Isotype-Specific Detection of Serum Immunoglobulins Against Allergens.
Type-I hypersensitivity is commonly characterized by increased levels of antigen-specific immunoglobulin (Ig) E. Therefore, it is important for clinical and research investigators to reliably measure serum levels of IgE in allergic patients and animal models. While current ELISA-based methods are simple and commonly performed for the detection of allergen-specific IgE using serum or plasma, they may produce misleading results. This is in part due to decreased sensitivity for IgE in the presence of other Ig isotypes in the same sample, such as IgG, that are typically more abundant than IgE. When assessment of multiple Ig isotypes is necessary, performing optimized assays for individual isotypes requires high sample volumes. Here, we describe an approach to increase the sensitivity for IgE detection while conserving the sample volume needed. This method not only improves the accuracy of serum IgE measurements but also allows simultaneous analysis of other allergen-specific immunoglobulins.


Home negative pressure ventilation: report of 20 years of experience in patients with neuromuscular disease.
Twenty years of experience using negative pressure devices (NPD) at home to ventilate 40 patients with neuromuscular disease is presented. The purpose of the study was to determine the costs, complications, and clinical outcome of this form of respiratory support, and to ascertain the reasons for failure to institute effective negative pressure ventilation (NPV) in nine patients. Emerson tank respirators, used mainly to rest respiratory muscles at night, and intermittent positive pressure breathing machines were used by 98% of patients at an average equipment cost of +2,700 annually. Patients in whom NPV was initiated on an elective rather than emergent basis saved an average of +12,000 during their initial hospitalization. Life table analysis shows a five-year survival of 76%, and a 10-year survival of 61%. Complications were minor and occurred at an average rate of less than one per year per patient at home on NPV. Failure to achieve satisfactory NPV in nine patients was associated with age (six patients were younger than 3 years of age), or severe thoracocervical scoliosis, which prevented proper fitting of the NPD. For reasons of safety, economy, and quality of life, NPV at home is the preferred treatment for patients having neuromuscular disease who need respiratory assistance.


Structural MRI in normal aging and Alzheimer's disease: white and black spots.
White matter changes (WMC) and microbleeds (MBs) are common in the elderly and in patients with Alzheimer's disease (AD). To describe the prevalence, anatomical distribution and longitudinal changes of WMC and MBs in normal aging subjects compared to AD patients and to describe current evidence of their effects on the clinical course of AD. Short literature review. WMC and MBs are more frequent and progress more rapidly in AD patients than in cognitively normal elderly. New MBs occur in up to one quarter of AD cases without any therapeutic intervention. WMC and MBs influence the clinical course of AD. WMC and MBs might represent treatment targets in clinical trials on AD.


Factors that affect the sensitivity of the in vivo-in vitro hepatocyte DNA repair assay in the male rat.
Measurement of chemically induced DNA repair as unscheduled DNA synthesis (UDS) in rat hepatocytes following in vivo exposure has been shown to be a useful indicator of the genotoxicity of chemicals in rat liver. We have examined some of the parameters of this assay in an attempt to increase its sensitivity and reduce cytoplasmic backgrounds. Fischer-344 rats were treated with a low dose of a known positive chemical, water, or corn oil. Livers were perfused with a collagenase solution and isolated hepatocytes were incubated with [3H]thymidine (3H-TdR) followed by overnight incubation in unlabeled TdR, then cell fixation and washing. UDS was measured by quantitative autoradiographic grain-counting as net grains/nucleus (NG). Incubation in 3H-TdR ranging in age from 1 week to more than 12 months gave highly variable background (BKG) and NG counts and a slight overall decrease in NG when the 3H-TdR used was more than 4 months old. Control BKG was 3 times higher after 19 h than after 4 h of incubation in 3H-TdR, with little change in NG. Incubation in unlabeled TdR also reduced BKG significantly. Reduction of autoradiogram exposure from two to one week cut BKG in half without significantly reducing NG. A half-hour wash in fixative (1:3 acetic acid:ethanol) followed by two water washes was as effective in reducing BKG as three 10-min washes in fixative followed by 6 water washes, and resulted in better overall cell attachment. An examination of the distribution of historical control data shows that vehicle control animals never exceed zero NG. This suggests that any NG response greater than zero should be viewed as a possible positive response.


Effects of COVID-19 on heart failure admissions.
Despite significant investigation into the effects of COVID-19 on cardiovascular disease, there is a paucity of national data specifically examining its effects on heart failure (HF) hospitalizations. Previous cohort study data demonstrate worsened outcomes in HF patients with recent COVID-19 infection. To better understand this association, this study aimed to utilize a nationally representative database to examine demographics, outcomes, and health care utilization in hospitalizations for HF with a codiagnosis of COVID-19.


The concentrations of copper and zinc in human milk. A longitudinal study.
Twenty-seven healthy Finnish mothers were followed during the course of their entire lactation period. A total of 229 individual milk samples, collected in the beginning and at the end of each feed during a 24-h period, were obtained from the 2nd week to the 9th month of lactation. The copper and zinc concentrations were determined by atomic absorption spectrophotometry. The concentrations of the trace-elements investigated were dependent on the stage of lactation. The median copper and zinc concentrations decreased during the course of lactation from about 0.60 mg/l and 4.0 mg/l to 0.25 mg/l and 0.5 mg/l, respectively. The importance of considering the stage of lactation in the evaluation of the trace-element nutrition value of breast milk should be emphasized. The calculated means of the concentrations of these trace-elements in mature human milk presented in the literature seem to overestimate the actual levels in prolonged lactation.


Inhibition of apoptosis by miR‑122‑5p in α‑fetoprotein‑producing gastric cancer.
α‑Fetoprotein (AFP)‑producing gastric cancer (AFPGC) is recognized as one of the most aggressive tumors with subsequent poor prognosis compared with common gastric cancer (GC) subtypes. However, the molecular mechanism remains to be elucidated. We previously identified that miR‑122‑5p could be a useful biomarker in AFPGC patients. We examined herein the biological function of miR‑122‑5p and the molecular mechanism underlying tumor progression in AFPGC. We used the AFPGC cell line (FU97) and miR‑122‑5p inhibitor to examine the function of miR‑122‑5p. Moreover, we investigated the possible targets of miR‑122‑5p. The expression level of miR‑122‑5p was significantly increased in the FU97 cell line than in common GC cell lines. Also, suppression of miR‑122‑5p significantly reduced AFP levels and proliferation in AFPGC through an induction of apoptosis. Western blotting revealed that the expression of anti‑apoptotic protein (Bcl‑2) was decreased and that of pro‑apoptotic protein (caspase‑3) was increased in miR‑122‑5p suppression of FU97. Moreover, we revealed that FOXO3 was an important target of miR‑122‑5p in AFPGC, which inhibited apoptosis and subsequently manifested aggressiveness. In conclusion, miR‑122‑5p inhibited apoptosis and facilitated tumor progression by targeting FOXO3 in AFPGC, which indicates the possibility of miR‑122‑5p as a potential therapeutic target in AFPGC.


Muscle and plasma amino acids during semi-starvation in normal subjects: hypocaloric glucose vs. amino acid infusions.
Normal subjects were fed hypocaloric amounts of amino acids or glucose for 3 days to separate the effects of these nutrients from those of trauma on muscle and plasma amino acids. Intracellular and plasma levels of Val, Leu and Ile were increased in both groups but were increased to a greater extent when amino acids were infused. Intracellular free glutamine concentration in muscle tissue was not significantly altered. Some of the trauma induced changes in amino acids can be replicated by maintaining normal subjects on hypocaloric amino acids; the decrease in glutamine concentration seen with trauma is not a function of hypocaloric nutrition.


Guessing as a learning intervention: A meta-analytic review of the prequestion effect.
Giving students test questions before they have learned the correct answers (i.e., prequestions) enhances learning. However, existing research has provided conflicting evidence on whether the benefits of prequestions are specific to the initially tested material or if they generalize to new, nontested material. In this review, we summarize the literature on the prequestion effect, describe the attention-based account underlying this effect, report a meta-analysis of the magnitude of the specific and general effects, and explore theoretically and empirically relevant moderator variables that influence the size and direction of the prequestion effect. This preregistered meta-analysis demonstrated a moderate specific effect (g = 0.54, k = 97) but a virtually nonexistent general effect (g = 0.04, k = 91). Overall, the attention-based account received support from some theoretically relevant moderator analyses. Future researchers are encouraged to conduct theoretically motivated studies to help clarify the mechanisms that underlie the attention-enhancing effects of prequestions and to explore the benefits of prequestions in educational domains to establish the extent to which these effects translate into the classroom.


Guidelines and Initiatives for Good Research Practice.
This chapter explores existing data reproducibility and robustness initiatives from a cross-section of large funding organizations, granting agencies, policy makers, journals, and publishers with the goal of understanding areas of overlap and potential gaps in recommendations and requirements. Indeed, vigorous stakeholder efforts to identify and address irreproducibility have resulted in the development of a multitude of guidelines but with little harmonization. This likely results in confusion for the scientific community and may pose a barrier to strengthening quality standards instead of being used as a resource that can be meaningfully implemented. Guidelines are also often framed by funding bodies and publishers as recommendations instead of requirements in order to accommodate scientific freedom, creativity, and innovation. However, without enforcement, this may contribute to uneven implementation. The text concludes with an analysis to provide recommendations for future guidelines and policies to enhance reproducibility and to align on a consistent strategy moving forward.


Association between low fetal fraction in cell-free DNA screening and fetal chromosomal aberrations: A systematic review and meta-analysis.
To perform a systematic review and meta-analysis of the available literature on low fetal fraction (LFF) in cell-free DNA (cfDNA) screening and the risk of fetal chromosomal aberrations. We searched articles published between January 2010 and May 2021 in PubMed and EMBASE databases. Risk of bias was assessed using QUADAS-2. Twenty-seven studies met the inclusion criteria, comprising data of 243,700 singleton pregnancies. Compared to normal fetal fraction, LFF was associated with a higher risk of trisomy 13 (OR 5.99 [3.61-9.95], I 2 of heterogeneity = 0%, n = 22 studies), trisomy 18 (OR 4.46 [3.07-6.47], I 2 = 0%, n = 22 studies), monosomy X (OR 5.88 [2.34-14.78], I 2 = 18%, n = 10 studies), and triploidy (OR 36.39 [9.83-134.68], I 2 = 61%, n = 6 studies), but not trisomy 21 (OR 1.25 [0.76-2.03], I 2 = 36%, n = 23 studies). LFF was also associated with a higher risk of various other types of fetal chromosomal aberrations (OR 4.00 [1.78-9.00], I 2 = 2%, n = 11 studies). Meta-analysis of proportions showed that absolute rates of fetal chromosomal aberrations ranged between 1% and 2% in women with LFF. A limitation of this review is the potential risk of ascertainment bias because of differences in outcome assessment between pregnancies with LFF and those with normal fetal fraction. Heterogeneity in population characteristics or applied technologies across included studies may not have been fully addressed. An LFF test result in cfDNA screening is associated with an increased risk of fetal trisomy 13, trisomy 18, monosomy X, and triploidy, but not trisomy 21. Further research is needed to assess the association between LFF and other specific types of fetal chromosomal aberrations.


