Saving the zone of stasis: is glutathione effective?
One of the main subjects that burn researches are focused on is saving the zone of stasis. There are many molecules that are used for this purpose, but all have their drawbacks. Glutathione is one of the major buffer molecules of the cells and is known to increase the thermo-resistance of the cells. In this study, the effect of the systemic glutathione on the zone of stasis was evaluated. The results showed that glutathione is an effective molecule for saving the zone of stasis. It is well-known cheap, and easy to use.


Binaural pre-processing for contralateral sound field attenuation can improve speech-in-noise intelligibility for bilateral hearing-aid users.
We have recently proposed a binaural sound pre-processing method to attenuate sounds contralateral to each ear and shown that it can improve speech intelligibility for normal-hearing (NH) people in simulated "cocktail party" listening situations (Lopez-Poveda et al., 2022, Hear Res 418:108,469). The aim here was to evaluate if this benefit remains for hearing-impaired listeners when the method is combined with two independently functioning hearing aids, one per ear. Twelve volunteers participated in the experiments; five of them had bilateral sensorineural hearing loss and seven were NH listeners with simulated bilateral conductive hearing loss. Speech reception thresholds (SRTs) for sentences in competition with a source of steady, speech-shaped noise were measured in unilateral and bilateral listening, and for (target, masker) azimuthal angles of (0°, 0°), (270°, 45°), and (270°, 90°). Stimuli were processed through a pair of software-based multichannel, fast-acting, wide dynamic range compressors, with and without binaural pre-processing. For spatially collocated target and masker sources at 0° azimuth, the pre-processing did not affect SRTs. For spatially separated target and masker sources, the pre-processing improved SRTs when listening bilaterally (improvements up to 10.7 dB) or unilaterally with the acoustically better ear (improvements up to 13.9 dB), while it worsened SRTs when listening unilaterally with the acoustically worse ear (decrements of up to 17.0 dB). Results show that binaural pre-processing for contralateral sound attenuation can improve speech-in-noise intelligibility in laboratory tests also for bilateral hearing-aid users.


Phosphorylation of human sperm protamines HP1 and HP2: identification of phosphorylation sites.
Human sperm is characterized by a high heterogeneity of its basic nuclear protein complement of pro-protamines, protamines and histones. This heterogeneity is increased by the persistence of phosphorylated protamines in mature spermatozoa. Alkaline phosphatase treatment of whole protein indicated that protamines HP1 and HP2 were phosphorylated to various degrees. Presence of non-phosphorylated and phosphorylated protamines HP1 and HP2 was further demonstrated by electrospray mass spectrometry. Phosphorylation sites of mono- and di-phosphorylated protamine HP1 were identified by automatic Edman degradation of the protein after phosphoserine derivatization to S-ethylcysteine. In both phosphorylated forms, Ser-10 was found phosphorylated; in the di-phosphorylated form, Ser-8 was identified as the second site of phosphorylation. In protamine HP2, the unique site of phosphorylation (Ser-14) was located after limited acid hydrolysis of enzymic peptides and thin-layer electrophoresis.


Influence of patterns of neonatal transfers on breastfeeding outcomes.
To investigate/evaluate the impact of hospital transfers on breastfeeding outcomes for infants requiring neonatal intensive care and to explore approaches to supporting lactation and breastfeeding. A national study of two cohorts of infants admitted to the nine neonatal intensive care units (NICUs) in New South Wales (NSW) Australia using data sourced from a prospectively collected NICUS data collection. A total of 7829 infants were admitted to the nine NICUs in NSW for two 18-month cohorts (2012/2013 and 2015/2016) and were included in the study. These infants experienced 17 238 transfers to 107 different hospitals during the study period. Multiple transfers and low gestational age were independently associated with lower breastfeeding rates at hospital discharge across both cohorts. Extremely preterm infants and those with the greatest number of hospital transfers are least likely to be breastfed at time of discharge. Breastfeeding rates remained unchanged over time. The level of lactation support varied across NICUs. This study highlights the challenges of establishing and maintaining breastfeeding for infants requiring intensive care who have had multiple hospital transfers for continuing or specialist care. There is wide variation in lactation support across facilities. The importance of a consistent approach, education programmes and the provision of appropriate lactation support is emphasised.


Role of the beta subunit in interaction of the eighth component of human complement with the membrane-bound cytolytic complex.
This study is concerned with the subunits of the eighth component of human complement (C8) and their role in facilitating specific incorporation of C8 into the membrane-bound cytolytic complex of complement. The noncovalently associated alpha-gamma and beta subunits of C8 were purified and examined for their ability to interact independently with the C8 binding site on C5b-7, the membrane-bound precursor of the cytolytic complex. Using erythrocyte-bound C5b-7 (EAC1-7), it was observed that native C8 and its isolated beta subunit have similar high affinities for this complex. Binding of beta to EAC1-7 was specific for the C8 binding site as evidenced by the fact that 1) nearly identical molar amounts of either C8 or beta were required to saturate C8 binding sites on EAC1-7; 2) pretreatment of EAC1-7 with saturating amounts of beta prevented binding of C8; and 3) pretreatment of EAC1-7 with saturating amounts of C8 prevented binding of beta. In related experiments, alpha-gamma alone had no affinity for EAC1-7, however, binding comparable to that exhibited by native C8 occurred if alpha-gamma was first incubated with an equimolar amount of beta. Other experiments showed that if EAC1-7 was first saturated with beta to produce EAC1-7(beta), alpha-gamma also bound to yield functional C8 on the cell surface. These results provide conclusive evidence that structural features of C8 which are essential for specific recognition by membrane-bound C5b-7 are contained in the beta subunit.


Factors associated with malnutrition in children < 5 years in western Kenya: a hospital-based unmatched case control study.
Globally, under-nutrition accounts for > 3 million deaths annually among children < 5 years, with Kenya having ~ 35,000 deaths. This study aimed to identify factors associated with malnutrition in children aged < 5 years in western Kenya. We conducted a hospital-based unmatched case-control study between May and June 2017. Cases were defined as children aged 6-59 months with either z-score for weight-for-height ≤ -2SD or ≥ +2SD; weight-for-age ≤ -2SD or ≥ +2SD; or height-for-age ≤ -2SD. Controls were children aged 6-59 months with age-appropriate anthropometric measurements. Cases were consecutively recruited while systematic random sampling was used to select controls. Data from interviews and clinical records were collected and entered into Epi-Info, which was used to run unconditional logistic regression analyses. A total of 94 cases and 281 controls were recruited. Of the cases, 84% (79/94) were under-nourished. Mother not having attended ante-natal clinic (OR = 7.9; 95% CI: 1.5-41.2), deworming (OR = 0.8; 95% CI: 0.4-1.2), and pre-lacteal feeding (OR = 1.8; 95% CI: 1.1-3.0) were associated with under-nutrition. Delayed developmental milestones (AOR = 13.9; 95% CI: 2.8-68.6); low birth weight (AOR = 3.3; 95% CI: 1.4-7.6), and paternal lack of formal education (AOR = 4.9; 95% CI: 1.3-18.9) were independently associated with under-nutrition. Proper pre-natal care, child feeding practices and deworming programs should be enhanced to reduce pediatric malnutrition.


[Abdominoplasty complications: particularities of the post-bariatric patients within a 238 patients series].
Obesity is one of the health issues that is recently worldwide increasing, as well as in France. Variety of procedures of bariatric surgery have been developed over the past decade and are performed for morbid obesity hence. The demand of plastic surgery increased for body contouring and reshaping after dramatic weight loss, in which the abdominoplasty is considered as one of the most common procedures. The purpose of this study is to clarify the differences between the post-bariatric patients and others, and their influence on the occurrence of complications following abdominoplasty. Retrospective study involving a group of post-abdominoplasty patients collected from the same center between January 2000 and December 2010. Among 238 reviewed patients' files; 114 of them are post-bariatric patients, and 124 are non-bariatric patients. Several differences were found between these two groups of patients including the maximum BMI, BMI at the time of surgery, the rates of cardiovascular risk factors, and a longer operative time in the post-bariatric group. The complication rate in these patients was significantly higher (55.3% against 26.6%) with mainly healing problems. Major maximum weight, obesity at the time of surgery, long operative time and postoperative drainage system are all considered as risk factors, which carry high rate of complications. It appears that abdominoplasty in post-bariatric patients requires specific care because of particularities inherent in this population.


Obesity is associated with poor prognosis in cardiogenic arrest survivors receiving coronary angiography.
The role of body mass index (BMI) in clinical outcomes in patients resuscitated from cardiac arrest (CA) has recently drawn attention. We evaluated the effect of BMI on the prognosis of patients successfully resuscitated from cardiogenic arrest. This retrospective cohort study included 273 non-traumatic adult cardiogenic arrest survivors receiving coronary angiography after return of spontaneous circulation in three hospitals from January 2011 to September 2017. These patients were classified as underweight, normal-weight, overweight, and obese, based on BMI (<18.5; 18.5-24.9; 25.0-29.9; and ≥30 kg/m2, respectively). In-hospital mortality and poor neurological outcomes were compared among groups. The obese group had significantly higher rates of in-hospital mortality and poor neurological outcomes (cerebral performance scale = 3-5) than did the other groups (for underweight, normal-weight, overweight, and obese groups, in-hospital mortality rates were 38.5%, 29.8%, 39.0%, and 64.1%, respectively, p = 0.002; poor neurological outcomes were 53.9%, 43.8%, 47.0%, and 71.8%, respectively, p = 0.02). The obese group exhibited higher risks of in-hospital mortality and poor neurological outcomes than did the normal-weight group (in-hospital mortality: adjusted hazard ratio (aHR) = 5.21, 95% confidence interval (CI) 2.16-10.32, p < 0.001; poor neurological outcomes: aHR = 3.77, 95% CI 1.69-8.36, p = 0.002). Obesity was associated with higher risks of in-hospital mortality and poor neurological recovery.


