl group (-OH) containing pentapeptides. The study utilized the model C-terminal amidated pentapeptides having sequences of XGGFL and AXVYI, where X represents serine (S), threonine (T), glutamic acid (E), aspartic acid (D), or tyrosine (Y) residue. Upon low-energy collision-induced dissociation (CID) of XGGFL (where X = S, T, E, D, and Y) model peptide series, the ions at m/z 279 and 223 were observed as common fragments in all b5 and b4 ion (except b4 ion of YGGFL) mass spectra, respectively. By contrast, peptides, namely SMeGGFL-NH2 and EOMeGGFL-NH2, did not show either the ion at m/z 279 or the ion at m/z 223. It is shown that the side chain hydroxyl group is required for the possible mechanism to take place that furnishes the protonated dipeptide loss from b5 and b4 ions. In addition, the ions at m/z 295 and 281 were detected as common fragments in all b5 and b4 ion (except b4 ion of AYVYI) mass spectra, respectively, for AXVYI model peptide series. The MS(4) experiments exhibited that the fragment ions at m/z 279, 223, 295, and 281 entirely reflect the same fragmentation behavior of [M + H](+) ion generated from commercial dipeptides FL-OH, GF-OH, YI-OH, and VY-OH. These novel eliminations reported here for b5 and b4 ions can be useful in assigning the correct and reliable peptide sequences for high-throughput proteomic studies.


3D-printed bioactive and biodegradable hydrogel scaffolds of alginate/gelatin/cellulose nanocrystals for tissue engineering.
The 3D-printed hybrid biodegradable hydrogels composed of alginate, gelatin, and cellulose nanocrystals (CNCs) were prepared to provide a favorable environment for cell proliferation, adhesion, nutrients exchange, and matrix mineralization for bone tissue engineering (BTE) applications. The hybrid scaffolds exhibited enhanced mechanical strength compared to the pure polymer scaffolds. The biocompatibility, differentiation potential, and bone regeneration potential of the printed scaffolds were evaluated by DAPI staining, live-dead assay, alizarin Red-S (ARS) staining, real-time PCR (qRT-PCR), and μCT analysis, respectively. Enhanced cell proliferation has occurred 1% CNC/Alg/Gel scaffolds compared to the control. The cells were adequately adhered to the scaffold and exhibited the flattened structure. Improved mineralization was observed in the 1% CNC/Alg/Gel scaffolds' presence than the control, showing their mineralization efficiency. A significant enhancement in the expression of osteogenic-specific gene markers (Runx2, ALP, BMP-2, OCN, OPN, BSP, and COL1) has occurred with 1% CNC/Alg/Gel than the control, indicating their osteogenic potential. Furthermore, enhanced bone formation was observed in the scaffolds treated groups than the control in the calvaria critical-sized defects (CCD-1) model, suggesting their improved bone regeneration potential. Therefore, the fabricated scaffolds have the potential to explore as a biomaterial for tissue engineering.


Genome-wide DNA polymorphisms in four Actinidia arguta genotypes based on whole-genome re-sequencing.
Among the genus Actinidia, Actinidia arguta possesses the strongest cold resistance and produces fresh fruit with an intense flavor. To investigate genomic variation that may contribute to variation in phenotypic traits, we performed whole-genome re-sequencing of four A. arguta genotypes originating from different regions in China and identified the polymorphisms using InDel markers. In total, 4,710,650, 4,787,750, 4,646,026, and 4,590,616 SNPs and 1,481,002, 1,534,198, 1,471,304, and 1,425,393 InDels were detected in the 'Ruby-3', 'Yongfeng male', 'Kuilv male', and 'Hongbei male' genomes, respectively, compared with the reference genome sequence of cv 'Hongyang'. A subset of 120 InDels were selected for re-sequencing validation. Additionally, genes related to non-synonymous SNPs and InDels in coding domain sequences were screened for functional analysis. The analysis of GO and KEGG showed that genes involved in cellular responses to water deprivation, sucrose transport, decreased oxygen levels and plant hormone signal transduction were significantly enriched in A. arguta. The results of this study provide insight into the genomic variation of kiwifruit and can inform future research on molecular breeding to improve cold resistance in kiwifruit.


Profound alterations of intestinal microbiota following a single dose of clindamycin results in sustained susceptibility to Clostridium difficile-induced colitis.
Antibiotic-induced changes in the intestinal microbiota predispose mammalian hosts to infection with antibiotic-resistant pathogens. Clostridium difficile is a Gram-positive intestinal pathogen that causes colitis and diarrhea in patients following antibiotic treatment. Clindamycin predisposes patients to C. difficile colitis. Here, we have used Roche-454 16S rRNA gene pyrosequencing to longitudinally characterize the intestinal microbiota of mice following clindamycin treatment in the presence or absence of C. difficile infection. We show that a single dose of clindamycin markedly reduces the diversity of the intestinal microbiota for at least 28 days, with an enduring loss of ca. 90% of normal microbial taxa from the cecum. Loss of microbial complexity results in dramatic sequential expansion and contraction of a subset of bacterial taxa that are minor contributors to the microbial consortium prior to antibiotic treatment. Inoculation of clindamycin-treated mice with C. difficile (VPI 10463) spores results in rapid development of diarrhea and colitis, with a 4- to 5-day period of profound weight loss and an associated 40 to 50% mortality rate. Recovering mice resolve diarrhea and regain weight but remain highly infected with toxin-producing vegetative C. difficile bacteria and, in comparison to the acute stage of infection, have persistent, albeit ameliorated cecal and colonic inflammation. The microbiota of "recovered" mice remains highly restricted, and mice remain susceptible to C. difficile infection at least 10 days following clindamycin, suggesting that resolution of diarrhea and weight gain may result from the activation of mucosal immune defenses.


Paradoxical cerebral arterial embolization of cement during intraoperative vertebroplasty: case report.
Paradoxical cerebral embolism of cement occurred in a 78-year-old woman after cement-assisted transpedicular spinal fixation surgery. Multiple pulmonary emboli of polymethylmethacrylate precipitated pulmonary hypertension and right-to-left shunting into the systemic circulation through a patent foramen ovale. This rare complication occurred because of failure to recognize venous migration of cement during the procedure and the injection of multiple levels in one setting. Although this was an open procedure, the technical aspects were the same as for vertebroplasty and the precautions should be applied to percutaneous vertebroplasty.


The 'Italian Experience' reconsidered.
Our analysis of the problems of the Italian psychiatric services has drawn a lively correspondence challenging our findings. In September 1985, we undertook a second study tour in Italy. This is now reported, with notes on specific criticisms of the earlier paper.