Mechanistic basis for unexpected bioavailability enhancement of polyelectrolyte complexes incorporating BCS class III drugs and carrageenans.
The objective of this study was to investigate the potential of λ-carrageenan to work as an absorption modifying excipient in combination with formulations of BCS class 3 substances. Trospium chloride was used as a model BCS class 3 substance. Polyelectrolyte complexes of trospium and λ-carrageenan were produced by layer-by-layer complexation. A λ-carrageenan-containing formulation was administered either in capsules size 9 to rats by gavage or directly into ligated intestinal loops of rats. Exceptionally strong variations were observed in the plasma concentrations of the rats that received λ-carrageenan compared to the control group, but enhanced plasma concentrations were observed only in some of the rats. In vitro permeability studies were performed across Caco2-monolayers and across excised segments of rat jejunum in a modified Ussing chamber to learn more about the mechanism of absorption enhancement. The complex did not show any effect in Caco2-cells, but led to a major enhancement of permeability across excised segments in modified Ussing chambers. Carrageenan did not lead to alterations of tight junctions. The bioavailability enhancing effect thus was most likely due to an interaction of the polyelectrolyte-drug complex with the mucus, which provided an intimate contact between the drug and the absorbing surface. A similar effect was also achievable with other types of carrageenan and was also transferable to other compounds. In conclusion, λ-carrageenan-drug complexes show interesting excipient-drug-epithelium interactions - however, for full utilization of the permeation enhancing potential, an intimate and reproducible contact between absorbing epithelia and the complex is needed.


Glycoconjugates secreted by bovine tracheal serous cells in culture.
Glycoconjugates secreted by bovine tracheal gland serous cells in culture were characterized after incorporation of radioactive precursor [1-14C]glucosamine and stimulation with isoproterenol. Under dissociative conditions, glycoconjugates eluted in both the void and included volumes on Sepharose Cl-4B. Fractionated by anion-exchange chromatography, the high-molecular-weight (Sepharose Cl-4B; V0) glycoconjugates gave two acidic fractions eluting at 0.5 and 2.0 M NaCl; low-molecular-weight glycoconjugates of the included volumes gave a neutral fraction and two acidic fractions eluting at 0.5 and 2.0 M NaCl. Based on chemical analysis and specific enzymatic digestions, the material eluting in the void volume was shown to contain hyaluronic acid and chondroitin sulfate proteoglycan. In addition, the presence of small amounts of galactose, fucose, sialic acid, glucosamine, and galactosamine suggest the presence of O-glycosidically linked glycoproteins in the void volume. The identification of galactosaminitol in beta-eliminated oligosaccharides from this material confirms this notion. The material eluting in the included volume was shown to contain N-linked glycoproteins with glycans of complex type in the neutral fraction and chondroitin sulfate proteoglycans in the two acidic fractions. Significant N-sulfation of amino sugars was detected in the 0.5 M acidic fraction, indicating the presence of heparan sulfate. Hyaluronic acid and chondroitin sulfate proteoglycan have recently been identified in tracheal secretions; our results suggest that these components originate at least in part from tracheal gland serous cells.


Novel G protein-coupled receptors: a gene family of putative human olfactory receptor sequences.
We have taken advantage of the sequence conservation in the G protein-coupled receptor superfamily to isolate a fragment of a novel G protein-coupled receptor sequence using polymerase chain reaction (PCR) amplification of human genomic DNA. Screening of human genomic and hippocampal cDNA libraries with this amplified receptor fragment revealed a number of related sequences. Sequence analysis of four genomic clones and one cDNA clone clearly identifies these as related members of the G protein-coupled receptor family, as the deduced amino acid sequence reveals putative transmembrane domains and conserved amino acid residues. Southern blot analysis of restriction digests of human genomic DNA indicates that these receptor subtypes are likely to belong to a family of related genes. One of the proposed receptor sequences indicates the presence of pseudogenes in this family. Based on the homology of these sequences to a family of recently described receptors expressed exclusively in rat olfactory epithelium, it is suggested that these receptors represent a family of human odorant receptors.


Management of treatment-resistant depression.
Given the limitations of evidence for treatment options that are consistently effective for TRD and the possibility that TRD is in fact a form of depression that has a low probability of resolving, how can clinicians help patients with TRD? Perhaps the most important conceptual shift that needs to take place before treatment can be helpful is to accept TRD as a chronic illness, an illness similar to many others, one that can be effectively managed but that is not, at our present level of knowledge, likely to be cured. An undue focus on remission or even a 50% diminution of symptoms sets unrealistic goals for both patients and therapists and may lead to overtreatment and demoralization. The focus should be less on eliminating depressive symptoms and more on making sense of and learning to function better in spite of them. It is important to acknowledge the difficult nature of the depressive illness, to remove blame from the patient and clinician for not achieving remission, to set realistic expectations, and to help promote better psychosocial functioning even in the face of persisting symptoms. The critical element when implementing such an approach is a judicious balance between maintaining hope for improvement without setting unrealistic expectations. It is important to reemphasize that following a disease management model with acceptance of the reality of a chronic illness is not nihilistic and does not mean the abandonment of hope for improvement. The first step in treating a patient with TRD is to perform a comprehensive assessment of the patient's past and current treatment history to ensure that evidence-based treatment trials have in fact been undertaken, and if not, such treatment trials should be implemented. If the patient continues to have significant residual symptoms, it is important to determine the impact is of these symptoms on the patient's quality of life and ability to function. It is also important to evaluate the factors that may be contributing to the persistence of depressive symptoms such as comorbid personality disorders, somatic disorders, substance abuse, and work and interpersonal conflicts. The treatment of patients with TRD needs to move beyond attempts to modify symptoms without taking into consideration and attempting to modify the patient's personality, coping skills, and social system. Further somatic treatment trials can be undertaken, if desired by the patient and therapist, as a small (5%–15%) percentage of patients may respond and further treatment trials, and this may engender hope. The risk with this approach is that patients and therapists may not work at disease management skills if they believe there may be a resolution of the depression if they could just find the right medication or intervention. Therapists may also feel pressured by patients, families, insurance companies, as well as their own sense of helplessness to escalate treatment in a more and more aggressive manner in an attempt to achieve an elusive remission. A disease management program can provide the therapist and patient with sufficient structure, skills, and goals to encourage ongoing treatment without resorting to unproven measures that may create more side effects and problems. It is particularly important to include the patient's significant others in the reformulation of the patient's problem and thereby learn how to manage the illness more effectively. Significant others and family members can be invaluable in providing support for dealing with the difficult process of acquiring a new skill set. Indeed, they spend significantly more time with the patient than does any therapist. Family members are likely to provide this kind of support only if they have been part of the assessment and treatment process. Patients with a wide range of chronic medical illnesses can and do learn to function effectively and to achieve a satisfying quality of life in spite of their illness. There is no reason to think that patients with TRD should not be able to achieve a similar level of illness management, functioning, and quality of life.


In vivo prediction of abdominal fat and breast muscle in broiler chicken using live body measurements based on machine learning.
The purpose of this study was to predict the carcass characteristics of broilers using support vector regression (SVR) and artificial neural network (ANN) model methods. Data were obtained from 176 yellow feather broilers aged 100-day-old (90 males and 86 females). The input variables were live body measurements, including external measurements and B-ultrasound measurements. The predictors of the model were the weight of abdominal fat and breast muscle in male and female broilers, respectively. After descriptive statistics and correlation analysis, the datasets were randomly divided into train set and test set according to the ratio of 7:3 to establish the model. The results of this study demonstrated that it is feasible to use machine learning methods to predict carcass characteristics of broilers based on live body measurements. Compared with the ANN method, the SVR method achieved better prediction results, for predicting breast muscle (male: R2 = 0.950; female: R2 = 0.955) and abdominal fat (male: R2 = 0.802; female: R2 = 0.944) in the test set. Consequently, the SVR method can be considered to predict breast muscle and abdominal fat of broiler chickens, except for abdominal fat in male broilers. However, further revaluation of the SVR method is suggested.


The use of the abdominal circumference as a means of assessing M-mode ventricular dimensions during the second and third trimesters of pregnancy in the normal human fetus.
Eighty-two normal fetuses were scanned from 18 to 41 weeks of gestation. The following M-mode measurements were correlated with the abdominal circumference: diastolic biventricular outer (r = 0.951) and inner (r = 0.948) dimensions; diastolic right (r = 0.942) and left (r = 0.925) internal dimensions; tricuspid (r = 0.916) and mitral (r = 0.900) valve opening excursions; and right (r = 0.774), left (r = 0.731), and interventricular septal (r = 0.753) wall thickness. Polynomial regression demonstrated that linear regression best described the data from which the mean and 5 per cent and 95 per cent confidence limits for predicted M-mode measurements were computed for the abdominal circumference between 12 and 35 cm.


Antimicrobial activity of nerolidol and its derivatives against airborne microbes and further biological activities.
Nerolidol and its derivatives, namely cis-nerolidol, O-methyl-nerolidol, O-ethyl-nerolidol, (-)-α-bisabolol, trans,trans-farnesol and its main natural source cabreuva essential oil, were tested for their antimicrobial activity against airborne microbes and antifungal properties against plant pathogens. Among the tested compounds, α-bisabolol was the most effective antimicrobial agent and trans,trans-farnesol showed the best antifungal activity.


Replacement analysis of non-equilibrium competition between rock pool Corixids (Hemiptera, Corixidae).
The resource use of rock pool Corixids Arctocorisa carinata (Sahlb.) and Callicorixa producta (Reut.) was compared in single-species and mixed populations established in the natural environment. The experimental recruitment consisted of daily transfer of newly moulted first-stage larvae to the study pools. The changes in numbers of larvae and the increase of the biomass corresponded closely to the changes in natural populations. The growth of populations was limited by elimination of the youngest individuals. The maximal biomasses attained per unit of pool surface area were same in all cases. This indicates that the species have completely overlapping resources and use them with nearly equal efficiency. The strong interference by A. carinata is an additional reason for the unstability of the competitive interaction.