Lamotrigine rescues neuronal alterations and prevents seizure-induced memory decline in an Alzheimer's disease mouse model.
Epilepsy is a comorbidity associated with Alzheimer's disease (AD), often starting many years earlier than memory decline. Investigating this association in the early pre-symptomatic stages of AD can unveil new mechanisms of the pathology as well as guide the use of antiepileptic drugs to prevent or delay hyperexcitability-related pathological effects of AD. We investigated the impact of repeated seizures on hippocampal memory and amyloid-β (Aβ) load in pre-symptomatic Tg2576 mice, a transgenic model of AD. Seizure induction caused memory deficits and an increase in oligomeric Aβ42 and fibrillary species selectively in pre-symptomatic transgenic mice, and not in their wildtype littermates. Electrophysiological patch-clamp recordings in ex vivo CA1 pyramidal neurons and immunoblots were carried out to investigate the neuronal alterations associated with the behavioral outcomes of Tg2576 mice. CA1 pyramidal neurons exhibited increased intrinsic excitability and lower hyperpolarization-activated Ih current. CA1 also displayed lower expression of the hyperpolarization-activated cyclic nucleotide-gated HCN1 subunit, a protein already identified as downregulated in the AD human proteome. The antiepileptic drug lamotrigine restored electrophysiological alterations and prevented both memory deficits and the increase in extracellular Aβ induced by seizures. Thus our study provides evidence of pre-symptomatic hippocampal neuronal alterations leading to hyperexcitability and associated with both higher susceptibility to seizures and to AD-specific seizure-induced memory impairment. Our findings also provide a basis for the use of the antiepileptic drug lamotrigine as a way to counteract acceleration of AD induced by seizures in the early phases of the pathology.


Assay of soluble guanylate cyclase activity by isocratic high-performance liquid chromatography.
A HPLC method alternative to labelled or unlabelled procedures was developed for the assay of guanylate cyclase (GC) activity. The substrate (GTP) and the product (cGMP) of the enzymatic reaction were separated in the isocratic mode on a muBondapak C18 column. The activity of GC was linearly dependent on the amount of cGMP produced in the presence of sodium nitroprusside. This approach was applied to follow the purification of GC from bovine lung and to evaluate its stability in different storage conditions.


Growth and developmental instability.
Developmental stability reflects the ability of an individual to develop a regular phenotype under given environmental and genetic conditions. Measures of developmental instability include the degree of fluctuating asymmetry and the frequency of phenodeviants. Endocrine, neural and circulatory mechanisms that control similar development of morphological characters on the two sides of the body are also involved in controlling overall development. Cross-validation studies have shown that measures of developmental instability are positively correlated with other measures of welfare such as tonic immobility in poultry. Asymmetric animals grow less rapidly than symmetric individuals. Eleven studies have investigated the relationship between growth rate and developmental instability, and the observed effect size (Pearson correlation coefficient adjusted for sample size) is -0.15. Studies of chickens have shown that asymmetry increases as a response to selection for increased growth rate. As conditions for rearing deteriorate by higher density, fluctuating asymmetry increases and growth rate decreases, both within and among farms. Fluctuating asymmetry can be considered a measure of animal welfare since larger values reflect worse environmental conditions as experienced by the individual animal itself. Since growth rate and fluctuating asymmetry are negatively correlated, we can infer that improvement of rearing conditions leading to reduced asymmetry will both benefit the producer (in terms of increased growth), but also the animals in terms of better conditions for rearing.


A genetic study of various enzyme polymorphisms in Pleurodeles waltlii (Urodele, Amphibian). III. The relationship between sex-linked peptidase-1 expression and gene-dose effects.
Erythrocyte peptidase-1 was analyzed by electrophoresis in various types of triploid Pleurodeles waltlii. Densitometric analysis of the zymograms showed (1) the existence of a gene-dose effect and (2) the presence of two Pep-1B alleles for one Pep-1A allele in heterozygous triploid females of biparental origin. Owing to the sex linkage of the PEP-1 locus (alleles Pep-1A and Pep-1B situated on the Z and W sex chromosomes, respectively), the results show that the sex genotype of these females is ZWW. In a particular line called series 103, the existence of a null allele was demonstrated. Densitometric analysis of females which were Pep-1A/Pep-10 (ZW), Pep-10/Pep-10 (WW), and Pep-1A/Pep-10/Pep-10 (ZWW) confirmed the gene-dose effect.


The influence of head injury on olfactory and gustatory function.
Head injury, particularly that resulting in brain injury, is a significant public health concern. For example, annual incidence rates of traumatic brain injury, a common consequence of head injury, range from 54 to 60 million people worldwide, including 2.2-3.6 million people whose trauma is moderate to severe. Trauma to the face and brain, including blast injuries common in modern warfare, can result in alterations in the ability to both smell and taste. In the case of smell, these include total loss of function (anosmia), decreased sensitivity (hyposmia), alterations in odor quality (dysosmia), and hallucination (phantosmia). Although taste dysfunction, i.e., altered perception of such basic taste-bud-mediated sensations as sweet, sour, bitter, salty, and savory (umami), can be similarly influenced by head trauma, the effects are typically more subtle and less studied. The present review provides an up-to-date assessment of what is known about the impact of head injury on quantitative measures of taste and smell function, including the influences of severity, type of injury, location of insults, prognosis, and approaches to therapy.


Identification of a third msa gene in Renibacterium salmoninarum and the associated virulence phenotype.
Renibacterium salmoninarum, a gram-positive diplococcobacillus, causes bacterial kidney disease, a condition that can result in extensive morbidity and mortality among stocks of fish. An immunodominant extracellular protein, called major soluble antigen (MSA), is encoded by two identical genes, msa1 and msa2. We found evidence for a third msa gene, msa3, which appears to be a duplication of msa1. Unlike msa1 and msa2, msa3 is not present in all isolates of R. salmoninarum. The presence of the msa3 locus does not affect total MSA production in culture conditions. In a challenge study, isolates possessing the msa3 locus reduced median survival in juvenile chinook salmon (Oncorhynchus tshawytscha) by an average of 34% at doses of < or =10(5) cells per fish compared to isolates lacking the msa3 locus. In contrast, no difference in survival was observed at the highest dose, 10(6) cells per fish. The phenotype associated with the msa3 locus and its nonuniform distribution may contribute to observed differences in virulence among R. salmoninarum isolates.


Conceptual Frameworks of Postoperative Recovery: A Scoping Review.
We conducted a scoping review to identify existing conceptual frameworks of postoperative recovery (PR) and assess their content. PR is increasingly recognized by providers and third-party payers as a multidimensional phenomenon. Efforts to optimize PR and reduce complications and readmissions continue to evolve through changes in care (i.e., enhanced recovery protocols) and financial incentives. Delineating all factors affecting PR using a conceptual framework should aid in the design of effective interventions. Web of Science and PubMed were queried to identify articles, between January 1980 and August of 2019, about conceptual frameworks of PR, using the search terms: "concept," "model," "framework," "recovery after surgery," "conceptual framework" "postoperative," "surgery," and "children." Articles considering PR as a concept rather than an outcome were included. Articles were examined in accordance with Walker and Avant's method for the concept analysis. Concepts identified across articles were classified as domains and subdomains of PR. The search yielded 183 unique articles; 8 met inclusion/exclusion criteria. Most articles defined PR as a period of days to weeks (n = 7) rather than days (n = 1). PR was mostly conceptualized as a process involving the patient and the health care system (n = 4) rather than the patient alone (n = 2). Physiological recovery (n = 8), activities of daily living (n = 8), pain (n = 5), cognitive/psychological recovery (n = 4), social recovery (n = 2), and patient perspective (n = 1) were the identified domains. Existing patient-reported outcome measures were used to assess most PR domains; however, definitions of domains and subdomains differed. None of the PR conceptual frameworks included were specific to children. There are few conceptual frameworks of PR in adults, and the definitions of PR differ. No framework was specific to children. Consensus on a conceptual framework of PR in adults and development of a conceptual framework of PR specific to children are needed.