Processing of double-R-loops in (CAG)·(CTG) and C9orf72 (GGGGCC)·(GGCCCC) repeats causes instability.
R-loops, transcriptionally-induced RNA:DNA hybrids, occurring at repeat tracts (CTG)n, (CAG)n, (CGG)n, (CCG)n and (GAA)n, are associated with diseases including myotonic dystrophy, Huntington's disease, fragile X and Friedreich's ataxia. Many of these repeats are bidirectionally transcribed, allowing for single- and double-R-loop configurations, where either or both DNA strands may be RNA-bound. R-loops can trigger repeat instability at (CTG)·(CAG) repeats, but the mechanism of this is unclear. We demonstrate R-loop-mediated instability through processing of R-loops by HeLa and human neuron-like cell extracts. Double-R-loops induced greater instability than single-R-loops. Pre-treatment with RNase H only partially suppressed instability, supporting a model in which R-loops directly generate instability by aberrant processing, or via slipped-DNA formation upon RNA removal and its subsequent aberrant processing. Slipped-DNAs were observed to form following removal of the RNA from R-loops. Since transcriptionally-induced R-loops can occur in the absence of DNA replication, R-loop processing may be a source of repeat instability in the brain. Double-R-loop formation and processing to instability was extended to the expanded C9orf72 (GGGGCC)·(GGCCCC) repeats, known to cause amyotrophic lateral sclerosis and frontotemporal dementia, providing the first suggestion through which these repeats may become unstable. These findings provide a mechanistic basis for R-loop-mediated instability at disease-associated repeats.


Extraction efficiency of an extraction buffer used to quantify lysozyme deposition on conventional and silicone hydrogel contact lens materials.
Extracting lysozyme from Food and Drug Administration group IV etafilcon lenses by using 0.2% trifluoroacetic acid and acetonitrile (TFA/ACN) is a well-established procedure. TFA/ACN has been the extraction buffer of choice for extracting proteins from silicone hydrogel contact lenses. The purpose of this study was to determine the efficiency of TFA/ACN in extracting lysozyme from silicone hydrogel and etafilcon lenses by using an in vitro model. ACUVUE 2, Focus NIGHT & DAY, O2 Optix, PureVision, and ACUVUE Advance lenses were incubated in simple lysozyme solution and a complex artificial tear solution consisting of multiple tear components containing lysozyme labeled with iodine 125. All the silicone hydrogel lenses were incubated for 28 days, whereas the ACUVUE 2 lenses were incubated for 7 days at 37 degrees C with constant rotation. After the incubation period, radioactive counts were determined, and the lenses were placed in an appropriate volume of the buffer for 24 hours in darkness. The lenses were removed from the buffer, and radioactive counts were determined again. Extraction efficiencies for lysozyme from the artificial tear solution were 97.2% +/- 1.2% for ACUVUE 2, 64.3% +/- 6.2% for Focus NIGHT & DAY, 62.5% +/- 5.6% for O2 Optix, 53.5% +/- 5.8% for PureVision, and 89.2% +/- 3.4% for ACUVUE Advance. Results were similar for the lysozyme extracted after incubating in the simple lysozyme solution. TFA/ACN is extremely efficient at extracting lysozyme deposited on etafilcon lenses. However, it does not extract all the lysozyme deposited on silicone hydrogel lenses, and alternative extraction procedures should be sought.


Marginal Bone Loss Around Early-Loaded SLA and SLActive Implants: Radiological Follow-Up Evaluation Up to 6.5 Years.
The aim of this study was to compare marginal bone loss around early-loaded SLA and SLActive tissue-level implants (Straumann Dental Implants; Institut Straumann AG, Basel, Switzerland) after a mean of 81-month follow-up period. One hundred seven SLA and 68 SLActive implants were placed in 55 patients and loaded with final restoration after 8 and 3 weeks of healing time, respectively. Marginal bone loss around implants was determined radiographically at initial and after a mean observation time ranging between 20 and 81 months. The effect of location (mandible vs maxilla), smoking habit, sex, implant length and diameter, and the type of prosthesis on the marginal bone loss was evaluated. The overall cumulative survival rate was 98.2% being 99% for SLA implants and 97% for SLActive implants. After 20-month follow-up period, mean marginal bone loss values for the SLA and SLActive implants were 0.24 and 0.17 mm, respectively. After 81 months, mean marginal bone loss for the SLA and SLActive implants reached 0.71 and 0.53 mm, respectively. Marginal bone loss was affected by the length and type of implant and patients' smoking habit after a mean observation time of 20 months. However, none of the parameters had any significant effect on the marginal bone loss after a follow-up period of 81 months. With both SLA and SLActive implants, successful clinical results could be achieved up to 6.5 years of follow-up period.


Disparities in health care determine prognosis in newly diagnosed glioblastoma.
OBJECTIVE Glioblastoma (GBM) is an aggressive brain malignancy with a short overall patient survival, yet there remains significant heterogeneity in outcomes. Although access to health care has previously been linked to impact on prognosis in several malignancies, this question remains incompletely answered in GBM. METHODS This study was a retrospective analysis of 354 newly diagnosed patients with GBM who underwent first resection at the authors' institution (2007-2015). RESULTS Of the 354 patients (median age 61 years, and 37.6% were females), 32 (9.0%) had no insurance, whereas 322 (91.0%) had insurance, of whom 131 (40.7%) had Medicare, 45 (14%) had Medicaid, and 146 (45.3%) had private insurance. On average, insured patients survived almost 2-fold longer (p < 0.0001) than those who were uninsured, whereas differences between specific insurance types did not influence survival. The adjusted hazard ratio (HR) for death was higher in uninsured patients (HR 2.27 [95% CI 1.49-3.33], p = 0.0003). Age, mean household income, tumor size at diagnosis, and extent of resection did not differ between insured and uninsured patients, but there was a disparity in primary care physician (PCP) status-none of the uninsured patients had PCPs, whereas 72% of insured patients had PCPs. Postoperative adjuvant treatment rates with temozolomide (TMZ) and radiation therapy (XRT) were significantly less in uninsured (TMZ in 56.3%, XRT in 56.3%) than in insured (TMZ in 75.2%, XRT in 79.2%; p = 0.02 and p = 0.003) patients. Insured patients receiving both agents had better prognosis than uninsured patients receiving the same treatment (9.1 vs 16.34 months; p = 0.025), suggesting that the survival effect in insured patients could only partly be explained by higher treatment rates. Moreover, having a PCP increased survival among the insured cohort (10.7 vs 16.1 months, HR 1.65 [95% CI 1.27-2.15]; p = 0.0001), which could be explained by significant differences in tumor diameter at initial diagnosis between patients with and without PCPs (4.3 vs 4.8 cm, p = 0.003), and a higher rate of clinical trial enrollment, suggesting a critical role of PCPs for a timelier diagnosis of GBM and proactive cancer care management. CONCLUSIONS Access to health care is a strong determinant of prognosis in newly diagnosed patients with GBM. Any type of insurance coverage and having a PCP improved prognosis in this patient cohort. Higher rates of treatment with TMZ plus XRT, clinical trial enrollment, fewer comorbidities, and early diagnosis may explain survival disparities. Lack of health insurance or a PCP are major challenges within the health care system, which, if improved upon, could favorably impact the prognosis of patients with GBM.