Azido Homoalanine is a Useful Infrared Probe for Monitoring Local Electrostatistics and Sidechain Solvation in Proteins.
The use of IR probes to monitor protein structure, deduce local electric field, and investigate the mechanism of enzyme catalysis and protein folding has attracted increasing attention. Here, the azidohomoalanine (Aha) is considered as a useful IR probe. The intricate details of the distinct effects of backbone peptide bonds and H-bonded water molecules on the azido stretch mode of the IR probe Aha were revealed by carrying out QM/MM MD simulations of two variants of the protein NTL9, NTL9-Met1Aha and NTL9-Ile4Aha and comparing the resulting simulated IR spectra with experiments.


Bacterial signaling as an antimicrobial target.
Antibiotics profoundly reduced worldwide mortality. However, the emergence of resistance to the growth inhibiting effects of these drugs occurred. New approaches to treat infectious disease that reduce the likelihood for resistance are needed. In bacterial pathogens, complex signaling networks regulate virulence. Anti-virulence therapies aim to disrupt these networks to attenuate virulence without affecting growth. Quorum-sensing, a cell-to-cell communication system, represents an attractive anti-virulence target because it often activates virulence. The challenge is to identify druggable targets that inhibit virulence, while also minimizing the likelihood of mutations promoting resistance. Moreover, given the ubiquity of quorum-sensing systems in commensals, any potential effects of anti-virulence therapies on microbiome function should also be considered. Here we highlight the efficacy and drawbacks of anti-virulence approaches.


Absence of mechanical hyperalgesia after exercise (delayed onset muscle soreness) in neonatally capsaicin-treated rats.
Delayed onset muscle soreness (DOMS) appears with some delay after unaccustomed, strenuous exercise, especially after lengthening contraction (LC). It is characterized by tenderness and movement related pain, namely muscular mechanical hyperalgesia. To clarify the involvement of C-fibers in this mechanical hyperalgesia, we examined whether DOMS could be induced in rats treated neonatally with capsaicin. We confirmed that a large portion of unmyelinated afferent fibers were lost in capsaicin treated rats. In these animals, LC failed to induce muscular mechanical hyperalgesia. mRNA of nerve growth factor (NGF) in the muscle, which plays a pivotal role in maintaining mechanical hyperalgesia, was upregulated in the capsaicin treated animals similar to the vehicle treated animals. These results demonstrate that C-fiber afferents are essential in transmitting the nociceptive information from exercised muscle in DOMS.


[Aspartate-carbamoyltransferase activity of rabbit liver tissue following x-irradiation and vitamin and ATP administration].
Under the effect of total X-ray irradiation (450 R) the changes in activity of aspartate-carbamoyltransferase in the rabbit liver are of wavy character depending on the time passed after irradiation. Administration of vitamin B (with ATP, in combination with ATP and B1 as well as with FMN and PP) is shown to produce an activating effect on the enzyme function in the irradiated animals.


Dissecting spindle architecture with a laser.
Microtubules in spindles are too dense to resolve by light microscopy, even with super-resolution methods. Using a new method based on laser-ablation techniques, Brugués et al. present the first quantitative characterization of the vertebrate meiotic spindle and propose an assembly mechanism for building this architecture.


LEF-1: Diagnostic utility in distinguishing basaloid neoplasms of the salivary gland.
Lymphoid enhancer binding factor 1 (LEF-1) has recently been reported as a potential immunohistochemical (IHC) marker for basal cell adenoma (BCA) and other salivary gland tumors, which may contribute to an increased accuracy in differentiating basaloid salivary gland neoplasms. We evaluated the utility of LEF-1 in fine needle aspiration (FNA) and resection specimens to distinguish pleomorphic adenoma (PA), BCA, basal cell adenocarcinoma (BCAC), and adenoid cystic carcinoma (ACC) as well as in non-neoplastic salivary gland (NNSG). Cases including 66 PA (35 FNA, 31 resections), 12 BCA (5 FNA, 7 resections), 42 ACC (11 FNA, 31 resections), 1 BCAC FNA, and 10 NNSG (5 FNA, 5 resections) were obtained and stained for LEF-1. On cell block (CB), 51% of PA and 60% of BCA were LEF-1 positive while 91% of ACC were LEF-1 negative. Among resections, there was a higher percentage of LEF-1 positive PA (84%) and BCA (86%), and a higher percentage of LEF-1 negative ACC (97%). LEF-1 staining had a low to moderate sensitivity for detecting benign basaloid neoplasms on FNA CB and resection specimens (52.5% and 84%, respectively), but a higher specificity (92% and 97% respectively), and positive predictive value (95% and 97% respectively). When comparing benign (PA and BCA) and the most common malignant basaloid salivary gland tumor (ACC), positive LEF-1 favors a benign neoplasm. Additional studies with LEF-1, specifically including other rare basaloid salivary gland neoplasms are needed to further clarify the role of LEF-1 in diagnosing these lesions on FNA.


Delirium in the elderly: a review.
Delirium is a common disorder, often under diagnosed and mismanaged. It is becoming more prevalent, because of the ageing population. In this clinical review, we summarise the definition, diagnosis and management of delirium.


Epilepsy with myoclonic-atonic seizures (Doose syndrome): When video-EEG polygraphy holds the key to syndrome diagnosis.
An electroclinical epilepsy syndrome diagnosis enables physicians to predict outcomes as well as select appropriate treatment options. We report a child who presented with reflex myoclonus at the age of 9 months and was initially diagnosed with myoclonic epilepsy in infancy. After 9 years of medically resistant myoclonic seizures, extensive investigations, and emerging learning difficulties, she was referred for video-telemetry to characterize her seizures in an attempt to make a syndromic diagnosis. A three-day video-telemetry assessment was performed to document seizures. Neck and deltoid EMG channels were applied from the onset of the recording. Frequent generalized bursts of 3- to 5-Hz spike/polyspike and slow wave discharges, associated with clinical manifestations, mostly myoclonic seizures, were noted. In addition, definite atonic components were noted on the neck EMG as well as the deltoids associated with the slow component of the ictal discharges. The EEG and polygraphy findings are suggestive of a generalized epilepsy characterized by predominantly myoclonic seizures with atonic components. This raises the possibility whether a variant of epilepsy with myoclonic-atonic seizures (Doose syndrome) may be the underlying diagnosis for this girl. A trial of the ketogenic diet would therefore be considered as an option in her future management in view of its beneficial effect in this condition.


The sorLA cytoplasmic domain interacts with GGA1 and -2 and defines minimum requirements for GGA binding.
We report that the Vps10p domain receptor sorLA binds the adaptor proteins GGA1 and -2, which take part in Golgi-endosome sorting. The GGAs bind with differential requirements via three critical residues in the C-terminal segment of the sorLA cytoplasmic tail. Unlike in sortilin and the mannose 6-phosphate receptors, the GGA-binding segment in sorLA contains neither an acidic cluster nor a dileucine. Our results support the concept of sorLA as a potential sorting receptor and suggest that key residues in sorLA and sortilin conform to a new type of motif (psi-psi-X-X-phi) defining minimum requirements for GGA binding to cytoplasmic receptor domains.


New insights into enhanced biodegradation of 4-bromphenol in a nitrate-reducing system: Process performance and mechanism.
Due to the recalcitrant nature of halogenated phenol, conventional anaerobic bioprocess is often limited by low removal efficiency and poor process stability. At the presence of electron acceptors such as nitrate, 4-bromophenol (4-BP) removal efficiency is significantly higher than that in the anaerobic control system, but the mechanism involved is still unclear. Therefore, an up-flow nitrate-reducing bioreactor (NRBR) was designed and consecutively performed for 215 days to explore the synergistic mechanism for BPs biodegradation and nitrate reduction. Complete 4-BP biodegradation could be obtained in NRBR at HRT and 4-BP loading rate of 24 h and 0.29 mol m - 3d - 1, while the TOC removal and nitrate reduction efficiencies were as high as 91.33±2.11% and 98.31±1.33%, respectively. Population evolution analyses revealed that the microorganisms involved in 4-BP debromination and biodegradation (Candidatus Peregrinibacteria, Denitratisoma, Anaerolineaceae and Ignavibacterium) as well as nitrate reduction (Denitratisoma, Anaerolineaceae, Limnobacter and Ignavibacterium) were significantly enriched in NRBR. Major intermediates during 4-BP biodegradation, including 4-bromocatechol, 4‑bromo-6-oxo-hexanoic acid and succinic acid were identified, while a distinct 4-BP biodegradation pathway via hydration, aromatic-ring cleavage, hydrolysis debromination and oxidation was expounded. Metagenomic analysis indicated that oxidation (had, pht4, boh, butA), hydrolysis debromination ((S)-2-haloacid dehalogenase) and bio-mineralization (gabD, sdhA) of 4-BP were largely enhanced in NRBR. Moreover, carbon, nitrogen, energy and amino acid metabolisms were significantly facilitated with the injection of nitrate in order to provide energy and electron, thus enhanced microbial activities and enzymatic reactions in NRBR. The proposed mechanism provides new insights into our mechanistic understanding of halogenated phenol biodegradation and the development of sustainable bioremediation strategies.


TIGAR contributes to ischemic tolerance induced by cerebral preconditioning through scavenging of reactive oxygen species and inhibition of apoptosis.
Previous study showed that TIGAR (TP53-induced glycolysis and apoptosis regulator) protected ischemic brain injury via enhancing pentose phosphate pathway (PPP) flux and preserving mitochondria function. This study was aimed to study the role of TIGAR in cerebral preconditioning. The ischemic preconditioning (IPC) and isoflurane preconditioning (ISO) models were established in primary cultured cortical neurons and in mice. Both IPC and ISO increased TIGAR expression in cortical neurons. Preconditioning might upregulate TIGAR through SP1 transcription factor. Lentivirus mediated knockdown of TIGAR significantly abolished the ischemic tolerance induced by IPC and ISO. ISO also increased TIGAR in mouse cortex and hippocampus and alleviated subsequent brain ischemia-reperfusion injury, while the ischemic tolerance induced by ISO was eliminated with TIGAR knockdown in mouse brain. ISO increased the production of NADPH and glutathione (GSH), and scavenged reactive oxygen species (ROS), while TIGAR knockdown decreased GSH and NADPH production and increased the level of ROS. Supplementation of ROS scavenger NAC and PPP product NADPH effectively rescue the neuronal injury caused by TIGAR deficiency. Notably, TIGAR knockdown inhibited ISO-induced anti-apoptotic effects in cortical neurons. These results suggest that TIGAR participates in the cerebral preconditioning through reduction of ROS and subsequent cell apoptosis.