New mechanism contributing to drug-induced arrhythmia: rescue of a misprocessed LQT3 mutant.
The cardiac sodium channel (SCN5A) mutation L1825P has been identified in a patient with drug-induced torsade de pointes precipitated by the IKr blocker cisapride. Although L1825P generates late sodium current typical of SCN5A-linked long-QT syndrome (LQT3) in vitro, the patient reported had a normal QT interval before administration of the drug. To address this discrepancy, we tested the hypothesis that this mutant channel is not processed normally. CHO cells transfected with L1825P displayed significantly reduced peak INa (209+/-36 versus 23+/-3 pA/pF, P<0.05). Confocal imaging and cell-counting studies using epitope-tagged constructs demonstrated that cell surface expression of the mutant was only approximately 9% of wild-type. Incubating transfected cells with cisapride partially rescued misprocessing to 30% of wild-type. As a result, "late" sodium current increased with cisapride from 1.2+/-0.11 to 5.04+/-0.77 pA/pF (P<0.05). L1825P fails to generate QT prolongation because it does not reach the cell surface. Moreover, the data suggest that cisapride caused torsade de pointes not only by blocking IKr but also by rescuing cell surface expression of the mutant channel, further exaggerating the LQT3 phenotype. This not only represents a new mechanism in the drug-induced long-QT syndrome but also strongly supports the concept that variable cell surface expression contributes to clinical variability in the LQT3 phenotype.


Effect of Intermolecular Interactions on Metal-to-Metal Charge Transfer: A Combined Experimental and Theoretical Investigation.
Understanding the effects of intermolecular interactions on metal-to-metal charge transfer (MMCT) is crucial to develop molecular devices by grafting MMCT-based molecular arrays. Herein, we report a series of solvent-free {Fe2 Co2 } compounds sharing the same cationic tetranuclear {[Fe(PzTp)(CN)3 ]2 [Co(dpq)2 ]2 }2+ (PzTp- =tetrakis(pyrazolyl)borate, dpq=dipyrido[3,2-d:2',3'-f]quinoxaline) square units but having anions with different size, including BF4 - , PF6 - , OTf- , and [Fe(PzTp)(CN)3 ]- . Intermolecular π⋅⋅⋅π interactions between dpq ligands, which coordinate to cobalt ions in the {[Fe(PzTp)(CN)3 ]2 [Co(dpq)2 ]2 }2+ units, can be modulated by introducing different counterions, regulating the distortion of the CoN6 octahedron and ligand field around the cobalt ions. This change results in different MMCT behavior. Computational analyzes reveal the substantial role of the intermolecular interactions tuned by the presence of different counteranions on the MMCT behavior.


A new solid-phase radioimmunoassay to detect anti-GAD65 autoantibodies.
This paper describes a simple, rapid, routine method to detect anti-GAD65 autoantibodies by a solid-phase radioimmunoassay using human recombinant GAD65 coated microwells and 125I-protein A to reveal antibody binding. Both recombinant and radiolabelled proteins are commercially available. This new method was validated by investigating the presence of GAD65 autoantibodies in two different studies (A and B); the first including subjects originating from our own case histories (group A sera), the second made up of recoded subjects and standards sent to our lab by the Second International GAD Antibody Workshop organizers (group B sera). In study A we tested sera from 52 normal subjects, 25 newly diagnosed type 1 diabetics and 3 stiff man syndrome (SMS) subjects detecting GAD65 autoantibodies in 72% of IDDM and 100% of SMS patients. In study B we tested (in blind fashion) 89 recoded sample sera or standards that were part of the larger group used in the Second International GAD Antibody Workshop, finding GAD65 autoantibodies in 3.3% of healthy control subjects (1/30), 60% of IDDM patients (18/30), 100% of ICA + nondiabetic subjects (3/3) but in none of 4 nondiabetic patients with Graves disease. Comparing our solid-phase RIA results with those published for the same sera from the Second International GAD Antibody Workshop we obtained for our method a sensitivity of 85.7%, a specificity of 93.9% and a consistency of 100%. These result indicate that our assay, which is based on commercially available reagents, should be a useful tool for the detection of GAD65 autoantibodies in large scale studies.


Neuron-specific enolase and S 100B protein as predictors of outcome in ischaemic stroke.
As neuron-specific enolase (NSE) and S 100B protein are brain origin proteins, the aim of this study was to assess whether a single NSE and S 100B measure may predict clinical outcome of patients with cerebral ischaemic infarct. Seventy-one patients with ischaemic stroke and 41 controls were studied. All patients had computed tomography of the brain performed after admission and on the third day and volume of the infarct was assessed by the volumetric method from the second examination. NSE and S 100B protein were analysed by immunochemiluminescence on the fourth day after admission. Clinical state of the patients was determined with the NIH stroke, Barthel and Rankin scales on admission, discharge from the hospital, and after one and 3 months from the onset of stroke. NSE levels in blood were significantly higher in stroke patients than in the control group - 36.9 ± 24.0 vs. 14.3 ± 9.7 μg/L. Also, the levels of the S 100B protein were significantly higher in the patient group (0.85 ± 1.74 vs. 0.10 ± 0.03 μg/L) but only the levels of S 100B protein correlated with the calculated size of the infarct (Spearman coefficient = 0.77). No such correlation was identified for NSE level (Spearman coefficient = 0.25). Although significant differences in NSE and S 100B levels between stroke patients and the control group were found, only S 100B protein level correlated with stroke volume, neurological status at admission and functional outcome. NSE did not correlate with stroke volume, neurological status or clinical outcome.


Anemia and Associated Factors Among Pregnant Women Attending Antenatal Care at Madda Walabu University Goba Referral Hospital, Bale Zone, Southeast Ethiopia.
Globally, anemia is a public health problem, particularly in developing countries, including Ethiopia. Pregnant women are at a higher risk for anemia and anemic pregnant women often develop complications. Early diagnosis of pregnant women for anemia saves both the life of the newborn and hers. Therefore, the present study intended to assess anemia and its associated factors among pregnant women attending at Madda Walabu University Goba referral hospital. An institutional-based cross-sectional study was conducted from April 3 to May 3, 2019, among pregnant women attending Madda Walabu University Goba referral hospital. A total of 353 respondents were included in the study by using systematic random sampling techniques. Data were collected using both face-to-face interview and hemoglobin measurement. The collected data were entered into SPSS and analyzed using both bi-variable and multiple logistic regressions. Finally, statistical significance was declared at a p-value of less than 0.05 in the multiple logistic regressions. In this study, the prevalence of anemia was 46.2% (95%CI: 40.9%, 51.5%). Factors significantly associated with anemia were positive stool examination for parasites (AOR=2.9, 95%CI 1.255-6.856), lower consumption of fruits and vegetables (AOR=2.7, 95%CI 1.47-4.86), no iron supplementation (AOR=2.2, 95%CI 1.07-4.45), large family size (AOR=2.1, 95%CI 1.13-3.77), and absence of abortion history (AOR=0.4, 95%CI 0.24-0.77). Prevalence of anemia among pregnant women was high. Pregnant women with a previous history of abortion, positive stool for parasite, lower consumption of fruits and vegetables, no iron supplementation and large family size above five were risk factors for anemia. Therefore, encouraging pregnant women to take iron supplementation and to eat fruits and vegetables are important.


[Does the healthcare for rare diseases benefit from the legislative reforms?]
The founding of the National Action League for People with Rare Diseases (NAMSE) in 2010 represents the creation of a significant political platform. In addition, recent years had seen Germany and the EU adopt specific legislative measures aimed at improving healthcare for people with rare diseases. In this article we will give an overview of the legislative reforms adopted between 2013 and 2016 and evaluate how the specific healthcare situation of people with rare diseases has been improved. This article analyzes the health care legislative reforms adopted during the 18th term (since 2013) of the German lower house, the Bundestag, as well as their self-governing implementation. The analysis also extends to similar political initiatives of the European Commission. The impact of the recent hospital reforms on the health care received by patients or on the work of health care providers in the field of rare diseases cannot be assessed conclusively at this point (January 2017). One positive feature is that the health care coverage mandate of the university hospital outpatient departments now also comprises rare diseases. Recent legislative measures have created possibilities to improve the economic position of centers for rare diseases and university hospital outpatient departments. What these improvements will look like specifically depends on the implementation within the hospital plans of the federal states as well as on the outcome of the remuneration negotiations between university hospitals and health insurance funds.


Effect of inhibitors of GABA aminotransferase on the metabolism of GABA in brain tissue and synaptosomal fractions.
Five inhibitors of the GABA degrading enzyme GABA-aminotransferase (GABA-T), viz., gabaculine, gamma-acetylenic GABA, gamma-vinyl GABA, ethanolamine O-sulphate, and aminooxyacetic acid, as well as GABA itself and the antiepileptic sodium valproate were administered to mice in doses equieffective to raise the electroconvulsive threshold by 30 V. The animals were killed at the time of maximal anticonvulsant effect of the respective drugs and GABA, GABA-T and glutamate decarboxylase (GAD) were determined in whole brain and synaptosomes, respectively. The synaptosomal fraction was prepared from brain by conventional ultracentrifugation procedures. All drugs studied brought about significant increases in both whole brain and synaptosomal GABA concentrations, and, except GABA itself, inhibited the activity of GABA-T. Furthermore, all drugs, except GABA and gamma-acetylenic GABA, activated GAD in the synaptosomal fraction. This was most pronounced with ethanolamine O-sulphate, which induced a twofold activation of this enzyme but exerted only a weak inhibitory effect on GABA-T. The results suggest that activation of GAD is an important factor in the mechanism by which several inhibitors of GABA-T and also valproate increase GABA concentrations in nerve terminals, at least in the relatively non-toxic doses as used in this study.