Obstructive Jaundice as a Complication of Macrocystic Serous Cystadenoma of the Pancreas.
Macrocystic serous cystadenoma is an unusual and essentially benign pancreatic tumor. Herein, we report on a 40-year-old woman diagnosed with macrocystic serous cystadenoma who presented with obstructive jaundice. A cystic lesion in the head and body of the pancreas was revealed by abdominal computed tomography. Intraoperative pancreatic cyst aspiration ruled out mucinous cystic neoplasm which has a malignant potential. The pancreatic cyst fluid cytology was basophilic amorph materials concluded as benign cystic lesion. Internal drainage was performed instead of pancreatic resection which showed good outcome. Biliary obstruction is a rare complication of serous cystadenoma. This case describes an unusual clinical presentation of macrocystic serous cystadenoma.


Endoscopic removal of an incidentally discovered intrauterine contraceptive device eroding into the rectum.
Intrauterine contraceptive device (IUCD) is a common birth control method. It is safe but can be associated with serious complications including migration into the peritoneal cavity and penetration into other intra-abdominal and pelvic viscera; most commonly the rectosigmoid colon. Different retrieval methods including endoscopy, laparoscopy or open abdominal surgery have been described. We report the case of 38-year-old woman who became pregnant shortly after insertion of the IUCD 6 years prior to presentation. She delivered vaginally and 'expulsion' of the device was assumed. Some 4 years later, she had another IUCD inserted and remained asymptomatic till she recently presented with iron-deficiency anaemia. As part of the investigation, diagnostic colonoscopy was performed. Surprisingly, the old IUCD was found penetrating into the midrectum. Uneventful endoscopic removal was performed and she remained well at 3-month follow-up. Migrating IUCD remains asymptomatic and may be discovered accidentally during routine investigation for some other symptoms.


Low-drift-rate external cavity diode laser.
We present the design, construction, and simulation of a simple, low-cost external cavity diode laser with a measured free-running frequency drift rate of 1.4(1) MHz/h at 852 nm. This performance is achieved in a compact aluminum structure held inside an airtight, temperature-controlled enclosure. The high thermal conductivity of the laser cavity and the stable temperature environment inside the enclosure minimize the time-varying, spatial temperature gradients across the laser cavity. We present thermal finite element method simulations, which quantify the effects of temperature gradients, and suggest that the drift rate is likely limited by the laser-diode and piezo-aging.


Nanodiamond-PMO for two-photon PDT and drug delivery.
In this article, we highlight the properties of nanodiamonds (ND), which were encapsulated in periodic mesoporous organosilica nanoparticles (PMO) and were able to generate reactive oxygen species for photodynamic applications upon two-photon excitation (TPE). The ND@PMO nanoparticles were characterized by various techniques and were then loaded with the anti-cancer drug doxorubicin. The release of the drug was pH sensitive and a synergistic cancer cell killing effect was observed when cancer cells were incubated with doxorubicin-loaded ND@PMO and irradiated with two-photon excitation at 800 nm.


Multilevel confirmatory factor analysis of the Effective Behavior Support Self-Assessment Survey.
One evaluation measure available through the Positive Behavioral Interventions and Supports framework is the Effective Behavior Support Self-Assessment Survey (SAS). Evaluations of the SAS have supported its factor structure. However, the SAS is designed to be completed by school personnel who are nested within other levels of analysis (e.g., schools, grade level, district). There have been no studies examining the SAS from a multilevel perspective. The current study addressed this gap by evaluating the SAS using data from 1,726 respondents across 36 public schools in 3 school districts. Another purpose of this study was to evaluate whether the SAS can be shortened yet remain psychometrically sound to encourage completion of the survey by busy school staff members. Intraclass correlation coefficients indicated substantial dependence in item responses. Factor correlations were substantial across all models tested. Compared with a 2nd-order, hierarchical model and the basic 4-factor model based on the original hypothesized structure of the SAS, the best combination of fit and parsimony was obtained from a reduced set of 24 of the original 46 items organized within a multilevel bifactor structure (e.g., comparative fit index = .979, root-mean-square error of approximation = .019), suggesting that the SAS may be shortened. The bifactor model partitioned item variances to a general implementation fidelity factor and to 1 of the 4 hypothesized factors. These results suggest that the SAS items aim to describe dimensions for measuring implementation status that may be as relevant at the school level as they are at the individual level. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Classification analysis of the transcriptosome of nonlesional cultured dermal fibroblasts from systemic sclerosis patients with early disease.
To compare the transcriptosome of early-passage nonlesional dermal fibroblasts from systemic sclerosis (SSc) patients with diffuse disease and matched normal controls in order to gain further understanding of the gene activation patterns that occur in early disease. Total RNA was isolated from early-passage fibroblasts obtained from nonlesional skin biopsy specimens from 21 patients with diffuse SSc (disease duration <5 years in all but 1) and 18 healthy controls who were matched to the cases by age (+/-5 years), sex, and race. Array experiments were performed on a 16,659-oligonucleotide microarray utilizing a reference experimental design. Supervised methods were used to select differentially expressed genes. Quantitative polymerase chain reaction (PCR) was used to independently validate the array results. Of the 8,324 genes that passed filtering criteria, classification analysis revealed that <5% were differentially expressed between SSc and normal fibroblasts. Individually, differentially expressed genes included COL7A1, COL18A1 (endostatin), DAF, COMP, and VEGFB. Using the panel of genes discovered through classification analysis, a set of model predictors that achieved reasonably high predictive accuracy was developed. Analysis of 1,297 gene ontology (GO) classes revealed 35 classes that were significantly dysregulated in SSc fibroblasts. These GO classes included anchoring collagen (30934), extracellular matrix structural constituent (5201), and complement activation (6958, 6956). Validation by quantitative PCR demonstrated that 7 of 7 genes selected were concordant with the array results. Fibroblasts cultured from nonlesional skin of patients with SSc already have detectable abnormalities in a variety of genes and cellular processes, including those involved in extracellular matrix formation, fibrillogenesis, complement activation, and angiogenesis.