Cineroentgenographic and electromyographic studies of Eustachian tube function.
To determine the opening and closing process of the Eustachian tube during swallowing, the simultaneous recording of cineroentgenograms and electromyograms (EMG) was carried out in three monkeys. It was revealed that synchronous with the activity of the tensor veli palatini muscle, the cartilaginous tube opened for a very short time (ca. 250 ms) by an outward movement of its lateral wall. On the other hand, the pharyngeal orifice of the tube dilated for a relatively long time synchronous with the activity of the levator veli palatini muscle. These time lags between two parts of the tube seemed to help the pumping function of the tube.


The ultrastructure of the sinus gland of the crayfish Astacus leptodactylus (Nordmann).
An ultrastructural study of the sinus gland of the crayfish Astacus leptodactylus demonstrates that this gland is mainly composed of glial cells, axons and axon terminals. On the basis of the size, shape and electron density of the neurosecretory granules, we could distinguish five different types of axon terminals.


The performance of modified blood pressure-to-height ratio as a screening measure for identifying children with hypertension.
This study evaluated the accuracy of modified blood pressure-to-height ratio (MBPHR) for identifying hypertension in Han children aged 7-12 years. In 2011, anthropometric measurements were assessed in a cross-sectional population-based study of 1352 Han children aged 7-12 years. Elevated blood pressure was defined according to the 2004 National High Blood Pressure Education Program Working Group definition (as gold standard). The following equations for MBPHR were used: modified systolic blood pressure to height ratio(MSBPHR) = SBP(mmHg)/(height(cm) + 7 × (13 - age(years))), modified diastolic blood pressure to height ratio (MDBPHR) = DBP(mmHg)/(height(cm) + 7 × (13 - age(years))). Receiver operating characteristic curve analyses were performed to assess the accuracy of MSBPHR and MDBPHR as diagnostic tests for elevated SBP and DBP, respectively. The accuracy of MSBPHR and MDBPHR (assessed by area under the curve) for identifying elevated SBP and DBP were over 0.85 (0.953-1.000). When elevated blood pressure was defined by MBPHR (age-dependent cut-off point), the sensitivities were 99.1% in boys and 97.0% in girls and the specificities were 89.0% in boys and 92.3% in girls. When elevated blood pressure was defined by MBPHR (non-age-dependent cut-off point), the sensitivities were 96.4% in boys and 99.2% in girls and the specificities were 81.2% in boys and 75.5% in girls. MBPHR is an accurate index for screening hypertension in children, but is not superior to BPHR. Compared with age-dependent BPHR cutoff points, non-age-dependent MBPHR cut-off point is simple but increase the proportion of reexamination.


Prevalence, behavioural, and management outcomes of infants with auditory neuropathy spectrum disorder.
To provide an overview of electrophysiological and behavioural outcomes from a large UK centres data set on children diagnosed with auditory neuropathy (ANSD) between 2002 and March 2018. A systematic audit was undertaken, collating the electrophysiological data from auditory brainstem response (ABR) follow-up, risk factors, and later behavioural results/management. These were then compared to look for trends between groups. The study sample consisted of 118 925 infants born, with 46 (0.039%, 0.39 per 1000 births) being diagnosed with congenital ANSD (39 bilateral, seven unilateral). Twenty-nine per cent of ears with ANSD had short latency components on ABR testing. Forty-four per cent of ears with present cochlear microphonics but absent transient-evoked otoacoustic emissions (TE-OAE) and no ABR went on to have profound behavioural hearing threshold levels. All but one child went on to show a hearing loss on behavioural testing. ANSD was not confined to the population from neonatal intensive care units: there were three bilateral and five unilateral cases in the typically developing infant population. The incidence of ANSD is higher in this sample than that reported previously in the literature. Children who had cochlear microphonics with absent ABR and absent TE-OAE had significantly worse later behavioural outcomes than other patterns of electrophysiological results.


Probing structural and motional features of organic and inorganic solids through extended family of cross-polarization experiments.
Combined use of cross-polarization and magic-angle spinning in the middle of the seventies has opened a new era of high-resolution solid-state NMR spectroscopy. Cross-polarization procedure is commonly used to obtain a shorter measuring time and to investigate or exploit one nucleous by means of the other nucleous involved in the polarization transfer. An extended family of cross-polarization experiments including constant time cross-polarization approach, cross-polarization inversion and indirect observation of proton spin system is reviewed and illustrated with applications to a large range of solids.


The kinetics of monospermic and polyspermic fertilization in free-spawning marine invertebrates.
The equation of Vogel et al. (1982) is widely used in fertilization studies of free-spawning marine invertebrates to predict the percentage of viable eggs that will be fertilized at any specified levels of gamete concentration and contact time. Here, the random collision model that underlies the Vogel et al. equation is extended to distinguish between monospermic and polyspermic fertilization, and separate equations for the percentages of monospermic and polyspermic fertilization are obtained. These equations provide an explanation for empirical observations which have shown a decreased percentage of successful egg development at high sperm concentrations. Comparison is made with an earlier heuristic attempt (Styan, 1998) to predict the extent of polyspermic fertilization, and it is found that this earlier method can underestimate the percentage of polyspermic fertilization by up to 10 percent. Moreover, the approach used here retains the flexibility to model changes in sperm concentration due to dispersal mechanisms, and is able to model different mechanisms for the block to polyspermy.


Chronic pain: a long-term sequela of epidermal necrolysis (Stevens-Johnson syndrome/toxic epidermal necrolysis) - prevalence, clinical characteristics and risk factors.
Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are associated with various sequelae. Chronic pain, one of these sequelae, has never been systematically evaluated. To assess the persistence of pain in a single-centre cohort of 113 consecutive patients with SJS/TEN. From this cohort, 81 patients were interviewed more than 1 year after the initial episode and included in the study. Data were collected according to standardized questionnaires. From the 81 interviewed patients, 52 patients (64%) were painless and 29 patients (36%) were painful. Chronic pain syndrome was associated with a more severe initial acute phase of the disease (larger extent of detachment, higher SCORTEN, increased rate of admission in ICU and complications, and longer hospital stay). Pain was mainly located at the level of eyes (55%), mouth and lower limbs (38-41%), with a moderate daily intensity on average (4.7/10). The 'affective' descriptors prevailed over the 'sensory' descriptors, with the exception of burning and itching sensations. Finally, regarding provoked pain, mechanical allodynia (to brushing and pressure) was more marked than thermal allodynia. The persistence of chronic pain after SJS/TEN is a common phenomenon. Sensory descriptors are consistent with sensitization of both small-diameter nerve fibres (burning and itching sensations) and large-diameter nerve fibres (mechanical allodynia), but the affective-emotional components of pain largely predominate. Complex mechanisms lead to persistent pain as long-term sequela of SJS/TEN, among which mechanisms, psychological factors related to post-traumatic stress disorder probably play a key role.


Evolution of phosphorylation-dependent regulation of activation-induced cytidine deaminase.
Interaction of activation-induced cytidine deaminase (AID) with replication protein A (RPA) has been proposed to promote AID access to transcribed double-stranded (ds) DNA during immunoglobulin light chain and heavy chain class switch recombination (CSR). Mouse AID (mAID) interaction with RPA and transcription-dependent dsDNA deamination in vitro requires protein kinase A (PKA) phosphorylation at serine 38 (S38), and normal mAID CSR activity depends on S38. However, zebrafish AID (zAID) catalyzes robust CSR in mouse cells despite lacking an S38-equivalent PKA site. Here, we show that aspartate 44 (D44) in zAID provides similar in vitro and in vivo functionality as mAID S38 phosphorylation. Moreover, introduction of a PKA site into a zAID D44 mutant made it PKA dependent for in vitro activities and restored normal CSR activity. Based on these findings, we generated mAID mutants that similarly function independently of S38 phosphorylation. Comparison of bony fish versus amphibian and mammalian AIDs suggests evolutionary divergence from constitutive to PKA-regulated RPA/AID interaction.


Variation in Twitter posts referencing pediatric sleep aids.
Over 50% of parents who use social media seek parenting advice, however little is known about social media discussions surrounding child sleep aid use. The current study investigated Twitter posts about the use of pediatric sleep aids (melatonin, cannabidiol, weighted blankets, and essential oils), including post frequency, user characteristics, and content. In addition, variation in tweets before and after the onset of the COVID-19 pandemic was examined. Twitter was searched over a 25-month period using TweetDeck. Tweets were coded for user characteristics (eg, affiliations, gender) and content (eg, tone, states sleep or health outcomes, reference to a neurodevelopmental condition). Of the 2754 tweets analyzed, melatonin was referenced most often (60%), followed by essential oils (23%), weighted blankets (14%), and cannabidiol (3%). Most were published by individual users (77%) and were positive (51%) in tone. About 1-third of tweets noted positive sleep or health effects of the sleep aid and only 7% referenced a neurodevelopmental condition. Tweets about pediatric sleep aids increased during the pandemic, primarily those posts about melatonin. Melatonin is the most commonly discussed sleep aid on Twitter, followed by essential oils. Tweets are primarily positive. The number of tweets about sleep aids, specifically melatonin, has increased with time, with significantly more tweets after the start of the pandemic. Clinicians should consider using this outlet to provide empirically-based information regarding the efficacy and benefits or risks of sleep aid use in children.


Mitochondrial spare respiratory capacity: Mechanisms, regulation, and significance in non-transformed and cancer cells.
Mitochondrial metabolism must constantly adapt to stress conditions in order to maintain bioenergetic levels related to cellular functions. This absence of proper adaptation can be seen in a wide array of conditions, including cancer. Metabolic adaptation calls on mitochondrial function and draws on the mitochondrial reserve to meet increasing needs. Among mitochondrial respiratory parameters, the spare respiratory capacity (SRC) represents a particularly robust functional parameter to evaluate mitochondrial reserve. We provide an overview of potential SRC mechanisms and regulation with a focus on its particular significance in cancer cells.


Fulminant Reversible Cerebral Vasoconstriction Syndrome After Carotid Endarterectomy for Asymptomatic Stenosis.
Reversible cerebral vasoconstriction syndrome (RCVS) is a rare complication after carotid endarterectomy (CEA). Only a limited number of prior cases of RCVS have been reported after CEA for asymptomatic carotid stenosis. In this report we present an unusual case of RCVS associated with multifocal intraparenchymal hemorrhage, subarachnoid hemorrhage, subdural hematoma, and ischemic stroke after CEA for asymptomatic carotid stenosis. We review preexisting studies and draw correlations with implications for understanding the mechanisms of RCVS. A high index of suspicion should be maintained in post-CEA patients presenting with headaches or focal neurologic deficits, and vigilance with serial vascular imaging may help minimize long-term complications.