Malnutrition prevalence and associated biochemical factors among drug-resistance tuberculosis (DR-TB) patients at key treatment sites in Conakry City, Republic of Guinea.
drug-resistant tuberculosis is a major global health problem and a threat to health security given the increase in the number of cases and the challenges associated with care. Besides, the relationship between poor nutritional status and tuberculosis is clearly established. For relevant and evidence-based public health decision-making regarding the management of malnutrition in patients with drug-resistant tuberculosis in the initial phase, it is essential to estimate the prevalence of malnutrition and understand the risk factors associated with it. we performed a retrospective cohort study in drug-resistant tuberculosis patients aged 18 years and older, among which the nutritional status was assessed through BMI. All predictors were included in a prediction model using the multivariate logistic model according to the lowest Akaike criterion. Discrimination and model calibration was evaluated using receiver performance analysis, and the Hosmer and Lemeshow test. this study revealed a prevalence of malnutrition of 64.7% in drug-resistant tuberculosis patients in our 218-patient series. The factors associated with malnutrition were: unsuccessful treatment, the active presence of mycobacterium tuberculosis, increased bacteriological conversion time, increased serum creatinine, increased transaminase SGPT of the liver, and anaemia. Some of the factors not associated with malnutrition included the history of anti-tuberculosis treatment, vomiting, hepatic SGPT, initial AFB count, smear and culture conversion time, depression, and chest X-ray. malnutrition remains a concern among drug-resistant tuberculosis patients in Guinea as it affects more than half of them with a negative impact on the outcome of treatment. Implementing specific interventions for these high-risk patients, including nutritional supplementation, psychosocial support, and treatment for tuberculosis, can improve management for better treatment outcomes.


Boerhaave's syndrome: an elusive diagnosis.
Boerhaave's syndrome represents a diagnostic dilemma for the emergency physician. The prognosis of this truly life-threatening emergency is darkened by any significant diagnostic delay. Unfortunately, classic or expected symptoms and signs are frequently absent at presentation, a circumstance that leads to frequent misdiagnosis. Two cases of Boerhaave's syndrome with "atypical" clinical presentations are reviewed and discussed. It is clear that Boerhaave's syndrome should always be suspected in the evaluation of any sudden chest, abdominal, or back pain associated with emesis. However, emphasis should be placed on the fact that this entity may occur without emesis. The chest radiograph is the most helpful diagnostic aid. Undoubtedly, maintenance of a high degree of suspicion by the emergency physician for Boerhaave's syndrome will lead consistently to earlier diagnosis, and subsequent aggressive intervention should result in considerable reduction in rates of both morbidity and mortality.


Epitope Mapping Using pEX.
Monoclonal and polyclonal antibodies are important tools in molecular and cell biology, allowing protein molecules to be purified (1), identified in immunoblots of gels (2) and sections of biological material (3), and probed for functionally active regions. Studies on the nature of antibody binding sites show that the molecular interaction between an antibody and its antigen is very specific and localized to a small patch on the surface of a molecule (4). For antibodies that inhibit molecular function, it is clearly of value to be able to map their binding site to a physical location on the polypeptide chain. Even antibodies that do not inhibit binding or catalysis of the protein can give useful information since the epitopes of antibodies are located on the surface of an antigen molecule (5-8). Two classes of antibody binding site have been distinguished. In the first-"continuous" epitopes-synthetic peptides are capable of binding to the antibody (9), suggesting that a single loop of the polypeptide chain is predominantly responsible for the antibody binding. In the second-discontinuous epitopes-linear peptides do not bind to the antibody, but the important amino acids can be mapped by mutational analysis (10). These discontinuous epitopes can sometimes be mimicked by peptides that imitate the spacial orientation of the surface residues (11).


Deletion of Nf2 in neural crest-derived tongue mesenchyme alters tongue shape and size, Hippo signalling and cell proliferation in a region- and stage-specific manner.
The mammalian tongue develops from the branchial arches (1-4) and comprises highly organized tissues compartmentalized by mesenchyme/connective tissue that is largely derived from neural crest (NC). This study aimed to understand the roles of tumour suppressor Neurofibromin 2 (Nf2) in NC-derived tongue mesenchyme in regulating Hippo signalling and cell proliferation for the proper development of tongue shape and size. Conditional knockout (cKO) of Nf2 in NC cell lineage was generated using Wnt1-Cre (Wnt1-Cre/Nf2cKO ). Nf2 expression, Hippo signalling activities, cell proliferation and tongue shape and size were thoroughly analysed in different tongue regions and tissue types of Wnt1-Cre/Nf2cKO and Cre- /Nf2fx/fx littermates at various stages (E10.5-E18.5). In contrast to many other organs in which the Nf2/Hippo pathway activity restrains growth and cell proliferation and as a result, loss of Nf2 decreases Hippo pathway activity and promotes an enlarged organ development, here we report our observations of distinct, tongue region- and stage-specific alterations of Hippo signalling activity and cell proliferation in Nf2cKO in NC-derived tongue mesenchyme. Compared to Cre- /Nf2fx / fx littermates, Wnt1-Cre/Nf2cKO depicted a non-proportionally enlarged tongue (macroglossia) at E12.5-E13.5 and microglossia at later stages (E15.5-E18.5). Specifically, at E12.5 Nf2cKO mutants had a decreased level of Hippo signalling transcription factor Yes-associated protein (Yap), Yap target genes and cell proliferation anteriorly, while having an increased Yap, Yap target genes and cell proliferation posteriorly, which lead to a tip-pointed and posteriorly widened tongue. At E15.5, loss of Nf2 in the NC lineage resulted in distinct changes in cell proliferation in different regions, that is, high in epithelium and mesenchyme subjacent to the epithelium, and lower in deeper layers of the mesenchyme. At E18.5, cell proliferation was reduced throughout the Nf2cKO tongue.


Large-scale surgical workflow segmentation for laparoscopic sacrocolpopexy.
Laparoscopic sacrocolpopexy is the gold standard procedure for the management of vaginal vault prolapse. Studying surgical skills and different approaches to this procedure requires an analysis at the level of each of its individual phases, thus motivating investigation of automated surgical workflow for expediting this research. Phase durations in this procedure are significantly larger and more variable than commonly available benchmarks such as Cholec80, and we assess these differences. We introduce sequence-to-sequence (seq2seq) models for coarse-level phase segmentation in order to deal with highly variable phase durations in Sacrocolpopexy. Multiple architectures (LSTM and transformer), configurations (time-shifted, time-synchronous), and training strategies are tested with this novel framework to explore its flexibility. We perform 7-fold cross-validation on a dataset with 14 complete videos of sacrocolpopexy. We perform both a frame-based (accuracy, F1-score) and an event-based (Ward metric) evaluation of our algorithms and show that different architectures present a trade-off between higher number of accurate frames (LSTM, Mode average) or more consistent ordering of phase transitions (Transformer). We compare the implementations on the widely used Cholec80 dataset and verify that relative performances are different to those in Sacrocolpopexy. We show that workflow segmentation of Sacrocolpopexy videos has specific challenges that are different to the widely used benchmark Cholec80 and require dedicated approaches to deal with the significantly larger phase durations. We demonstrate the feasibility of seq2seq models in Sacrocolpopexy, a broad framework that can be further explored with new configurations. We show that an event-based evaluation metric is useful to evaluate workflow segmentation algorithms and provides complementary insight to the more commonly used metrics such as accuracy or F1-score.


Sulphate-climate coupling over the past 300,000 years in inland Antarctica.
Sulphate aerosols, particularly micrometre-sized particles of sulphate salt and sulphate-adhered dust, can act as cloud condensation nuclei, leading to increased solar scattering that cools Earth's climate. Evidence for such a coupling may lie in the sulphate record from polar ice cores, but previous analyses of melted ice-core samples have provided only sulphate ion concentrations, which may be due to sulphuric acid. Here we present profiles of sulphate salt and sulphate-adhered dust fluxes over the past 300,000 years from the Dome Fuji ice core in inland Antarctica. Our results show a nearly constant flux of sulphate-adhered dust through glacial and interglacial periods despite the large increases in total dust flux during glacial maxima. The sulphate salt flux, however, correlates inversely with temperature, suggesting a climatic coupling between particulate sulphur and temperature. For example, the total sulphate salt flux during the Last Glacial Maximum averages 5.78 mg m(-2) yr(-1), which is almost twice the Holocene value. Although it is based on a modern analogue with considerable uncertainties when applied to the ice-core record, this analysis indicates that the glacial-to-interglacial decrease in sulphate would lessen the aerosol indirect effects on cloud lifetime and albedo, leading to an Antarctic warming of 0.1 to 5 kelvin.


Determination of similarity measures for pairs of mass lesions on mammograms by use of BI-RADS lesion descriptors and image features.
To determine similarity measures for selection of pathology-known similar images that would be useful for radiologists as a reference guide in the diagnosis of new breast lesions on mammograms. The images were obtained from the Digital Database for Screening Mammography developed by the University of South Florida. For determination and evaluation of similarity measures, the "gold standard" of similarities for 300 pairs of masses was determined by 10 breast radiologists. For determining similarity measures that would agree with radiologists' similarity determination, an artificial neural network (ANN) was trained with the radiologists' subjective similarity ratings and the image features. The image features were determined subjectively using the Breast Imaging Reporting and Data System (BI-RADS) lesion descriptors and objectively by computerized image analysis. The similarity measures determined by the ANN were compared to the gold standard and evaluated in terms of the correlation coefficient. The similarity measures determined using the BI-RADS descriptors only were not as useful as those determined by use of the image features only. When the BI-RADS margin ratings were combined with the image features, the correlation coefficient between the subjective ratings and the objective measures improved slightly (r = 0.76) compared to those based on the image features alone (r = 0.74). The inclusion of the BI-RADS margin descriptors may be useful for determination of similarity measures, especially when it is difficult to obtain the manual outlines of the masses and if the BI-RADS descriptors were provided consistently by radiologists.