Lower level of serum potassium and higher level of C-reactive protein as an independent risk factor for giant aneurysms in Kawasaki disease.
Giant aneurysms are the most serious issue of patients with Kawasaki disease (KD). To clarify risk factors for these giant aneurysms, we conducted a matched case-control study. Among the patients reported in nationwide surveys, 117 patients with giant aneurysms had an unequivocal new diagnosis and presented at the treatment center within 9 d of illness. We obtained clinical information on admission of about 69 patients (case) from the treatment centers. One control was selected for each case, an age- and sex-matched patient without coronary involvement, reported from the same treatment center at about the same time as the case, and we obtained the same clinical information about controls. Fourteen variables were analysed with a conditional logistic regression model: body temperature, hematocrit, hemoglobin, numbers of leukocyte and platelets, concentrations of serum albumin, globulin, total cholesterol, sodium, potassium and chloride, erythrocyte sedimentation rate, C-reactive protein and alanine aminotransferase activity. After adjustment for age, duration of illness before admission and use of intravenous gamma globulin therapy, C-reactive protein [odds ratio (OR)=1.142, 95% confidence interval (CI) 1.054-1.237], alanine aminotransferase activity (OR=1.008, 95% CI 1.002-1.014), serum sodium concentration (OR=0.877, 95% CI 0.770-0.999) and serum potassium concentration (OR=0.319, 95% CI 0.124-0.822) were significantly related to the risk for giant aneurysms. Further analyses with these four explanatory variables revealed that C-reactive protein (OR=1.159, 95% CI 1.022-1.315) and serum potassium concentration (OR=0.222, 95% CI 0.052-0.948) met the significant level. Thus, the values for serum C-reactive protein and potassium are independent risk factors for the development of the giant aneurysms of Kawasaki disease.


[Directions of improvement of psycho-physiological supply of professional activity of specialists of manipulation of spacecrafts].
Were determined components of systems of guarantee of security of activity of specialists of manipulation of spacecrafts (SC): abidance by general organizational-methodological rules and principles of work, excluding violations in organization of duty, preventing of nonfulfillment and failure of sessions of control by SC, determination and deleting negative factors of operative activity, guarantee of safety of system "man-mechanism-environment." Were determined peculiarities of activity of specialists of manipulation of spacecrafts, their professionally important qualities, which had to be considered in effectuating of professional psycho-selection. Were shown regularities of changes in functional state and professional working capacity of operators of command-measured complexes during daily duty and during cycle of a year. Was made a conclusion about necessity of improvement of system of psychophysiological accompaniment of activity of specialists of manipulation of SC.


Acute Kidney Injury in Pediatric Patients: Diagnosis and Management in the Emergency Department.
Pediatric acute kidney injury is a condition that is underdiagnosed among children seen in the emergency department, and it has been associated with significant morbidity and mortality, including increased risk for chronic kidney disease. The most common etiologies in pediatric patients are now known to be due to hypovolemia, sepsis, shock, and cardiac dysfunction. This issue compares 3 classification systems for the diagnosis and staging of acute kidney injury and reviews the etiologies that lead to kidney injury in children. The management of pediatric acute kidney injury focuses on identifying patients at high risk, monitoring intravascular volume status, avoiding nephrotoxic medication exposure, and involving a pediatric nephrologist once acute kidney injury is diagnosed.


Association of serum IgG N-glycome and transforming growth factor-β1 with hepatitis B virus e antigen seroconversion during entecavir therapy.
Aberrant serum IgG N-glycome has been demonstrated in various autoimmune diseases and viral infections. However, the correlation between serum IgG N-glycome and cytokine is unclear. In addition, the clinical relevance of IgG glycosylation and cytokine changes in the treatment outcome of chronic hepatitis B (CHB) has never been assessed. One hundred and three treatment-naive patients with CHB and 101 healthy controls were enrolled in this retrospective cohort study. Serum samples in patients before and after 48weeks of entecavir treatment were collected. In-gel trypsinized serum IgG heavy chain was analyzed using liquid chromatography-tandem mass spectrometry. Selected ion chromatograms corresponding to 10 N-glycoforms on asparagine 297 were individually extracted to calculate the percentage of each glycoforms. Serum cytokine profiles were examined using enzyme-linked immunosorbent assay. Forty-eight weeks of entecavir treatment resulted in decreases in galactose-deficient (total G0) IgG and transforming growth factor (TGF)-β1 levels (both P<0.001) in patients with CHB. The changes in TGF-β1 (ΔTGF-β1) and IgG total G0 (Δtotal G0) levels during treatment were significantly correlated (r=0.403, P<0.001). Furthermore, higher levels of Δtotal G0 (P<0.01) and ΔTGF-β1 (P<0.001) were found in hepatitis B virus e antigen (HBeAg)-positive patients than in HBeAg-negative patients and were also found in patients with HBeAg seroconversion at week 48. The area under the receiver operating characteristic (ROC) curves for Δtotal G0 and ΔTGF-β1 to discriminate a week-48 HBeAg seroconversion were 0.835 and 0.830, respectively. These results suggested a correlation between serum cytokine and IgG N-glycome and its effect on the outcome of entecavir treatment in patients with CHB.


Biotin status of epileptics.
Microbiologically determined plasma biotin levels in 404 epileptics under long-term treatment with anticonvulsants were markedly lower than in 112 controls (p less than 0.0005). Patients with partial epilepsy had lower biotin levels and higher average daily intake of AC than those with generalized epilepsy. Epileptics treated with valproate sodium in monotherapy showed considerably higher biotin levels than epileptics with monotherapy of primidone (PRM), carbamazepine (CBZ), phenytoin (PHT) or phenobarbital (PB). The group of epileptics with high average daily dose of anticonvulsants had lower biotin levels than the group with low dose. In three patients with newly recognized epilepsy biotin levels were normal before starting anticonvulsant medication, increased during the first week and fell under the starting level in the following weeks. Four epileptics treated with PHT, PB, PRM or CBZ had an increased urinary excretion of organic acids, as found in patients with a deficiency of biotin-dependent carboxylases. In 37 epileptics undergoing long-term treatment plasma lactate concentrations were determined; they had a higher mean concentration than that found in controls. Our results suggest, that the lowering of biotin in epileptics is caused by intake of anticonvulsants and has a biochemical effect in these patients. It is discussed, whether this could be a factor in the mode of action of anticonvulsants.