Importance of reasons for stocking adult vaccines.
To identify the most important reasons underlying decisions to stock or not stock adult vaccines. US physicians, nurses, pharmacists, and administrators of internal medicine, family medicine, obstetrics/gynecology, and multispecialty practices who were involved in vaccine stocking decisions (N = 125) completed a best-worst scaling survey online between February and April 2018. Sixteen potential factors influencing stocking decisions were developed based on key informant interviews and focus groups. Respondents selected factors that were most and least important in vaccine stocking decisions. Relative importance scores for the best-worst scaling factors were calculated. Survey respondents described which vaccines their practice stocks and reasons for not stocking specific vaccines. Subgroup analyses were performed based on the respondent's involvement in vaccine decision making, role in the organization, specialty, and affiliation status, as well as practice characteristics such as practice size, insurance mix, and patient age mix. Relative importance scores for stocking vaccines were highest for "cost of purchasing vaccine stock," "expense of maintaining vaccine inventory," and "lack of adequate reimbursement for vaccine acquisition and administration." Most respondents (97%) stocked influenza vaccines, but stocking rates of other vaccines varied from 39% (meningococcal B) to 83% (tetanus-diphtheria-pertussis). Best-worst scaling results were consistent across respondent subgroups, although the range of vaccine types stocked differed by practice type. Economic factors associated with the purchase and maintenance of vaccine inventory and inadequate reimbursement for vaccination services were the most important to decision makers when considering whether to stock or not stock vaccines for adults.


Association between hypertriglyceridemic-waist phenotype and non-alcoholic fatty liver disease: a general population-based study.
Hypertriglyceridemic-waist (HTGW) phenotype has been proposed as a practical tool for screening the risk of cardiovascular diseases and glycemic metabolic disease. This study sought to investigate the relationship between HTGW phenotype and non-alcoholic fatty liver disease (NAFLD). A total of 14,251 subjects who took part in health screening were enrolled in the study and NAFLD was diagnosed by abdominal ultrasound. According to triglyceride (TG) and waist circumference, the study population was divided into four phenotypes, in which HTGW phenotype was defined as TG ≥ 1.7 mmol/L and male waist circumference ≥ 90 cm or female waist circumference ≥ 80 cm. Multivariate logistic regression analysis was used to evaluate the relationship between HTGW phenotype and NAFLD. In the current study, 2.43% of the subjects had HTGW phenotype, while the prevalence of NAFLD in subjects with HTGW phenotype was 77.81%. After full adjustment for covariates, compared with people with normal waist circumference and TG levels, the risk of NAFLD in people with normal TG levels but enlarged waist circumference increased by 39% [OR:1.39, 95%CI: 1.15, 1.68], in people with normal waist circumference but elevated TG levels increased by 96% [OR:1.96, 95%CI: 1.65, 2.33], and in subjects with HTGW phenotype increased by 160% [OR:2.60, 95%CI: 1.88, 3.58]. Additionally, further analysis suggested that there were significant interactions between age, height, BMI and NAFLD risk associated with TGW phenotypes. Receiver operating characteristic curves analysis suggested that the combination of TG and waist circumference further improved the diagnostic value for NAFLD. HTGW phenotype is associated with NAFLD risk in the general population, which may be a novel and accessible indicator for NAFLD screening.


Deletion of the progranulin gene in patients with frontotemporal lobar degeneration or Parkinson disease.
Progranulin gene (PGRN) mutations cause ubiquitin-positive frontotemporal lobar degeneration linked to chromosome 17 (FTLDU-17). The spectrum of known mutations strongly suggests that neurodegeneration results from a partial loss of PGRN function and leads to the hypothesis that PGRN gene deletions could be present in a subset of patients. We analysed 63 unrelated French patients with frontotemporal lobar degeneration (FTLD) for PGRN gene dosage alteration by quantitative multiplex PCR of short fluorescent fragments (QMPSF). We identified in one patient with typical PGRN neuropathology a near complete deletion, removing exons 1-11, of the PGRN gene. This deletion, which resulted from a nonhomologous recombination event, was equally present in one affected sister presenting with Parkinson disease (PD). This observation provides a final argument that PGRN mutations exert their pathogenic effect through haploinsufficiency and underlines the diversity of clinical presentations associated with these PGRN alterations.


Genetics of dental agenesis: anterior and posterior area of the arch.
The aim of this study was to evaluate whether or not dental agenesis is regulated by genes and, in those cases under genetic control, if there were differences between incisors-canines and premolars-molars in the genetic transmission of agenesis. This study was conducted on all patients who presented for a clinical evaluation in the Orthodontic Unit of the Department of Dentistry between 2001 and 2002. All patients were screened for dental agenesis. At the end of the selection process, 17 patients with dental agenesis, 6 to 20 years of age, were questioned about their family dental history for the purpose of constructing family trees and 15 family trees were constructed. The patients were questioned regarding any dental anomalies present. Panoramic radiographs, casts, and dental examinations were performed on the patients. For the patient's relatives, panoramic radiographs and oral histories were generally collected. The most often missing teeth were mandibular second premolars. On the basis of the family trees, the data allowed for the identification of two different groups: 1) 9 families had dental agenesis as a function of genetic transmission and 2) 6 families exhibited dental agenesis as a sporadic condition. In the former group of patients, agenesis primarily involved the maxillary lateral incisors. This appeared to be transmitted as an autosomal dominant characteristic, with variable expression and incomplete penetrance. In these families, both in individuals with and without agenesis, other anomalies were present, such as supernumerary teeth, microdontia, and anomalous teeth. In these cases, agenesis was one of the manifestations of an anomaly of the dental lamina, specifically mild dysplasia. In the latter group of patients, agenesis was a characteristic present only in orthodontic patients. In this group, agenesis most commonly involved the second molars and the second premolars. These findings indicate that agenesis of anterior teeth may depend on genes, while agenesis of posterior teeth is sporadic.


Amino acid sequences of stomach and nonstomach lysozymes of ruminants.
Complete amino acid sequences are presented for lysozymes c from camel and goat stomachs and compared to sequences of other lysozymes c. Tree analysis suggests that the rate of amino acid replacement went up as soon as lysozyme was recruited for the stomach function in early ruminants. The two lysozymes from goat stomach are the products of a gene duplication that probably took place before the divergence of cow, goat, and deer about 25 million years ago. Partial sequences of three lysozymes from goat tears indicated that (a) the goat tear family of lysozymes may have diverged from the stomach lysozyme family by an ancient duplication and (b) later duplications are probably responsible for the multiple forms of tear and milk lysozymes in ruminants.


Ewing's family of tumors in adults: multivariate analysis of survival and long-term results of multimodality therapy in 182 patients.
To assess the outcome and the prognosis of adults with a neoplasm related to the Ewing's sarcoma family of tumors. The outcomes of 182 consecutive patients older than 15 years with Ewing's sarcoma or related neoplasms managed from 1982 to 1992 were reviewed, without any selection according to primary tumor site or disease extension. Of 182 patients, 53 had evidence of metastases at presentation (29%). Tumor size was greater than 10 cm in 70 patients (41%). With a median follow-up duration of 66 months, the 5-year overall survival (OS) rate was 41%. In patients with localized disease, 5-year OS rate was 54% and 5-year progression-free survival (PFS) rate, 43%. Late relapses after 5 years accounted for 9% of relapses. Metastasis at presentation (P = .00001), pelvic primary lesion (P = .0025), and tumor size greater than 10 cm (P = .004) were independent prognostic factors for survival. Five-year OS was 67% in patients with nonpelvic tumors < or = 10 cm, 52% in those with pelvic tumors less than 10 cm or extrapelvic tumors > or = 10 cm, 16% in those with pelvic tumors greater than 10 cm, and 9% in those with metastasis (P = .00001). Based on our experience and a review of the literature, we concluded that the natural history and the prognosis of the Ewing's family of tumors in adults are not different from that found in children. A greater tumor bulk in adults may explain the less favorable prognosis previously reported by others. Outcome could be adequately monitored by a simple prognostic index.


Left-handed mirror writing following right anterior cerebral artery infarction: evidence for nonmirror transformation of motor programs by right supplementary motor area.
A patient with acquired, pathologic left-handed mirror writing and mirror movements during bimanual coordination is reported. CT demonstrated an ischemic infarction involving the right supplementary motor area, medial prefrontal cortex, and anterior cingulate gyrus with nearly total sparing of the corpus callosum. This case, together with recent experimental findings in monkeys, supports the hypothesis that the supplementary motor area may be responsible for nonmirror transformation of motor programs originating in the left hemisphere prior to execution by the primary motor area in the right hemisphere.


In vivo monitoring of acetylcholine release from cardiac vagal nerve endings in anesthetized mice.
We applied a microdialysis technique to the left ventricular myocardium of anesthetized mice and tried to monitor acetylcholine (ACh) release from cardiac vagal nerves. Transection of bilateral cervical vagal nerves decreased dialysate ACh concentration. Electrical stimulation of the left cervical vagal nerve increased dialysate ACh concentration in proportion to the frequency of stimulation. Intravenous administration of hexamethonium, prevented the increase in dialysate ACh concentration during vagal nerve stimulation, indicating that ACh in the dialysate primarily reflects ACh released from post-ganglionic cardiac vagal nerves. Microdialysis permits monitoring of ACh release from post-ganglionic cardiac vagal nerves that are most likely to be innervating the left ventricle in mice.


Disturbing behaviors in demented elders: a concept synthesis.
This article describes the use of concept synthesis, as defined by Walker and Avant, to extract meaningful syndromes of disturbing behaviors as they are manifested in demented elders. The process resulted in the identification of five distinct behavioral clusterings: Aggressive Psychomotor Behavior, Nonaggressive Psychomotor Behavior, Verbally Aggressive Behavior, Passive Behavior and Functionally Impaired Behavior. These synthesized concepts are an initial attempt at clarifying nursing perspective on disturbing behaviors in dementia.