Therapeutic advances of miRNAs: A preclinical and clinical update.
miRNAs, a class of small endogenous RNAs, are one of the essential biopharmaceuticals which are in commercial spans as next-generation medicine in recent times. A snapshot of the current scenario regarding the miRNAs as biopharmaceuticals have been discussed. In this work, biopharmaceutical companies working with miRNAs and the current status of preclinical/clinical trials about miRNA therapeutics have been reviewed. Finally, recent updates on the absorption, distribution, metabolism, and excretion (ADME), as well as a delivery system of miRNAs, have been illustrated.


Kinetic modelling of ultrasound-assisted extraction of phenolics from cereal brans.
Cereal brans are by-products of the milling of cereal grains, which are mainly used as low value ingredients in animal feed. Wheat and oat bran is a rich source of bioactives and phytochemicals, especially phenolic compounds. Within this study, the application of ultrasound (US) technology to assist the extraction of phenolics from oat and wheat bran was investigated (20-45 kHz). Peleg's mathematical model was used to study the kinetics of ultrasound-assisted extraction (UAE) and subsequent stirring of total phenolic compounds (TPC). The surface morphology of cereal brans after extraction was studied using SEM analysis. The excellent agreement was determined between the values of TPC calculated from Peleg's mathematical model and actual experimental results. The constant that represents a time required for the initial phenolic concentration to be extracted to one-half of its initial value has been introduced (K1/2). It was shown that the TPC extraction kinetics was dependent only on K1/2 enabling fast kinetics fitting and comparison between extraction rates. Moreover, different values of K1/2 constant could indicate the differences in brans composition and consequently different influence of US pretreatment on these samples.


3-[2,4-Dimethoxybenzylidene]anabaseine (DMXB) selectively activates rat alpha7 receptors and improves memory-related behaviors in a mecamylamine-sensitive manner.
The alpha7 nicotinic receptor agonist 3-[2,4-dimethoxybenzylidene]anabaseine (DMXB; GTS-21) was investigated for its ability to: (1) activate a variety of nicotinic receptor subtypes in Xenopus oocytes; (2) improve passive avoidance and spatial Morris water task performances in mecamylamine-sensitive manners in bilaterally nucleus basalis lesioned rats; and (3) elevate high-affinity [3H]acetylcholine (ACh) and high-affinity alpha-[125I]bungarotoxin binding in rat neocortex following 2 weeks of daily injections. DMXB (100 microM) activated alpha7 homo-oligomeric receptors, without significant activity at alpha2-, alpha3- and alpha4-containing subtypes. Mecamylamine blocked rat alpha7 receptors weakly if co-administered with agonist, but much more potently when pre-applied. Bilateral ibotenic acid lesions of the nucleus basalis interfered with passive avoidance and spatial memory-related behaviors. DMXB (0.5 mg/kg, i.p.) improved passive avoidance behavior in lesioned animals in a mecamylamine-sensitive manner. DMXB (0.5 mg/kg 15 min before each session) also improved performance in the training and probe components of the Morris water task. DMXB-induced improvement in the probe component but not the training phase was mecamylamine-sensitive. [3H]ACh binding was elevated after 14 days of daily i.p. injections with 0.2 mg/kg nicotine but not after 1 mg/kg DMXB. Neither drug elevated high-affinity alpha-[125I]bungarorotoxin binding over this interval.


Distance between laser head and skin does not influence skin blood flow values recorded by laser speckle imaging.
Laser Speckle contrast imaging (LSCI) allows non-contact, real-time recording of cutaneous blood flow (CBF). Different distances from laser-head to skin (distances(L-S)) can be chosen by the operator to perform these recordings. We aimed to evaluate the impact of different Distances(L-S) on the analysis of rest blood flow and post-occlusive reactive hyperemia (PORH). Four distances(L-S) (10, 15, 20, and 30 cm) were evaluated in a random order in 11 healthy subjects. We analyzed the concordance between each recording at each distance(L-S). We compared CBF results (absolute values and cutaneous vascular conductance (CBF divided by mean arterial pressure)) obtained for each distance(L-S). The intra-subject coefficients of variation due to distances(L-S) (intra-CV, in%) were also studied. The mean "r" (standard deviation) cross-correlation coefficient was 0.99 (0.00) between each CBF trace issued from different distance(L-S). Both kinds of CBF results, at rest and for PORH peak, show non-significant differences when the distance(L-S) is modified. The intra-CV varies from 5.9% to 8.6% at rest and from 5.6% to 9.1% for the PORH peak. Distance(L-S) neither influences SBFR at rest, nor at peak post-occlusive hyperemia in the 10-30 cm interval using LSCI.


Predictive value of pre-arrest albumin level with GO-FAR score in patients with in-hospital cardiac arrest.
We investigated whether combining the pre-arrest serum albumin level could improve the performance of the Good Outcome Following Attempted Resuscitation (GO-FAR) score for predicting neurologic outcomes in in-hospital cardiac arrest patients. Adult patients who were admitted to a tertiary care hospital between 2013 and 2017 were assessed. Their pre-arrest serum albumin levels were measured within 24 h before the cardiac arrest. According to albumin levels, the patients were divided into quartiles and were assigned 1, 0, 0, and, - 2 points. Patients were allocated to the derivation (n = 419) and validation (n = 444) cohorts. The proportion of favorable outcome increased in a stepwise manner across increasing quartiles (p for trend < 0.018). Area under receiver operating characteristic curve (AUROC) of the albumin-added model was significantly higher than that of the original GO-FAR model (0.848 vs. 0.839; p = 0.033). The results were consistent in the validation cohort (AUROC 0.799 vs. 0.791; p = 0.034). Net reclassification indices of the albumin-added model were 0.059 (95% confidence interval [CI] - 0.037 to 0.094) and 0.072 (95% CI 0.013-0.132) in the derivation and validation cohorts, respectively. An improvement in predictive performance was found by adding the ordinal scale of pre-arrest albumin levels to the original GO-FAR score.


Needs and demands for psychological care in university students.
The interest of universities in promoting healthy habits and offering psychological assistance and treatment is the result of a desire to improve the quality of students' education and day-to-day lives. An analysis of students' needs and of the demand for psychological care is a prerequisite for implementing effective interventions. A total of 706 second-year undergraduate students (MDage = 20.2; SD = 2.73) from all branches of knowledge of the University of Oviedo (Spain) were evaluated by means of standardized instruments. Results showed academic conditions to be the principal stressors and no distinctive behaviour profiles were found according to branch of studies; 44.7% showed levels of emotional distress indicative of anxiety and 13.5% of depression. The situations generating the greatest demand for psychological assistance were fear of speaking in public, requesting help regarding personal problems, distancing oneself from worries and emotions, and the promotion of healthy sleeping and eating habits. The prevention and treatment of psychological problems has consistently been shown to be a factor which enhances academic performance. The needs identified, shared by other university populations, underline the importance of providing specific psychological care in the university context.


Ethyl pyruvate improves white matter remodeling in rats after traumatic brain injury.
Severe traumatic brain injury (TBI) results in long-term neurological deficits associated with white matter injury (WMI). Ethyl pyruvate (EP) is a simple derivative of the endogenous energy substrate pyruvate with neuroprotective properties, but its role in recovery from WMI has not been explored. This study examines the effect of EP treatment on rats following TBI using behavioral tests and white matter histological analysis up to 28 days post-injury. Anaesthetised adult rats were subjected to TBI by controlled cortical impact. After surgery, EP or Ringers solution (RS) was administrated intraperitoneally at 15 min after TBI and again at 12, 24, 36, 48, and 60 h after TBI. Sensorimotor deficits were evaluated up to day 21 after TBI by four independent tests. Immunofluorescence and transmission electron microscopy (TEM) were performed to assess white matter injury. Microglia activation and related inflammatory molecules were examined up to day 14 after TBI by immunohistochemistry or real-time PCR. Here, we demonstrate that EP improves sensorimotor function following TBI as well as improves white matter outcomes up to 28 d after TBI, as shown by reduced myelin loss. Furthermore, EP administration during the acute phase of TBI recovery shifted microglia polarization toward the anti-inflammatoryM2 phenotype, modulating the release of inflammatory-related factors. EP treatment may protect TBI-induced WMI via modulating microglia polarization toward M2.