Interactions of Neutrophils with the Polymeric Molecular Components of the Biofilm Matrix in the Context of Implant-Associated Bone and Joint Infections.
In the presence of orthopedic implants, opportunistic pathogens can easily colonize the biomaterial surfaces, forming protective biofilms. Life in biofilm is a central pathogenetic mechanism enabling bacteria to elude the host immune response and survive conventional medical treatments. The formation of mature biofilms is universally recognized as the main cause of septic prosthetic failures. Neutrophils are the first leukocytes to be recruited at the site of infection. They are highly efficient in detecting and killing planktonic bacteria. However, the interactions of these fundamental effector cells of the immune system with the biofilm matrix, which is the true interface of a biofilm with the host cells, have only recently started to be unveiled and are still to be fully understood. Biofilm matrix macromolecules consist of exopolysaccharides, proteins, lipids, teichoic acids, and the most recently described extracellular DNA. The latter can also be stolen from neutrophil extracellular traps (NETs) by bacteria, who use it to strengthen their biofilms. This paper aims to review the specific interactions that neutrophils develop when they physically encounter the matrix of a biofilm and come to interact with its polymeric molecular components.


Reduced renal blood flow in early cisplatin-induced acute renal failure in the rat.
Studies were designed to determine the cause of the reduced glomerular filtration rate (GFR) in early cisplatin-induced acute renal failure. Rats were studied 72 h following a single intraperitoneal injection of cisplatin (5 mg/kg) or vehicle (0.9% NaCl). Whole kidney GFR and blood flow were lower in cisplatin-treated animals than in controls (0.30 +/- 0.06 vs. 1.17 +/- 0.06 ml X min-1 X g kidney wt-1 and 5.30 +/- 0.62 vs. 8.25 +/- 0.43 ml X min-1 X g kidney wt-1, respectively; P less than 0.001), as were superficial nephron GFR and stop-flow pressure (20.2 +/- 2.1 vs. 34.5 +/- 2.0 nl X min-1 X g kidney wt-1 and 29.0 +/- 1.9 vs. 39.8 +/- 1.3 mmHg, respectively; P less than 0.001). After volume expansion, renal plasma flow increased in control rats, whereas whole kidney and single nephron GFR did not change. In experimental animals, whole kidney filtration rate rose to 0.58 +/- 0.07 ml X min-1 X g kidney wt-1, single nephron filtration rate increased to 29.9 +/- 3.5 nl X min-1 X g kidney wt-1 (P less than 0.005), and renal plasma flow increased to 5.62 +/- 0.60 ml X min-1 X g kidney wt-1 (P less than 0.05). Intratubular hydrostatic pressure was not different in the two groups before or after volume expansion. The results of these studies show that the reduced GFR in early cisplatin-induced renal failure is due, in part, to reversible changes in renal blood flow and renal vascular resistance.


Evaluation of plant seed DNA and botanical evidence for potential forensic applications.
Seeds, the reproductive organs of plants, are common as trace evidence from crime scenes. Seed evidence could be grouped into several categories based on the types of crimes they are associated with, including child abuse, homicides and drugs. Most commonly, seeds are examined microscopically and identified to the plant species level to show a linkage between persons and places. More recently, forensic researchers have evaluated the potential for extracting and typing DNA from seeds to further individualize the samples. As a model system, tomato seeds were examined microscopically after different cooking treatments and assessed for the potential to DNA type seeds for variety identification. A sufficient quantity and quality of DNA were recovered from uncooked, digested and undigested tomato seeds for amplified fragment length polymorphism (AFLP) analysis; however, any form of cooking destroyed the seed DNA. A simple microscopic analysis was able to distinguish between a cooked tomato seed versus an uncooked seed. This article is intended to provide an overview of case examples and current techniques for the forensic examination of seeds as plant-derived evidence.


Assessment of detailed conformations suggests strategies for improving cryoEM models: Helix at lower resolution, ensembles, pre-refinement fixups, and validation at multi-residue length scale.
We find that the overall quite good methods used in the CryoEM Model Challenge could still benefit greatly from several strategies for improving local conformations. Our assessments primarily use validation criteria from the MolProbity web service. Those criteria include MolProbity's all-atom contact analysis, updated versions of standard conformational validations for protein and RNA, plus two recent additions: first, flags for cis-nonPro and twisted peptides, and second, the CaBLAM system for diagnosing secondary structure, validating Cα backbone, and validating adjacent peptide CO orientations in the context of the Cα trace. In general, automated ab initio building of starting models is quite good at backbone connectivity but often fails at local conformation or sequence register, especially at poorer than 3.5 Å resolution. However, we show that even if criteria (such as Ramachandran or rotamer) are explicitly restrained to improve refinement behavior and overall validation scores, automated optimization of a deposited structure seldom corrects specific misfittings that start in the wrong local minimum, but just hides them. Therefore, local problems should be identified, and as many as possible corrected, before starting refinement. Secondary structures are confusing at 3-4 Å but can be better recognized at 6-8 Å. In future model challenges, specific steps being tested (such as segmentation) and the required documentation (such as PDB code of starting model) should each be explicitly defined, so competing methods on a given task can be meaningfully compared. Individual local examples are presented here, to understand what local mistakes and corrections look like in 3D, how they probably arise, and what possible improvements to methodology might help avoid them. At these resolutions, both structural biologists and end-users need meaningful estimates of local uncertainty, perhaps through explicit ensembles. Fitting problems can best be diagnosed by validation that spans multiple residues; CaBLAM is such a multi-residue tool, and its effectiveness is demonstrated.


The HOPE (Heart Outcomes Prevention Evaluation) Study: the design of a large, simple randomized trial of an angiotensin-converting enzyme inhibitor (ramipril) and vitamin E in patients at high risk of cardiovascular events. The HOPE study investigators.
To describe the design of the HOPE (Heart Outcomes Prevention Evaluation) study. Description of the key design features of HOPE, a large, simple randomized trial of two widely applicable treatments--ramipril, an angiotensin-converting enzyme inhibitor; and vitamin E, a naturally occurring antioxidant vitamin--in the prevention of myocardial infarction, stroke or cardiovascular death. Two-hundred and sixty-seven hospitals, physician offices and clinics in Canada, the United States, Mexico, Europe and South America. Over 9000 women and men aged 55 years and above at high risk for cardiovascular events such as myocardial infarction and stroke were recruited over 18 months. A 2X2 factorial design with ramipril and vitamin E with follow-up for up to four years. HOPE will be one of the largest trials of two new interventions to prevent myocardial infarction, stroke or cardiovascular death in high risk patients. The results of HOPE will have direct public health impact and are likely to be readily incorporated into clinical practice. Key design features of HOPE are inclusion of individuals at high risk of cardiovascular disease, inclusion of a substantial proportion of patients with diabetes (36%) and women (27%), and detailed substudies to provide data on mechanisms of benefit.