Radioimmunoassay of serum prostatic acid phosphatase after prostatic massage.
The effect of prostatic massage on the concentration of prostatic acid phosphatase (PAP) in blood serum as determined by radioimmunoassay (RIA) was compared with that determined by a standard enzymatic assay (EA). Serum was drawn from 24 men before prostatic massage and after--at specified intervals, up to twenty-four hours. Three of these men were young, normal controls; 10 had biopsy-proved prostate cancer (CA); 11 had histologically confirmed benign prostatic hyperplasia (BPH). After prostatic massage, 3 of the 10 CA patients (30%) had elevation of PAP as determined by EA and 4 of the 11 BPH patients (36%) as determined by RIA. None of the controls showed elevated levels of PAP by either assay. In all patients elevated levels of PAP by both assays had returned to normal twenty-four hours after massage. It was concluded that serum for PAP testing by either assay method should be drawn before or twenty-four hours after rectal examination to prevent false positive results and the need for retesting.


Low polarity ligands of sex hormone-binding globulin in pregnancy. Part II--Identification.
Certain previously unrecognized ligands of SHBG of low polarity in pregnancy were identified. They include two weakly bound compounds: 5 alpha-pregnane-3,20-dione and progesterone; and two strongly bound substances, 2-methoxyestrone and a new steroid, estradienolone (17 beta-hydroxy-1,5-estradiene-3-one). The identification of the first three peaks was based on chromatographic elution patterns, binding characteristics and gas chromatography-mass spectrometry. The identification of the fourth peak, the new steroid, was based on similar kinds of evidence and, in addition, solubility characteristics and ultraviolet absorption spectrum.


Phospholipases, enzymes that share a substrate class.
Considerable work has gone into the study of PLs since the first suggestions of their existence nearly a century ago. This work has intensified enormously since the mid-1970s when their role in signal-coupling mechanisms and in pathophysiology was recognized. While much has been done to understand this diverse group of enzymes at the molecular and mechanistic levels, the discovery of new PLs has far outstripped our capacity to study them in sufficient detail to appreciate what makes each unique while perhaps having some common mechanisms of action and regulation. One would almost plead: No new PLs - Let us study those at hand! That is not the case in our field and the discovery of new PLs will continue. It is important, however, that an understanding be gained of these enzymes at the molecular level, how they interact with their substrates, and how regulatory factors can target the function of PLs in situ.


MIR31HG exhibits oncogenic property and acts as a sponge for miR-361-3p in cervical carcinoma.
The long noncoding RNA (lncRNA) MIR31 Host Gene (MIR31HG) is a crucial regulator in malignant cancers. In this work, I examined the potential function and molecular mechanism of MIR31HG in cervical carcinoma progression. MIR31HG was strikingly upregulated in clinical cervical carcinoma specimens compared with that in adjacent samples. Functionally, knockdown of MIR31HG noticeably repressed cervical carcinoma cell growth and invasiveness in vitro and inhibited cervical carcinoma cell growth in vivo. Mechanistically, MIR31HG was identified as an endogenous 'sponge' through competing for miR-361-3p binding to modulate the miRNA target, epithelial membrane protein 1 (EMP1). Lastly, I verified that overexpression of EMP1 or miR-361-3p silencing effectively offset the impacts of MIR31HG knockdown on cervical carcinoma cell progression. In brief, these experimental results validate that MIR31HG is an oncogenic lncRNA that facilities cervical cancer progression.


Clinical and epidemiological characteristics associated with unfavorable tuberculosis treatment outcomes in TB-HIV co-infected patients in Brazil: a hierarchical polytomous analysis.
TB patients co-infected with HIV have worse treatment outcomes than non-coinfected patients. How clinical characteristics of TB and socioeconomic characteristics influence these outcomes is poorly understood. Here, we use polytomous regression analysis to identify clinical and epidemiological characteristics associated with unfavorable treatment outcomes among TB-HIV co-infected patients in Brazil. TB-HIV cases reported in the Brazilian information system (SINAN) between January 1, 2001 and December 31, 2011 were identified and categorized by TB treatment outcome (cure, default, death, and development of MDR TB). We modeled treatment outcome as a function of clinical characteristics of TB and patient socioeconomic characteristics by polytomous regression analysis. For each treatment outcome, we used cure as the reference outcome. Between 2001 and 2011, 990,017 cases of TB were reported in SINAN, of which 93,147 (9.4%) were HIV co-infected. Patients aged 15-19 (OR=2.86; 95% CI: 2.09-3.91) and 20-39 years old (OR=2.30; 95% CI: 1.81-2.92) were more likely to default on TB treatment than those aged 0-14 years old. In contrast, patients aged ≥60 years were more likely to die from TB (OR=2.22; 95% CI: 1.43-3.44) or other causes (OR=2.86; 95% CI: 2.14-3.83). Black patients were more likely to default on TB treatment (OR=1.33; 95% CI: 1.22-1.44) and die from TB (OR=1.50; 95% CI: 1.29-1.74). Finally, alcoholism was associated with all unfavorable outcomes: default (OR=1.94; 95% CI: 1.73-2.17), death due to TB (OR=1.46; 95% CI: 1.25-1.71), death due to other causes (OR=1.38; 95% CI: 1.21-1.57) and MDR-TB (OR=2.29; 95% CI: 1.46-3.58). Socio-economic vulnerability has a significant effect on treatment outcomes among TB-HIV co-infected patients in Brazil. Enhancing social support, incorporation of alcohol abuse screening and counseling into current TB surveillance programs and targeting interventions to specific age groups are interventions that could improve treatment outcomes.


Prevalence, infectivity and correlates of hepatitis B virus infection among pregnant women in a rural district of the Far North Region of Cameroon.
Epidemiological data on hepatitis B virus (HBV) infection among pregnant women in Cameroon are very scarce, especially in the rural milieu. The purpose of this study was to determine the prevalence and factors associated with HBV infection, and the infectivity of rural pregnant women in the Far North Region of Cameroon. A cross-sectional study was conducted in three rural health facilities of the Guidiguis health district between December 2013 and March 2014. We consecutively recruited 325 pregnant women attending antenatal consultations. A pretested questionnaire was used to collect socio-demographic data and factors associated with HBV infection. The presence of hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and human immunodeficiency virus (HIV) were determined using commercial test strips. Regression analyses were used to assess correlates of HBV infection. The mean age was 24.4 (SD5.6) years. Most women were married (97.2%) and housewives (96.4%), with less than secondary education level (80%). Only 4 women (1.2%) had been vaccinated against HBV. Thirty-three women (10.2%) were HBsAg-positive, of whom 4 (12.1%) were positive to HBeAg. The prevalence of HIV infection was 2.5% (8/325). Overall, 5 (1.5%) women were co-infected with HIV and HBV. Independent correlates of HBV infection included history of blood transfusion (adjusted odd ratio 12.59, 95% CI 1.46-108.89; p = 0.021) and concurrent infection by HIV (adjusted odd ratio 22.53, 95% CI 4.76-106.71; p < 0.0001). The prevalence of HBV infection among pregnant women in this rural milieu is high. History of blood transfusion and HIV infection are highly associated with HBV infection. The relative low rate of women positive to both HBsAg and HBeAg suggests that perinatal transmission of HBV might not be the prevailing mode of HBV transmission in this area.


Higher glycolytic capacities in muscle of carnivorous rainbow trout juveniles after high dietary carbohydrate stimulus at first feeding.
Rainbow trout is a "glucose-intolerant" carnivorous species. Using the metabolic programming strategy, we used early nutritional stimuli in order to modify carbohydrate utilization in trout juveniles. Fish were fed two diets during the first feeding, namely HP (no carbohydrate / high protein) diet and LP (high carbohydrate / low protein) diet. HP diet was used as the control diet and LP diet as an early stimulus diet. We also used another early stimulus with fish fed HP diet every other day during the first feeding (HP restriction feeding - HPR). After the first-feeding stage (4 weeks), all fish were subsequently subjected to a growth trial with a commercial diet followed by a challenge test with the LP diet (11 weeks). At the end of the first feeding stimulus and of the challenge test, we investigated growth performance, glucose metabolism-related parameters and global DNA CmCGG methylation in trout. LP and HPR dietary stimuli have been a success as shown by the direct modifications of growth performance and mRNA levels for glucose metabolism-related genes at the end of first feeding compared to alevins fed the HP diet. At the end of the challenge trial, no variation in growth performance and hepatic metabolism of LP-history and HPR-history in trout juveniles were observed. However, in muscle of trout juvenile subjected to LP diet at the first feeding, we found an up-regulation of mRNA levels of some glucose metabolism (glucose transport and glycolysis)-related genes and an increase of activities of important glycolysis-related enzymes (hexokinase, phosphofructokinase and pyruvate kinase). These observations are associated with a decrease in the content of glycogen compared to fish fed the HP diet. Moreover, global CmCGG DNA methylation in the muscle of fish with LP history was significantly lower than those fed the HP diet. Dietary LP stimulus at first feeding could permanently modify glucose metabolism and global CmCGG DNA methylation level in muscle of trout juveniles, showing that the first feeding stage is efficient for programming the glucose metabolism in fish.


Construction and evaluation of two computational models for predicting the incidence of influenza in Nagasaki Prefecture, Japan.
It remains challenging to forecast local, seasonal outbreaks of influenza. The goal of this study was to construct a computational model for predicting influenza incidence. We built two computational models including an Autoregressive Distributed Lag (ARDL) model and a hybrid model integrating ARDL with a Generalized Regression Neural Network (GRNN), to assess meteorological factors associated with temporal trends in influenza incidence. The modelling and forecasting performance of these two models were compared using observations collected between 2006 and 2015 in Nagasaki Prefecture, Japan. In both the training and forecasting stages, the hybrid model showed lower error rates, including a lower residual mean square error (RMSE) and mean absolute error (MAE) than the ARDL model. The lag of log-incidence, weekly average barometric pressure, and weekly average of air temperature were 4, 1, and 3, respectively in the ARDL model. The ARDL-GRNN hybrid model can serve as a tool to better understand the characteristics of influenza epidemic, and facilitate their prevention and control.