Barley transcription factor HvNLP2 mediates nitrate signaling and affects nitrogen use efficiency.
Plants have evolved complex mechanisms to adapt to the changing nitrogen levels in the environment. In Arabidopsis, more than a dozen nitrate signaling regulatory genes have been characterized, including the NODULE INCEPTION-LIKE PROTEIN (AtNLP) genes, which play essential roles in nitrate signaling. However, whether NLP genes in the Triticeae crops are involved in nitrate regulation and nitrogen use efficiency (NUE) remains unknown. Here, we isolated a barley (Hordeum vulgare L.) mutant, hvnlp2-1, from a TILLING (Targeting Local Lesions IN Genomes) population and constructed two RNAi lines, hvnlp2-2 and hvnlp2-3, to study the function of HvNLP2. The expression of the nitrate-responsive genes was substantially inhibited after nitrate treatment in the hvnlp2 mutants, indicating that HvNLP2 controls nitrate signaling. Nitrate content was significantly higher in the hvnlp2 mutants, which may result from the decreased assimilation of nitrogen caused by reduced nitrate reductase activity and expression of nitrate assimilatory genes. HvNLP2 is localized to the nucleus in the presence of nitrate. Further investigation showed that HvNLP2 binds to and activates the nitrate-responsive cis-elements. Moreover, hvnlp2 exhibited reduced biomass, seed yield, and NUE. Therefore, HvNLP2 controls nitrate signaling and plays an important role in NUE.


Dissolved oxygen amperometric sensor based on layer-by-layer assembly using host-guest supramolecular interactions.
The development of a simple, efficient and sensitive sensor for dissolved oxygen is proposed using the host-guest binding of a supramolecular complex at a host surface by combining a self-assembled monolayer (SAM) of mono-(6-deoxy-6-mercapto)-beta-cyclodextrin (betaCDSH), iron (III) tetra-(N-methyl-4-pyridyl)-porphyrin (FeTMPyP) and cyclodextrin-functionalized gold nanoparticles (CDAuNP). The supramolecular modified electrode showed excellent catalytic activity for oxygen reduction. The reduction potential of oxygen was shifted about 200 mV toward less negative values with this modified electrode, presenting a peak current much higher than those observed on a bare gold electrode. Cyclic voltammetry and rotating disk electrode (RDE) experiments indicated that the oxygen reduction reaction involves probably 4-electrons with a rate constant (k(obs)) of 7 x 10(4) mol(-1) Ls(-1). A linear response range from 0.2 up to 6.5 mg L(-1), with a sensitivity of 5.5 microA L mg(-1) (or 77.5 microA cm(-2) L mg(-1)) and a detection limit of 0.02 mg L(-1) was obtained with this sensor. The repeatability of the proposed sensor, evaluated in terms of relative standard deviation was 3.0% for 10 measurements of a solution of 6.5 mg L(-1) oxygen.


Kisspeptin drives germ cell progression in the anuran amphibian Pelophylax esculentus: a study carried out in ex vivo testes.
Kisspeptin, via Gpr54 receptor, regulates puberty onset in most vertebrates. Thus, the direct involvement of kisspeptin activity in testis physiology was investigated in the anuran amphibian, Pelophylax esculentus. In this vertebrate gpr54 mRNA has been localized in both interstitial compartment and spermatogonia (SPG), whereas SPG proliferation requires the cooperation between estradiol and testicular Gonadotropin releasing hormone (Gnrh). In the pre-reproductive period, dose response curve to assess the effects of Kisspeptin-10 (Kp-10) was carried out in vitro (dose range: 10(-9)-10(-6)M; incubation times: 1 and 4h); proliferative activity and germ cell progression were evaluated by expression analysis of proliferating cell nuclear antigen (pcna), estrogen receptor beta (erβ), Gnrh system (gnrh1, gnrh2, gnrhr1, r2, r3) and by the count of empty, mitotic and meiotic tubules. All selected markers were up regulated at 4h Kp-10 incubation. Histological analysis also proved the increase of mitotic activity and the progression of spermatogenesis. Besides Kp-10 modulation of testicular Gnrh system, in vitro treatment with 17β-estradiol (10(-6)M) ± the antagonist ICI182-780 (10(-5)M) revealed gnrh2 and gnrhr3 estrogen dependent expression. In the reproductive period, testes were incubated for 1 and 4h with Kp-10 (10(-7)M) or Kp-10 (10(-7)M)+kisspeptin antagonist [Kp-234 (10(-6)M)]. Results obtained in the pre-reproductive period were confirmed and Kp-234 completely counteracted Kp-10 effects. In conclusion, Kp-10 modulated the expression of pcna, erβ, gnrhs and gnrhrs, inducing the progression of the spermatogenesis.


Aryl radical-mediated N-heterocyclic carbene catalysis.
There have been significant advancements in radical reactions using organocatalysts in modern organic synthesis. Recently, NHC-catalyzed radical reactions initiated by single electron transfer processes have been actively studied. However, the reported examples have been limited to catalysis mediated by alkyl radicals. In this article, the NHC organocatalysis mediated by aryl radicals has been achieved. The enolate form of the Breslow intermediate derived from an aldehyde and thiazolium-type NHC in the presence of a base undergoes single electron transfer to an aryl iodide, providing an aryl radical. The catalytically generated aryl radical could be exploited as an arylating reagent for radical relay-type arylacylation of styrenes and as a hydrogen atom abstraction reagent for α-amino C(sp3)-H acylation of secondary amides.


Dysgerminoma of the ovary: radiation therapy for recurrence and metastases.
Nine cases of dysgerminoma are reported. Five patients developed recurrence of disease in either the pelvis or metastatic sites including the lung and brain. The metastatic disease was controlled by a radiation dose of 2,000 to 3,000 rads during three to four weeks in four of the patients. The excellent response of dysgerminoma to radiation warrants an optimistic prognosis and vigilant follow-up since metastatic disease can be controlled by moderate radiation doses.


Silicon beneficial effects on yield, fruit quality and shelf-life of strawberries grown in different culture substrates under different iron status.
The silicon application either as foliar or to the radicular system of strawberry plants was investigated. Fortuna strawberry plants were grown in two different substrates (coconut fibre and organic substrate) under optimal (20 μM) or low (5 μM) iron (Fe) conditions. During the study, crop parameters including leaf area, SPAD and fruit yield were measured. At harvest, fruit quality and post-harvest shelf-life were evaluated. Results indicated that "Fortuna" strawberries plants had a poor development in coconut fibre and excellent growth and yield in the organic substrate. In the coconut fibre substrate, no differences in foliar area, fruit diameter, colour, pH and shelf-life were observed related to the Si addition under deficient Fe conditions, but an increased in weight and the firmness of the fruits, as well as in fructose content was shown. However, when 20 μM Fe were supplied, the root application of Si significantly increases: protein, mineral and sugar content, as well as fruit shelf-life by an average of 1.5 days. Likewise, the radicular silicon application to the organic substrate considerably improved yield, fruit diameter, fruit weight, glucose and fructose fruit content and the fruit shelf-life without causing distinguishable chemical or physicochemical changes. In summary, Si application to Fortuna strawberries through the roots could be a good solution to increase fruit quality and yield and to increase benefits from the agronomical point of view. Further studies in other strawberry varieties and dose rates will allow knowing with better precision how the radicular application of silicon contributes to yield and fruit shelf-life.


Sexual behavior: stimulation by painful electrical shock to skin in male rats.
A mildly painful electric shock lasting 0.5 second was delivered every 30 seconds to the skin of male rats while they were with a receptive female rat. Sexual behavior occurred, with brief latency, after shocks; thus, successive shocks resulted in a pacing of sexual behavior. This effect is attributed to periodic augmentation of sexual arousal by a periodic, nonspecific arousing stimulus (that is, shock).


Incubation period of wild type of SARS-CoV-2 infections by age, gender, and epidemic periods.
The incubation period of the coronavirus disease 2019 (COVID-19) is estimated to vary by demographic factors and the COVID-19 epidemic periods. This study examined the incubation period of the wild type of SARS-CoV-2 infections by the different age groups, gender, and epidemic periods in South Korea. We collected COVID-19 patient data from the Korean public health authorities and estimated the incubation period by fitting three different distributions, including log-normal, gamma, and Weibull distributions, after stratification by gender and age groups. To identify any temporal impact on the incubation period, we divided the study period into two different epidemic periods (Period-1: 19 January-19 April 2020 and Period-2: 20 April-16 October 2020), and assessed for any differences. We identified the log-normal as the best-fit model. The estimated median incubation period was 4.6 (95% CI: 3.9-4.9) days, and the 95th percentile was 11.7 (95% CI: 10.2-12.2) days. We found that the incubation period did not differ significantly between males and females (p = 0.42), age groups (p = 0.60), and the two different epidemic periods (p = 0.77). The incubation period of wild type of SARS-CoV-2 infection during the COVID-19 pandemic 2020, in South Korea, does not likely differ by age group, gender and epidemic period.


Cytosolic calcium responses induced by photolytic release of 1,4,5-inositol trisphosphate in single human fibroblasts.
We have used the whole cell technique to microinject human fibroblasts with either 1,4,5-inositol trisphosphate (InsP3) or 'caged' InsP3, in order to study the mechanisms of transmembrane signalling related to mitogenic stimulations. Cytosolic Ca2+ elevations in response to 1,4,5 InsP3 diffusing from the patch pipette were difficult to detect, while 1,4,5 InsP3, photoreleased after loading the cell with its inactive precursor, was capable of generating not only a single cytosolic Ca2+ rise but sometimes triggered an oscillatory calcium response, similar to that often observed under mitogenic stimulation. We estimated that less than 100 nM InsP3 was sufficient to generate Ca2+ responses. The Ca2+ rise produced by the photoreleased InsP3 could fully activate the K+ channels present in the plasma membrane of human fibroblasts.