Effects of Repeated, Long-Duration Hyperoxic Water Immersions on Neuromuscular Endurance in Well-Trained Males.
This study examined the effects of repeated long-duration hyperoxic water immersions (WIs) at 1.35 atmospheres absolute (ATA) on neuromuscular endurance performance. We hypothesized that over a 5-day period of consecutive, resting, long-duration hyperoxic WIs there would be a decrease to neuromuscular endurance performance and tissue oxygenation with the quadriceps muscle, but not with the forearm flexors. Thirteen well-trained, male subjects completed five consecutive 6-h resting WIs with 18-h surface intervals during the dive week while breathing 100% oxygen at 1.35 ATA. We assessed skeletal muscle endurance performance before and after each WI, and 24 and 72 h after the final WI. Muscular endurance assessments included 40% maximal handgrip endurance (MHE) and 50-repetition maximal isokinetic (IK) knee extensions. Near-infrared spectroscopy (NIRS) was used to measure muscle oxidative capacity (MOC) of the vastus lateralis and localized muscle tissue oxygenation of the vastus lateralis and flexor carpi radialis. Simultaneously, we measured brachioradialis neuromuscular activation by surface electromyography (SEMG). MHE time-to-fatigue performance declined by 15% at WI 3 (p = 0.009) and by 17% on WI 5 (p = 0.002). Performance continued to decline by 22% at 24-h post-WI (p < 0.001) and by 12% on 72-h post-WI (p = 0.019). Fifty-repetition IK knee extension total work decreased by 5% (p = 0.002) on WI 3, and was further reduced by 7.5 and 12.3% (p = 0.032) at pre-WI 5 and 24-h post-WI, respectively. However, the rate of fatigue was 8 (p = 0.033) and 30% (p = 0.017) lower at WI 3 and 24-h post-WI when compared to WI 1, respectively, demonstrating the muscles were still fatigued from the previous hyperoxic WIs. We detected no significant limitations in oxygen off-loading kinetics during the exercise or MOC measurements. Repeated, resting, long-duration hyperoxic WIs caused significant reductions to muscular endurance but not to indirect measures of oxygen kinetics in load bearing and non-load bearing muscles.


Lateral mass screw fixation in the cervical spine: a systematic literature review.
Lateral mass screw fixation with plates or rods has become the standard method of posterior cervical spine fixation and stabilization for a variety of surgical indications. Despite ubiquitous usage, the safety and efficacy of this technique have not yet been established sufficiently to permit "on-label" U.S. Food and Drug Administration approval for lateral mass screw fixation systems. The purpose of this study was to describe the safety profile and effectiveness of such systems when used in stabilizing the posterior cervical spine. A systematic search was conducted in MEDLINE and the Cochrane Collaboration Library for articles published from January 1, 1980, to December 1, 2011. We included all articles evaluating safety and/or clinical outcomes in adult patients undergoing posterior cervical subaxial fusion utilizing lateral mass instrumentation with plates or rods for degenerative disease (spondylosis), trauma, deformity, inflammatory disease, and revision surgery that satisfied our a priori inclusion and exclusion criteria. Twenty articles (two retrospective comparative studies and eighteen case series) satisfied the inclusion and exclusion criteria and were included. Both of the comparative studies involved comparison of lateral mass screw fixation with wiring and indicated that the risk of complications was comparable between treatments (range, 0% to 7.1% compared with 0% to 6.3%, respectively). In one study, the fusion rate reported in the screw fixation group (100%) was similar to that in the wiring group (97%). Complication risks following lateral mass screw fixation were low across the eighteen case series. Nerve root injury attributed to screw placement occurred in 1.0% (95% confidence interval, 0.3% to 1.6%) of patients. No cases of vertebral artery injury were reported. Instrumentation complications such as screw or rod pullout, screw or plate breakage, and screw loosening occurred in <1% of the screws inserted. Fusion was achieved in 97.0% of patients across nine case series. The risks of complications were low and the fusion rate was high when lateral mass screw fixation was used in patients undergoing posterior cervical subaxial fusion. Nerve root injury attributed to screw placement occurred in only 1% of 1041 patients. No cases of vertebral artery injury were identified in 758 patients. Screw or rod pullout, screw or plate breakage, and screw loosening occurred in <1% of the screws inserted.


Rearrangement of the dendritic morphology of the neurons from prefrontal cortex and hippocampus after subthalamic lesion in Sprague-Dawley rats.
Several studies in rodents have suggested the inactivation of the subthalamic nucleus (STN) as an alternative strategy to Parkinson's disease (PD) treatment. The STN is part of the basal ganglia and plays an important role in the motor function; however, recent data suggest that this structure has a critical role in the cognitive function of the limbic system. The STN receives direct projection from the prefrontal cortex (PFC), structure interconnected with the hippocampus and both structures send excitatory projections to the nucleus accumbens (NAcc). Here, we determined whether and which changes occurred 4 weeks after a STN lesion in the dendritic morphology of pyramidal neurons of the layers 3 and 5 of the PFC and basolateral amygdala, neurons of the ventral hippocampus, and the medium spiny neurons of the NAcc and caudate-putamen. Dendritic morphology was measured using the Golgi-Cox procedure followed by Sholl analysis. We also evaluated the effects of STN lesion on locomotor behavior assessed by an open field test, social interaction, acoustic startle response, prepulse inhibition, and locomotor activity induced by a novel environment and amphetamine. We found that STN damage induced a deficit in locomotion measured by open field test with neuronal hypertrophy in PFC (layer 5) and reduced spinogenesis in CA1 ventral hippocampus and PFC (layer 3). Taken together, these data suggest that the behavioral and morphological effects of STN lesion are, at least partially, mediated by limbic subregions with possible consequences for cognitive-related behaviors observed in PD treatment.