Prevention of irradiation-induced salivary hypofunction by microvessel protection in mouse salivary glands.
Treatment of most head and neck cancers includes radiotherapy. Salivary glands (SGs) in the irradiation (IR) field are irreversibly damaged resulting in severe hyposalivation. We evaluated the importance of SG endothelial cells to this IR-induced injury, and whether serotype 5 adenoviral (Ad5) vector-mediated transfer of basic fibroblast growth factor (AdbFGF) or vascular endothelial growth factor (AdVEGF) complementary DNAs would afford radioprotection. Four hours after IR, microvessel density (MVD) in SGs decreased by approximately 45%. However, if mice were pre-treated with either AdVEGF or AdbFGF 48 hours before IR the loss in MVD was significantly reduced. An irrelevant vector, AdLacZ, encoding Escherichia coli beta-galactosidase, was without effect. After 8 weeks, IR reduced salivary flow by approximately 65% in untreated mice. Mice pre-treated (using 5 x 10(9) particles/gland 48 hours prior to IR) with AdLacZ exhibited a reduction in salivary flow similar to untreated mice receiving IR. However, irradiated mice pre-treated with AdbFGF or AdVEGF showed a significant improvement in their salivary flow, to approximately 70% (P < 0.01) and 80% (P < 0.01), respectively, compared to non-irradiated control mice. These results are consistent with the notion that injury to the adjacent microvasculature may play an important role in SG radiation damage. Furthermore, our results suggest that a local transient treatment directed at protecting SG endothelial cells may be beneficial for patients undergoing IR for head and neck cancer.


Oral bioaccessibility of silver nanoparticles and ions in natural soils: Importance of soil properties.
The abundance of silver nanoparticles (AgNPs) in consumer products has led to their environmental release and therefore to concern about their impact on human health. The ingestion of AgNP-contaminated soil from urban sites is an important exposure pathway, especially for children. Given the limited information on oral bioaccessibility of soil Ag, we used a physiologically based extraction test (PBET) to evaluate the bioaccessibility of AgNPs and AgNO3 from soil digestion. The AgNPs underwent several biochemical transformations, including their simultaneous dissolution and agglomeration in gastric fluid followed by the disintegration in the intestinal fluid of the agglomerates into NPs containing silver and chlorine. Therefore, Ag-containing soil exposed the intestine to nanoparticulate Ag in forms that were structurally different from the original forms. The bioaccessibility of AgNPs (0.5 ± 0.05%-10.9 ± 0.7%) was significantly lower than that of AgNO3 (4.7 ± 0.6%-14.4 ± 0.1%), as a result of the lower adsorption of nanoparticles to soil residues during the digestive process. For the soils tested, the bioaccessibility of AgNPs increased with decreasing clay contents and lower pH. By identifying the soil properties that control AgNP bioaccessibility, a more efficient and accurate screening can be performed of soil types that pose the greatest health risk associated with AgNP exposure.


Effect of antioxidants on sperm genetic damage.
According to worldwide statistics, between one in four and one in five couples have fertility problems. These problems are equally distributed between males and females. Modern lifestyle has obviously increased these problems: endocrine-disrupting chemicals, such as plastic polymer catalysts, alkylphenols, phthalates and so on, and cosmetic additives seem to be strongly involved in this fertility problem. Many of these compounds increase oxidative stress (OS) and thus impair spermatogenesis. The oocyte has only a finite capacity, decreasing with maternal age, to repair sperm-borne decays. To decrease this DNA repair burden, reducing the sperm DNA damages linked to OS is tempting. Antioxidant vitamins are often given haphazardly; they are not very efficient and potentially detrimental. A detailed analysis of the sperm nucleus is mandatory (DNA fragmentation or lack of nuclear condensation) prior to any treatment. Here we discuss new concepts in OS and the corresponding therapeutic approaches.


Suppression of AMP-activated protein kinase reverses osteoprotegerin-induced inhibition of osteoclast differentiation by reducing autophagy.
Osteoclasts (OC) are unique terminally differentiated cells whose primary function is bone resorption. We previously showed that osteoprotegerin (OPG) inhibits OC differentiation in vitro by enhancing autophagy via the adenosine monophosphate-activated protein kinase (AMPK)/mTOR/p70S6K signalling pathway in vitro. Here, we aimed to elucidate the mechanism of AMPK mediated autophagy to regulate OPG-mediated inhibition of OC differentiation and identify potential therapeutic targets associated with bone loss. We used the AMPK activator AICAR to determine the relationship between AMPK activation and OC differentiation, and studied the role of AMPK-mediated autophagy in OPG-mediated inhibition of OC differentiation by using autophagy inhibitors or AMPK knockdown. AMP-activated protein kinase activation caused LC3II accumulation and weakened OC differentiation activity. In contrast, inactivation of autophagy by 3-methyladenine or Bafilomycin A1 could attenuate OPG-mediated inhibition of OC differentiation via the AMPK/mTOR/p70S6K signalling pathway. Furthermore, the AMPK inhibitor compound C and knockdown of AMPK impaired OPG-mediated inhibition of OC differentiation by inducing autophagy. These results demonstrated that the AMPK signalling pathway functions as a critical regulator in the OPG-mediated inhibition of OC differentiation, by inducing autophagy. Our results provide a basis for future bone-related studies on the AMPK signalling pathway.


Impact of fertilization on chestnut growth, N and P concentrations in runoff water on degraded slope land in South China.
Growing fruit trees on the slopes of rolling hills in South China was causing serious environmental problems because of heavy application of chemical fertilizers and soil erosion. Suitable sources of fertilizers and proper rates of applications were of key importance to both crop yields and environmental protection. In this article, the impact of four fertilizers, i.e., inorganic compound fertilizer, organic compound fertilizer, pig manure compost, and peanut cake (peanut oil pressing residue), on chestnut (Castanea mollissima Blume) growth on a slope in South China, and on the total N and total P concentrations in runoff waters have been investigated during two years of study, with an orthogonal experimental design. Results show that the organic compound fertilizer and peanut cake promote the heights of young chestnut trees compared to the control. In addition, peanut cake increases single-fruit weights and organic compound fertilizer raises single-seed weights. All the fertilizers increased the concentrations of total N and total P in runoff waters, except for organic compound fertilizer, in the first year experiment. The observed mean concentrations of total N varied from 1.6 mg/L to 3.2 mg/L and P from 0.12 mg/L to 0.22 mg/L, which were increased with the amount of fertilizer applications, with no pattern of direct proportion. On the basis of these experiment results, organic compound fertilizer at 2 kg/tree and peanut cake at 1 kg/tree are recommended to maximize chestnut growth and minimize water pollution.


Torpedo synaptophysin: evolution of a synaptic vesicle protein.
Synaptophysin is an integral membrane protein of synaptic vesicles found in neurons and endocrine cells. Synaptophysin monomers associate into hexamers forming a large conductance channel. We present an analysis of synaptophysin from the nervous system of the marine ray Torpedo californica. Analysis of cDNA clones reveals a 62% amino acid similarity between the Torpedo and rat sequences. The 4 hydrophobic membrane spanning domains and the glycosylation site are conserved. In contrast, the two intravesicular loops connecting the membrane spanning regions, show varying degrees of sequence conservation, suggesting that portions of these domains may play critical functional roles. The carboxyterminal tail has been proposed to bind calcium and is a major site for tyrosine phosphorylation. The precise sequence of this region has almost completely diverged while the proline-tyrosine rich nature is maintained. Blotting studies reveal the RNA and the protein in nervous system tissues and demonstrate that the molecule copurifies with cholinergic synaptic vesicles.


Laparoscopic pouch resizing and redo of gastro-jejunal anastomosis for pouch dilatation following gastric bypass.
With a dramatically increasing number of bariatric operations performed world-wide in the recent years, more late complications have been noticed. Proximal gastric pouch dilatation is a known late complication after laparoscopic or open restrictive surgery for morbid obesity. In the present paper, we report our experience with laparoscopic re-operation of enlarged gastric pouches after laparoscopic gastric bypass, with emphasis on technique and outcome. Data were retrieved from a prospective database of 334 patients who underwent a laparoscopic gastric bypass operation at the University Hospital of Zurich from July 2000 to December 2004. Five laparoscopic revisions for pouch dilatation after primary bypass were performed. 3 female and 2 male patients with median age 40 years (range 32-55) underwent a laparoscopic pouch resizing. At the time of the re-operation, the median BMI was 32.0 kg/m(2) (range 28.4-48.4). All procedures were performed laparoscopically with no conversion to open surgery. The median operating-time was 110 minutes (95-120). The median hospital stay was 6 days (range 5-14). The median BMI in the follow-up of 12 months (9-14) was 28.0 kg/m(2) (25.5-45.8). Diabetes mellitus improved in 4 cases during follow-up. Laparoscopic pouch resizing with redo of the gastro-jejunal anastomosis was feasible, safe and effective in this small series. It led to further weight loss and improved symptoms of poor pouch emptying.


Implementing a probabilistic definition of freedom from infection to facilitate trade of livestock: putting theory into praxis for the example of bovine herpes virus-1.
International trade of livestock and livestock products poses a significant potential threat for spread of diseases, and importing countries therefore often require that imported animals and products are free from certain pathogens. However, absolute freedom from infection cannot be documented, since all test protocols are imperfect and can lead to false-negative results. It is possible instead to estimate the "probability of freedom from infection" and its opposite, the probability of infection despite having a negative test result. These probabilities can be estimated based on a pre-defined target prevalence, known surveillance efforts in the target population and known test characteristics of any pre-export test. Here, calculations are demonstrated using the example of bovine herpes virus-1 (BoHV-1). In a population that recently became free of BoHV-1 without using vaccination, the probability of being infected of an animal randomly selected for trade is 800 per 1 million and this probability is reduced to 64 (95% probability interval PI 6-161) per 1 million when this animal is tested negatively prior to export with a gB-ELISA. In a population that recently became free of BoHV-1 using vaccination, the probability of being infected of an animal randomly selected for trade is 200 per 1 million, and this probability can be reduced to 63 (95% PI 42-87) when this animal is tested negatively prior to export with a gE-ELISA. Similar estimations can be made on a herd level when assumptions are made about the herd size and the intensity of the surveillance efforts. Subsequently, the overall probability for an importing country of importing at least 1 infected animal can be assessed by taking into account the trade volume. Definition of the acceptable level of risk, including the probability of false-negative results to occur, is part of risk management. Internationally harmonized target prevalence levels for the declaration of freedom from infection from selected pathogens provide a significant contribution to the facilitation of international trade of livestock and livestock products by allowing exporting countries to design tailor-made output-based surveillance programs, while providing equivalent guarantees regarding the probability of freedom from infection of the population. Combining this with an approach to assess the overall probability of introducing at least 1 infected animal into an importing country during a defined time interval will help importing countries to achieve their desired level of acceptable risk and will help to assess the equivalence of animal health and food safety standards between trading partners.