Randomized clinical trial of nitazoxanide or sofosbuvir/daclatasvir for the prevention of SARS-CoV-2 infection.
The COVER trial evaluated whether nitazoxanide or sofosbuvir/daclatasvir could lower the risk of SARS-CoV-2 infection. Nitazoxanide was selected given its favourable pharmacokinetics and in vitro antiviral effects against SARS-CoV-2. Sofosbuvir/daclatasvir had shown favourable results in early clinical trials. In this clinical trial in Johannesburg, South Africa, healthcare workers and others at high risk of infection were randomized to 24 weeks of either nitazoxanide or sofosbuvir/daclatasvir as prevention, or standard prevention advice only. Participants were evaluated every 4 weeks for COVID-19 symptoms and had antibody and PCR testing. The primary endpoint was positive SARS-CoV-2 PCR and/or serology ≥7 days after randomization, regardless of symptoms. A Poisson regression model was used to estimate the incidence rate ratios of confirmed SARS-CoV-2 between each experimental arm and control. Between December 2020 and January 2022, 828 participants were enrolled. COVID-19 infections were confirmed in 100 participants on nitazoxanide (2234 per 1000 person-years; 95% CI 1837-2718), 87 on sofosbuvir/daclatasvir (2125 per 1000 person-years; 95% CI 1722-2622) and 111 in the control arm (1849 per 1000 person-years; 95% CI 1535-2227). There were no significant differences in the primary endpoint between the treatment arms, and the results met the criteria for futility. In the safety analysis, the frequency of grade 3 or 4 adverse events was low and similar across arms. In this randomized trial, nitazoxanide and sofosbuvir/daclatasvir had no significant preventative effect on infection with SARS-CoV-2 among healthcare workers and others at high risk of infection.


Territorial Strategy of Medical Units for Addressing the First Wave of the COVID-19 Pandemic in the Metropolitan Area of Mexico City: Analysis of Mobility, Accessibility and Marginalization.
The COVID-19 pandemic has caused an exponential increase in the demand for medical care worldwide. In Mexico, the COVID Medical Units (CMUs) conversion strategy was implemented. To evaluate the CMU coverage strategy in the Mexico City Metropolitan Area (MCMA) by territory. The CMU directory was used, as were COVID-19 infection and mobility statistics and Mexican 2020 census information at the urban geographic area scale. The degree of urban marginalization by geographic area was also considered. Using descriptive statistics and the calculation of a CMU accessibility index, population aggregates were counted based on coverage radii. In addition, two regression models are proposed to explain (1) the territorial and temporal trend of COVID-19 infections in the MCMA and (2) the mobility of the COVID-infected population visiting medical units. The findings of the evaluation of the CMU strategy were (1) in the MCMA, COVID-19 followed a pattern of contagion from the urban center to the periphery; (2) given the growth in the number of cases and the overload of medical units, the population traveled greater distances to seek medical care; (3) after the CMU strategy was evaluated at the territory level, it was found that 9 out of 10 inhabitants had a CMU located approximately 7 km away; and (4) at the metropolitan level, the lowest level of accessibility to the CMU was recorded for the population with the highest levels of marginalization, i.e., those residing in the urban periphery.


Specific properties of brown adipose tissue mitochondrial membrane.
1. Mitochondrial membrane of brown adipose tissue compared to that of liver possesses a very high activity of oxidative enzymes but a low activity of ATPase. 2. The polypeptide composition of the mitochondrial membranes proves that the above differences in enzyme activities are due to increased content of oxidative enzymes and decreased content of ATPase in brown adipose tissue. 3. The inhibition of ATPase of brown adipose tissue mitochondria by aurovertin, oligomycin and DCCD indicates modified proportions between the components of the ATPase complex. 4. The organization of brown adipose tissue mitochondrial membrane in relation to its thermogenic function is discussed.


Analysis of Insulin Resistance in Nonalcoholic Steatohepatitis.
Insulin resistance is a major phenotype observed in nonalcoholic steatohepatitis (NASH), the advanced stage of nonalcoholic fatty liver disease (NAFLD). Insulin resistance in NASH is characterized by reductions in whole body, hepatic, and adipose tissue insulin sensitivity. The mechanisms underlying hepatic insulin resistance is primarily associated with hepatic glucose production (HGP) rate. Hepatic insulin resistance can also be a consequence or a driving factor of hepatic lipid accumulation by increasing free fatty acid synthesis, delivery, and catabolism. The common method to assess hepatic insulin resistance is to measure hepatic glucose production (HGP) using isotope tracer distribution technique. However, non-radioactive approaches have been developed to assess hepatic insulin resistance in the context of NASH. In this chapter, we describe the methods to evaluate hepatic insulin resistance in animal models of NASH by examining insulin sensitivity and glucose tolerance as well as the key molecules in hepatic insulin signaling pathways.


Real-World Fatigue Testing in Professional Rugby Union: A Systematic Review and Meta-analysis.
Professional rugby union is a high-intensity contact sport with position-specific high training and match volumes across a season that may lead to periods of fatigue if above a typically experienced threshold. This study assesses the influence of match play and/or training on fatigue levels in rugby union players. We aimed to perform a systematic review and meta-analysis of measures used to assess fatigue status in male professional rugby union players. Using electronic databases (PubMed, SPORTDiscus, Web of Science, Cochrane Library, EMBASE, and MEDLINE), a systematic review of fatigue testing in rugby union was conducted on (1) neuromuscular, (2) subjective self-report, (3) biochemical, and (4) heart rate-derived measures. Thirty-seven articles were included in this systematic review, of which 14 were further included in a meta-analysis. The results of the meta-analysis revealed small, yet not significant, decreases in countermovement jump height immediately after (effect size [ES] = - 0.29; 95% confidence interval [CI] - 0.64 to 0.06), 24 h (ES = - 0.43; 95% CI - 3.99 to 3.21), and 48 h (ES = - 0.22; 95% CI - 0.47 to 0.02) after exposure to rugby union match play or training. Reported wellness (ES = - 0.33; 95% CI - 1.70 to 1.04) and tiredness (ES = - 0.14; 95% CI - 1.30 to 1.03) declined over a period of a few weeks (however, the results were not-statistically significant), meanwhile muscle soreness increased (ES = 0.91; 95% CI 0.06 to 1.75) within the 96 h after the exposure to rugby union match play or training. Finally, while cortisol levels (ES = 1.87; 95% CI - 1.54 to 5.29) increased, testosterone declined (ES = - 1.54; 95% CI - 7.16 to 4.08) within the 24 h after the exposure. However, these results were not statistically significant. Subjective measures of muscle soreness can be used to assess fatigue after match play and training in rugby union players. Within-study and between-study variability for countermovement jump height, biochemical markers, and heart rate-derived measures means the utility (practical application) of these measures to assess fatigue in professional rugby union players after matches and training is unclear. PROSPERO ID: CRD42020216706.


Postnatal expression of myosin isoforms in the genioglossus and diaphragm muscles.
We studied the expression of myosin heavy chain (MHC) and native myosin isoforms in the genioglossus (GG) and costal diaphragm (DIA) muscles of the rat during postnatal development using both denaturing and nondenaturing gel electrophoresis. Primary myotubes in both fast and slow muscles homogeneously express slow as well as embryonic myosin. Since the adult GG is comprised primarily of fast MHC isoforms, whereas the adult DIA is characterized by a mixture of MHC slow and fast isoforms, we hypothesized that the GG and DIA would be subject to different temporal patterns of MHC isoform expression during postnatal development. Native myosin and MHC gels demonstrated a persistence of neonatal MHC (MHC neo) on day 25 in the GG, whereas this isoform was not detected beyond day 21 in the DIA. The MHC phenotype in GG of the adult demonstrated a predominance of MHC 2X (35% +/- 8) and MHC 2B (45% +/- 10) with a smaller proportion of MHC 2A (19% +/- 5). In contrast, the MHC phenotype in adult DIA was characterized by approximately equal proportions of MHC slow (25% +/- 3), MHC 2A (34% +/- 10), and MHC 2X (31% +/- 12) with a small percentage of MHC 2B (9% +/- 7). These data suggest that postnatal regulation of MHC expression in the GG and DIA is muscle specific.