A p53-dependent mechanism underlies macrocytic anemia in a mouse model of human 5q- syndrome.
The identification of the genes associated with chromosomal translocation breakpoints has fundamentally changed understanding of the molecular basis of hematological malignancies. By contrast, the study of chromosomal deletions has been hampered by the large number of genes deleted and the complexity of their analysis. We report the generation of a mouse model for human 5q- syndrome using large-scale chromosomal engineering. Haploinsufficiency of the Cd74-Nid67 interval (containing Rps14, encoding the ribosomal protein S14) caused macrocytic anemia, prominent erythroid dysplasia and monolobulated megakaryocytes in the bone marrow. These effects were associated with defective bone marrow progenitor development, the appearance of bone marrow cells expressing high amounts of the tumor suppressor p53 and increased bone marrow cell apoptosis. Notably, intercrossing with p53-deficient mice completely rescued the progenitor cell defect, restoring common myeloid progenitor and megakaryocytic-erythroid progenitor, granulocyte-monocyte progenitor and hematopoietic stem cell bone marrow populations. This mouse model suggests that a p53-dependent mechanism underlies the pathophysiology of the 5q- syndrome.


Vascular and hematologic effects of hemorrhage in the chicken.
Chickens have been reported to have a rate of posthemorrhagic fluid mobilization twice that of mammals and lack a phase of shock irreversible to transfusion. In the present study we measured several hemodynamic and hematologic parameters in chickens subjected to sustained hemorrhagic hypotension. Total peripheral resistance was unaffected or fell slightly and skeletal muscle vascular resistance judged from changes in gastrocnemius resistance was not affected by hemorrhage. Blood compositional changes included a progressive hyperkalemia, hyperglycemia, and hemodilution as evidenced by linear falls in hematocrit, hemoglobin, and plasma total protein concentration. Plasma sodium and osmolality were unchanged, as were arterial pH and oxygen tension; however, there was a fall in carbon dioxide tension. These studies demonstrate that fluid mobilization in the chicken after hemorrhage does not require an increase in precapillary resistance and suggest that, because the chicken does not exhibit intense precapillary constriction, it is spared some of the deleterious effects of inadequate tissue perfusion. Thus, the findings tend to incriminate the peripheral action of the sympathetic nervous system as a major contributor to the development of irreversibility in other species.


Citicoline in severe traumatic brain injury: indications for improved outcome : A retrospective matched pair analysis from 14 Austrian trauma centers.
Goal-oriented management of traumatic brain injury (TBI) can save the lives and/or improve the long-term outcome of millions of affected patients worldwide. Additionally, enhancing quality of life will save enormous socio-economic costs; however, promising TBI treatment strategies with neuroprotective agents, such as citicoline (CDP-choline), lacked evidence or produced contradictory results in clinical trials. During a prehospital TBI project to optimize early TBI care within 14 Austrian trauma centers, data on 778 TBI patients were prospectively collected. As preceding evaluations suggested a beneficial outcome in TBI patients treated at the Wiener Neustadt Hospital (WNH), we aimed to investigate the potential role of citicoline administration, solely applied in WNH, in those patients. In a retrospective subgroup analysis we compared 67 patients from WNH with citicoline administration and 67 matched patients from other Austrian centers without citicoline use. Patients with Glasgow Coma Scale score <13 on site and/or Abbreviated Injury Scale of the region "head" >2 were included. Our analysis revealed significantly reduced rates of intensive care unit (ICU) mortality (5% vs. 24%, p < 0.01), in-hospital mortality (9% vs. 24%, p = 0.035) and 6‑month mortality (13% vs. 28%, p = 0.031), as well as of unfavorable outcome (34% vs. 57%, p = 0.015) and observed vs. expected ratio for mortality (0.42 vs. 0.84) in the WNH (citicoline receivers) group. Despite the limitations of a retrospective subgroup analysis our findings suggest a possible correlation between early and consequent citicoline administration and beneficial outcomes. Therefore, we aim to set up an initiative for a prospective, multicenter randomized controlled trial with citicoline in sTBI (severe TBI) patients.


Recombinant Zoster Vaccine Significantly Reduces the Impact on Quality of Life Caused by Herpes Zoster in Adult Autologous Hematopoietic Stem Cell Transplant Recipients: A Randomized Placebo-Controlled Trial (ZOE-HSCT).
Herpes zoster (HZ) can have a substantial impact on quality of life (QoL). The vaccine efficacy (VE) of a recombinant zoster vaccine (RZV) was 68.2% (95% confidence interval [CI], 55.6% to 77.5%) in a phase 3 study in adult autologous hematopoietic stem cell transplant (HSCT) recipients (NCT01610414). Herein, we report the impact of RZV on patients' QoL. Autologous HSCT recipients were randomized 1:1 to receive 2 doses of RZV or placebo, given 1 to 2 months apart. QoL was measured by the Short Form Survey-36 and Euro-QoL-5 Dimension at baseline, 1 month, and 1 year postdose 2 and during suspected HZ episodes with the Zoster Brief Pain Inventory (ZBPI). The RZV impact on ZBPI burden of illness and burden of interference scores was estimated. The 2 scores were calculated from the area under the curve (days 0 to 182) of the ZBPI worst pain and ZBPI activities of daily living scores, respectively, assuming a score of 0 for patients not having a confirmed HZ episode. The ZBPI maximum worst pain score was significantly lower in the RZV than placebo group (mean: 5.8 versus 7.1, P = .011). Consequently, the VE estimates for HZ burden of illness (82.5%; 95% CI, 73.6 to 91.4) and burden of interference (82.8%; 95% CI, 73.3 to 92.3) were higher than the HZ VE estimate (ie, 68.2%). RZV showed significantly better QoL scores than placebo 1 week following rash onset among patients with confirmed HZ. In addition to reducing the risk of HZ and its complications, RZV significantly reduced the impact of HZ on patients' QoL in those who developed breakthrough disease.


Evidential support for garlic supplements against diabetic kidney disease: a preclinical meta-analysis and systematic review.
Garlic (Allium sativum L.) is a popular spice that is widely used for food and medicinal purposes and has shown potential effects on diabetic kidney disease (DKD). Nevertheless, systematic preclinical studies are still lacking. In this meta-analysis and systematic review, we evaluated the role and potential mechanisms of action of garlic and its derived components in animal models of DKD. We searched eight databases for relevant studies from the establishment of the databases to December 2022 and updated in April 2023 before the completion of this review. A total of 24 trials were included in the meta-analysis. It provided preliminary evidence that supplementing with garlic could improve the indicators of renal function (BUN, Scr, 24 h urine volume, proteinuria, and KI) and metabolic disorders (BG, insulin, and body weight). Meanwhile, the beneficial effects of garlic and its components in DKD could be related to alleviating oxidative stress, suppressing inflammatory reactions, delaying renal fibrosis, and improving glucose metabolism. Furthermore, time-dose interval analysis exhibited relatively greater effectiveness when garlic products were supplied at doses of 500 mg kg-1 with interventions lasting 8-10 weeks, and garlic components were administered at doses of 45-150 mg kg-1 with interventions lasting 4-10 weeks. This meta-analysis and systematic review highlights for the first time the therapeutic potential of garlic supplementation in animal models of DKD and offers a more thorough evaluation of its effects and mechanisms to establish an evidence-based basis for designing future clinical trials.