Mixed outbreak of parainfluenza type 1 and influenza B associated with tourism and air travel.
A preseasonal outbreak of influenza-like illness was reported in a tourist group that had returned from Ireland to the United States on October 1 and 2, 1999. The authors investigated to determine the timing, extent, and nature of the outbreak. A cohort study was conducted among the entire group. Cases were defined as illness occurring among tour members who experienced cough or sore throat with onset from September 29 through October 5. Nasopharyngeal and throat cultures were examined for respiratory viruses. Eighteen (60%) of the 30 tourists were ill; the majority (66%) of cases had onset of illness within 1 day of departure from Ireland. Human parainfluenza virus type 1 was isolated from six cultures, and influenza B from three. The attack rate was 100% among the eight persons 65 years of age or older versus 45% among the 22 persons younger than 65 years (P = 0.01). International travellers, particularly older persons and members of organized tour groups, may experience increased risks for respiratory viral infection. The recognition and containment of imported infectious diseases depend on prompt reporting and epidemiologic investigation.


Clinical significance of neutrophil gelatinase-associated lipocalin and sdLDL-C for coronary artery disease in patients with type 2 diabetes mellitus aged ≥ 65 years.
Although type 2 diabetes mellitus (T2DM) and coronary artery disease (CAD) share many common pathological and physiological characteristics, there are few studies assessing the predictive capacity of novel biomarkers in occurrence and development of CAD in T2DM patients aged ≥ 65 years. In addition, T2DM patients aged ≥ 65 years are prone to CAD. Therefore, it is of great significance to find novel biomarkers for the development CAD in T2DM. In this retrospective cohort study, 579 T2DM patients aged ≥ 65 years were consecutively enrolled in this work, and 177 of whom had major adverse cardiovascular and cerebrovascular events (MACCE: cardiovascular or cerebrovascular death, acute coronary syndrome, coronary stent implantation, and stroke) during the follow up. Univariate and multivariate factors were employed to analyze the correlation between each variable and the occurrence of MACCE, and the Spearman's rank correlation analysis was performed to assess the relationships between Neutrophil gelatinase-associated lipocalin (NGAL) and small dense low-density lipoprotein-cholesterol (LDL-C) (sdLDL-C). The receiver operating characteristic (ROC) curve was adopted to determine the predictive value of NGAL and sdLDL-C elevation for MACCE in T2DM patients aged ≥ 65 years. After a median 48 months follow-up [19, (10 ~ 32) ], the levels of NGAL, sdLDL-C, hemoglobin A1c (HbA1c), LDL-C, and apolipoprotein B (ApoB) were significantly higher while those of high-density lipoprotein cholesterol (HDL-C) and apolipoprotein A I (ApoA-I) were lower in MACCE positive group. NGAL correlated to body mass index (BMI) (r = 0.391, P = 0.001) and triglyceride (TG) (r = 0.228, P = 0.032), and high-sensitivity CRP (hsCRP) (r = 0.251, P = 0.007), and neutrophils (r = 0.454, P = 0.001), sdlDL-C level was found to be positively correlated with LDL-C (r = 0.413, P = 0.001), TG (r = 0.432, P = 0.001), and ApoB (r = 0.232, P = 0.002); and it was negatively correlated with HDL-C (r = -0.362, P = 0.031) and ApoA-I (r = -0.402, P = 0.001). Age-adjusted Cox regression analysis showed that NGAL (HR = 1.006, 95% confidence interval (CI): 1.005-1.008, P < 0.001) and sdLDL-C (HR = 1.052, 95% CI: 1.037-1.066, P < 0.001) were independently associated with occurrence of MACCE. ROC curve analysis showed that NGAL (area under ROC (AUC) = 0.79, 95% CI: 0.75-0.84, P < 0.001) and sdlDL-C (AUC = 0.76, 95% CI: 0.72-0.80, P < 0.001) could predict the occurrence of MACCE (area under ROC. NGAL combined with sdlDL-C could predict the occurrence of MACCE well (AUC = 0.87, 95% CI: 0.84-0.90, P < 0.001). The higher NGAL and sdLDL-C in T2DM patients aged ≥ 65 years were significantly and independently associated with the risk of MACCE, and showed higher clinical values than other lipid biomarkers or other chronic inflammation, so they were expected to be the most effective predictors of MACCE assessment.


Genetic transformation of the symbiotic basidiomycete fungus Hebeloma cylindrosporum.
The pAN7.1 plasmid containing the E. coli hygromycin B phosphotransferase gene was used to transform protoplasts of the ectomycorrhizal fungus Hebeloma cylindrosporum. Hygromycin-resistant transformants were selected at a frequency of one to five per micrograms of transforming DNA. Southern blot analyses revealed multiple copy integration of the transforming plasmid into the genome. The selection system was used to introduce other genes of interest by co-transformation. Two plasmids, one containing tryptophan biosynthesis genes and the other the NADP-glutamate dehydrogenase gene from the saprophytic basidiomycete Coprinus cinereus, were successfully introduced into the H. cylindrosporum genome with up to 70% efficiency of co-transformation. The hygromycin resistance phenotype was stably maintained during growth of transformants on hygromycin-free medium. All transformants retained their ability to form mycorrhizae with the habitual host plant Pinus pinaster, making them suitable for future physiological studies.


Major Shifts in Outpatient Cirrhosis Care Delivery Attributable to the COVID-19 Pandemic: A National Cohort Study.
The coronavirus disease 2019 (COVID-19) pandemic has disrupted health care delivery in the United States, with increased reliance on telemedicine visits as opposed to in-person outpatient appointments. We used national data to evaluate shifts in modes of hepatology outpatient care for patients with cirrhosis during the pandemic. This was a retrospective cohort study among U.S. veterans with cirrhosis. We used linear regression to evaluate absolute and percentage changes from baseline in hepatology in-person visits and telemedicine visits from January 1, 2020, to August 11, 2020. The proportion of in-person and telemedicine visits were plotted geographically to demonstrate state-level shifts in care delivery over time. Patient-level characteristics in the pre-COVID and during-COVID periods were also compared. We identified 5,618 in-person and 6,210 telemedicine hepatology visits among patients with cirrhosis. In-person visits significantly declined (-16.0% per week; 95% confidence interval [CI] -20.7, -11.2; P < 0.001), while telemedicine visits significantly increased (61.3% per week; 95% CI 45.1, 77.5; P < 0.001) in the early during-COVID period. At the U.S. state level, we found that nearly all states experienced a significant shift toward telemedicine over the course of several weeks. Patients over the age of 70 years and Black patients were less likely to receive telemedicine visits in the pre-COVID period (each P < 0.05), although these differences were eliminated in the during-COVID periods. Conclusion: Among patients with cirrhosis, hepatology outpatient care delivery has shifted heavily toward telemedicine due to COVID-19. This occurred across the United States, and changes have been sustained through August 2020. Expanded telemedicine visits among older patients and Black patients may reflect dedicated efforts to increased access to care among these groups.


Characteristics and risk factors associated with missed diagnosis in patients with smear-negative pulmonary tuberculosis.
Delayed diagnosis and treatment of smear-negative pulmonary tuberculosis (TB) are major concerns for TB control. We evaluated characteristics of patients with smear-negative pulmonary TB who received a delayed diagnosis and identified risk factors that may have contributed to this delay. We reviewed medical records of patients with smear-negative culture-positive pulmonary TB treated at a tertiary care hospital in South Korea between January 2017 and December 2018. Patients who initiated anti-TB treatment after positive cultures were included in the missed TB group, and those who initiated empirical treatment before positive cultures were included in the control group. Of 220 patients included, 117 (53.2%) and 103 (46.8%) were in the missed TB and control groups, respectively. Patients in the missed TB group were older (p = 0.001) and had a higher mean body mass index (BMI) (p = 0.019). Comorbidities (66.9% vs. 46.6%, p = 0.003) and immunocompromised patients (33.1% vs. 20.4%, p = 0.035) were more common in the missed TB group than in the control group. Old age (odds ratio [OR], 1.030; 95% confidence interval [CI], 1.012 to 1.048; p = 0.001), high BMI (OR, 1.114; 95% CI, 1.004 to 1.237; p = 0.042), and negative polymerase chain reaction (PCR) results (OR, 9.551; 95% CI, 4.925 to 18.521; p < 0.001) were associated with delayed diagnosis. In more than half of patients with smear-negative pulmonary TB, the diagnosis was delayed. Patients with delayed TB diagnosis were older, had higher BMI, and negative PCR results.


Falls among elderly persons in Latin America and the Caribbean and among elderly Mexican-Americans.
To estimate the prevalence of and risk factors for falls among community-dwelling elders in Latin America and the Caribbean and among elderly Mexican-Americans in the southwestern United States. Data for the study came from a project called Health, Well-Being, and Aging in Latin America and the Caribbean (Salud, Bienestar y Envejecimiento en América Latina y el Caribe) (the "SABE project") (surveys from seven cities, with a total of 9,765 subjects) and from the Hispanic Established Populations for Epidemiologic Studies of the Elderly (H-EPESE) (1,483 subjects). The overall prevalence of falls across the seven SABE cities and the H-EPESE ranged from 21.6% in Bridgetown, Barbados, to 34.0% in Santiago, Chile. In multiple logistic regression analyses, female gender, increased age, high depressive symptoms, and having any functional limitations were significant independent risk factors for falls in most of the cities studied as well as among the elderly Mexican-Americans. In several of the cities, significant risk factors also included diabetes, urinary incontinence, and arthritis. The prevalence of falls had a large variation among the countries studied. Some of the risk factors that we identified could be modified so as to help prevent falls in older people in these populations. The factors deserving attention include depressive symptoms, functional limitations, diabetes, and urinary incontinence.


Utilization of agro-industrial waste for β-galactosidase production under solid state fermentation using halotolerant Aspergillus tubingensis GR1 isolate.
A halotolerant fungal isolate Aspergillus tubingensis GR1 was isolated from the man-made solar saltern located at Khambhat, Gujarat, India, and identified using 28S rDNA partial genome sequencing. This isolate was studied for β-galactosidase production under solid state fermentation using wheat bran and deproteinized acid cheese whey. The influence of various agro-industrial wastes, nitrogen source and other growth conditions on β-galactosidase production was investigated using 'one-factor-at-a-time' approach. Among various variables screened along with wheat bran and deproteinized acid cheese whey as major growth substrate, corn steep liquor and MgSO4 were found to be most significant. The optimum concentrations of these significant parameters were determined employing the response surface central composite design, revealing corn steep liquor concentration (2 mL) and magnesium sulphate (50 mg) per 5 g of wheat bran and 20 mL of deproteinized acid cheese whey for highest enzyme production (15,936 U/gds). These results suggest the feasibility of industrial large-scale production of β-galactosidase known to be valuable in whey hydrolysis and removal of galactosyl residue from polysaccharide.