A cadaveric study on the stability of lumbar segment after partial laminotomy and facetectomy with intact posterior ligaments.
This study aimed to biomechanically investigate a change in the stability of decompressed segments in the degenerative lumbar spine. Cyclic loading tests to the cadaveric spinal unit were conducted in compressive and bending directions with a stepwise resection of the posterior elements such as wide fenestration (partial laminotomy), unilateral and bilateral facetectomy with the intact supra- and interspinous ligaments and spinous process. Flexion stability did not decrease significantly, and may reflect the fact that the aforementioned ligamentous structures were preserved throughout the resection process. After wide fenestration, the loss of compression stiffness amounted to only half that of bilateral facetectomy, and extension stiffness dropped to this same level. Lateral bending stiffness was maintained until unilateral facetectomy was performed. We recommend restricted decompression surgeries for a single-level central stenosis in the degenerative lumbar spine.


Relationship between K-ras oncogene activation and smoking in adenocarcinoma of the human lung.
To investigate a possible relationship between the exposure to tobacco smoke and the presence of ras point mutations, we examined lung adenocarcinoma samples from 27 smokers and from 27 nonsmokers. Activating point mutations in K-ras (also known as KRAS2) and N-ras (also known as NRAS) were determined by using the polymerase chain reaction and oligonucleotide hybridization to detect the mutated sequences. Mutations were more often found in adenocarcinomas obtained from smokers (eight of 27) than in adenocarcinomas obtained from nonsmokers (two of 27) (P = .044, Fisher's exact test). All mutations were present in K-ras codon 12. None of the other parameters examined differed significantly between the ras-positive and ras-negative groups. We conclude that exposure to carcinogenic agents in tobacco smoke is an important factor in the induction of point mutations in K-ras in human lung adenocarcinomas, but that K-ras mutations may also infrequently occur in tumors of non-smokers.


Effect of cyclo(Leu-Gly) on cyclic GMP-phosphodiesterase activity changes associated with development of tolerance to morphine-induced antinociception, catalepsy, respiratory depression and mydriasis.
A biochemical basis for the development of tolerance to morphine has yet to be defined. Although a number of models have been proposed, none can account for complete tolerance to this drug. Previous studies in our laboratory indicated that the development of complete tolerance to certain morphine-induced behaviors (antinociception, catalepsy and respiratory depression) is associated with changes in the activity of some form(s) of phosphodiesterase with cyclic GMP as substrate (cGMP-PDE) activity in the brain areas that mediate these behaviors (periaqueductal gray, striatum and medulla). In the present study, experiments were performed in which Cyclo(Leu-Gly), a dipeptide that inhibits the development of tolerance to morphine, was administered daily (2 mg/kg) to morphine-naive rats, coadministered with morphine or coadministered with morphine to morphine-tolerant rats and the cGMP-PDE activity was measured. The development of tolerance to the effects was inhibited or reversed by administration of cyclo(Leu-Gly) and there were corresponding changes in cGMP-PDE activity in various brain regions. Differences in cGMP hydrolysis between brain regions from morphine-tolerant animals, tolerance-inhibited animals and tolerance-reversed animals strengthens the evidence for direct involvement of cGMP-PDE(s) in tolerance phenomena.


LuxUS: DNA methylation analysis using generalized linear mixed model with spatial correlation.
DNA methylation is an important epigenetic modification, which has multiple functions. DNA methylation and its connections to diseases have been extensively studied in recent years. It is known that DNA methylation levels of neighboring cytosines are correlated and that differential DNA methylation typically occurs rather as regions instead of individual cytosine level. We have developed a generalized linear mixed model, LuxUS, that makes use of the correlation between neighboring cytosines to facilitate analysis of differential methylation. LuxUS implements a likelihood model for bisulfite sequencing data that accounts for experimental variation in underlying biochemistry. LuxUS can model both binary and continuous covariates, and mixed model formulation enables including replicate and cytosine random effects. Spatial correlation is included to the model through a cytosine random effect correlation structure. We show with simulation experiments that using the spatial correlation, we gain more power to the statistical testing of differential DNA methylation. Results with real bisulfite sequencing dataset show that LuxUS is able to detect biologically significant differentially methylated cytosines. The tool is available at https://github.com/hallav/LuxUS. Supplementary data are available at Bioinformatics online.


Lower serum magnesium levels are associated with more rapid decline of renal function in patients with diabetes mellitus type 2.
Hypomagnesemia has been implicated in adversely affecting diabetic complications. This is a retrospective study designed to determine whether there is any association between serum magnesium concentration [Mg2+] and the rate of renal function deterioration, as determined by the slope of serum creatinine reciprocals versus time (1/SCr-vs-t), in patients with diabetes mellitus type 2 (DM2). DM2 patients without known kidney disease seen at Olive View-UCLA Medical Center for any reason during January-March 2001 were included. For each patient, all available data from our electronic database for [Mg2+], hemoglobin A(1C) (HbA(1C), serum creatinine (SCr), lipid profiles, routine urinary analysis, as well as history of hypertension and pharmacy profiles were retrieved. The average of all parameters obtained and linear regression analyses for the slope of 1/SCr-vs-t plot were performed for each patient. Patients were stratified by gender and divided into four groups based on increasing [Mg2+]. Correlations between each parameter including the slope of 1/SCr-vs-t and the four magnesium groups were analyzed. 252 males and 298 females with a mean follow-up of 62.6 +/- 22.5 months were included. Patients belonging to lower [Mg2+] groups for both genders had significantly worse slopes of 1/SCr-vs-t plot independent of the presence of hypertension and use of ACEI/ARB, diuretics, HMG-CoA enzyme inhibitors or aspirin. In a multivariate regression analysis controlling for age, HbA(1C) and various components of the lipid profile, [Mg2+] remained an independent predictor for the slope of 1/SCr-vs-t. A trend for worse proteinuria based on routine urinary analysis was observed among patients belonging to the lowest [Mg2+] group. Lower [Mg2+] is associated with a faster renal function deterioration rate in DM2 patients.