Pharmacological and anatomical evidence for an interaction between mGluR5- and GABA(A) alpha1-containing receptors in the discriminative stimulus effects of ethanol.
The discriminative stimulus properties of ethanol are mediated in part by positive modulation of GABA(A) receptors. Recent evidence indicates that metabotropic glutamate receptor subtype 5 (mGluR5) activity can influence GABA(A) receptor function. Therefore, the purpose of this work was to examine the potential involvement of mGluR5 in the discriminative stimulus effects of ethanol. In rats trained to discriminate ethanol (1 g/kg, intragastric gavage (i.g.)) from water, 2-methyl-6-(phenylethyl)-pyridine (MPEP) (1-50 mg/kg, i.p.) a selective noncompetitive antagonist of the mGlu5 receptor did not produce ethanol-like stimulus properties. However, pretreatment with MPEP (30 mg/kg) reduced the stimulus properties of ethanol as indicated by significant reductions in ethanol-appropriate responding, specifically at 0.5 and 1 g/kg ethanol, and a failure of ethanol test doses (1 and 2 g/kg) to fully substitute for the ethanol training dose. To test whether mGluR5 antagonism altered the GABA(A) receptor component of the ethanol stimulus, the ability of MPEP to modulate pentobarbital and diazepam substitution for ethanol was assessed. Pentobarbital substitution (1-10 mg/kg, i.p.) for ethanol was not altered by MPEP pretreatment. However, MPEP pretreatment inhibited the ethanol-like stimulus properties of diazepam (5 mg/kg, i.p.). To examine a potential anatomical basis for these pharmacological findings, expression patterns of mGluR5- and benzodiazepine-sensitive GABA(A) alpha1-containing receptors were examined by dual-label fluorescent immunohistochemistry with visualization by confocal microscopy. Results indicated that mGluR5- and GABA(A) alpha1-containing receptors were both coexpressed in limbic brain regions and colocalized on the same cells in specific brain regions including the amygdala, hippocampus, globus pallidus, and ventral pallidum. Together, these findings suggest an interaction between mGluR5- and benzodiazepine-sensitive GABA(A) receptors in mediating ethanol discrimination.


Simple and rapid detection of calcium peroxide in flour based on methanobactin peroxidase-like activity.
Calcium peroxide is forbidden to be added to flour as brightener in many countries. A rapid and sensitive spectrophotometric method for the detection of calcium peroxide in flour was proposed. Methanobactin (Mb), a copper-binding small peptide of methanotrophs with excellent peroxidase-like activity, has been successfully applied for H2O2-mediated co-oxidation between phenol and 4-aminoantipyrine to give a colored product that can be detected at 505 nm. When the concentration of Mb-Cu was 6.5 × 10-6 mol/L, the detection temperature was 50 ℃, and the detection time was 5 min, the linear range for quantification of calcium peroxide concentration was observed between 0.4 and 10.0 mg/L with R2 = 0.99397. The limit of detection was 0.027 mg/L (3.34 mg/kg flour) and the average recovery of standard addition was 99.6%-100.5%. Mb was very stable and still maintained catalytic activity even at 50 ℃ in acidic medium, which gave the proposed method has simple process and rapid detection speed.


Remodeling of the ARID1A tumor suppressor.
In recent years, AT-rich interactive domain-containing protein 1A (ARID1A) has been widely accepted as a bona fide tumor suppressor due to its essential role in preventing tumorigenesis and tumor progression in both mouse and human contexts. ARID1A shows high mutation frequencies in both cancers and preneoplastic lesions. The loss of ARID1A expression in cancer cells leads to increases in cell proliferation, invasion and migration and reductions in cell apoptosis and chemosensitivity. The tumor-suppressive role of ARID1A is mainly attributed to its regulation of gene transcription, which can be induced either directly by chromatin remodeling or indirectly by affecting histone modifications. ARID1A also acts independently of its cardinal transcription-regulating mechanisms, which include interfering with protein-protein interactions. Interestingly, nonmutational mechanisms, such as regulation by DNA hypermethylation, microRNAs, and ubiquitinases/deubiquitinases, have provided another perspective on ARID1A inactivation in cancer. Since the critical tumor-suppressive role of ARID1A has been revealed, several studies have attempted to identify synthetic lethal targets with ARID1A mutation/inactivation as an alternative strategy for cancer treatment.


Estimating individual treatment effects on COPD exacerbations by causal machine learning on randomised controlled trials.
Estimating the causal effect of an intervention at individual level, also called individual treatment effect (ITE), may help in identifying response prior to the intervention. We aimed to develop machine learning (ML) models which estimate ITE of an intervention using data from randomised controlled trials and illustrate this approach with prediction of ITE on annual chronic obstructive pulmonary disease (COPD) exacerbation rates. We used data from 8151 patients with COPD of the Study to Understand Mortality and MorbidITy in COPD (SUMMIT) trial (NCT01313676) to address the ITE of fluticasone furoate/vilanterol (FF/VI) versus control (placebo) on exacerbation rate and developed a novel metric, Q-score, for assessing the power of causal inference models. We then validated the methodology on 5990 subjects from the InforMing the PAthway of COPD Treatment (IMPACT) trial (NCT02164513) to estimate the ITE of FF/umeclidinium/VI (FF/UMEC/VI) versus UMEC/VI on exacerbation rate. We used Causal Forest as causal inference model. In SUMMIT, Causal Forest was optimised on the training set (n=5705) and tested on 2446 subjects (Q-score 0.61). In IMPACT, Causal Forest was optimised on 4193 subjects in the training set and tested on 1797 individuals (Q-score 0.21). In both trials, the quantiles of patients with the strongest ITE consistently demonstrated the largest reductions in observed exacerbations rates (0.54 and 0.53, p<0.001). Poor lung function and blood eosinophils, respectively, were the strongest predictors of ITE. This study shows that ML models for causal inference can be used to identify individual response to different COPD treatments and highlight treatment traits. Such models could become clinically useful tools for individual treatment decisions in COPD.


Risk factors of presenile nuclear cataract in health screening study.
To identify risk factors for the development of presenile nuclear cataract in health screening test. The cross sectional study included a total of 532 eyes of 266 participants aged 30 to 49 years of Samsung Medical Center from February 2013 to April 2015. Presence of nuclear cataract was defined when the log MAR visual acuity with correction was greater than or equal to 0.2 and one or more of the following were met: Pentacam Nuclear Staging (PNS) grading score ≥ 1, average value of nuclear density ≥ 15%, maximum value of nuclear density ≥ 30%. Possible risk factors were obtained from blood tests and questionnaires of a health screening test of Samsung Medical Center. Association between nuclear cataract and risk factors was investigated using univariate and multivariate logistic regression analysis by generalized estimating equation (GEE) models. Five factors were significantly associated with presenile nuclear cataract: current smoking [odds ratio (OR) = 2.80, 95% confidence interval (CI), 1.10-7.12, p = 0.0310], non-exercise and high amount of daily physical exercise (OR = 3.99, 95% CI, 1.27-12.52, p = 0.0178; OR = 2.92, 95% CI, 1.38-6.22, p = 0.0053), asthma (OR = 8.93, 95% CI, 1.12-71.15, p = 0.0386), tuberculosis (OR = 4.28, 95% CI, 1.36-13.50, p = 0.0131), and higher total iron binding capacity (OR = 1.01, 95% CI, 1.00-1.02, p = 0.0059). Presenile nuclear cataract is related to current smoking, non-exercise or high amount of physical exercise, asthma, tuberculosis, and iron deficiency status. The association of non-exercise group and presenile nuclear cataract seems to be related to co-morbidity. Patients with asthma, tuberculosis, or iron deficiency anemia are recommended to receive frequent ophthalmic examination to detect cataract.


Progastrin: a potential predictive marker of liver metastasis in colorectal cancer.
Staging of colorectal cancer often fails to discriminate outcomes of patients with morphologically similar tumours that exhibit different clinical behaviours. Data from several studies suggest that the gastrin family of growth factors potentiates colorectal cancer tumourigenesis. The aim of this study was to investigate whether progastrin expression may predict clinical outcome in colorectal cancer. Patients with colorectal adenocarcinoma of identical depth of invasion who had not received neoadjuvant therapy were included. The patients either had stage IIa disease with greater than 3-year disease-free survival without adjuvant therapy or stage IV disease with liver metastases on staging CT. Progastrin expression in tumour sections was scored with reference to the intensity and area of immunohistochemical staining. Progastrin expression by stage IV tumours was significantly greater than stage IIa tumours with mean progastrin immunopositivity scores of 2.1 ± 0.2 versus 0.5 ± 0.2, respectively (P < 0.001). This is the first study to show that progastrin expression may be predictive of aggressive tumour behaviour in patients with colorectal cancer and supports its clinical relevance and potential use as a biomarker.


The role of income inequality on consumption-based greenhouse gas emissions under different stages of economic development.
In recent decades, a large number of authors investigated whether income inequality level and climate change were compatible. However, the relationship between inequality level impact and consumption-based greenhouse gas (GHG) emissions was scarcely analysed. Thus, the objective of the present paper was to analyse the relationship between income inequality level and consumption-based GHG emissions from 1990 to 2014 under different stages of economic development. The results revealed a large gap between consumption-based GHG emissions at different economic development stages. Economic growth contributed to the growth of consumption-based GHG emissions in all income groups, and the largest effect was observed in middle-low- and low-income countries. Higher income inequality level was associated with lower negatively influenced consumption-based emissions in high- and middle-high-income countries, confirming the marginal propensity to emit and 'Veblen effect' theories. Meanwhile, regarding the middle-low- and low-income groups, the relationships between inequality level and GHG emissions were insignificant. Urbanisation level contributed to the growth of consumption-based GHG emissions in middle- and low-income countries. Education level contributed to the reduction of consumption-based GHG emissions in high-, middle-high-, and middle-low-income countries, and growth of emissions in low-income countries. Therefore, our analysis revealed that income inequality and climate change are still incompatible in all income groups, and we suggested how to achieve the reduction of income inequality and climate change simultaneously.


Integrin Antibody Decreases Deformability of Patient-Derived Pre-B Acute Lymphocytic Leukemia Cells as Measured by High-Frequency Acoustic Tweezers.
This article reports a study of cell mechanics in patient-derived (primary) B-cell acute lymphocytic leukemia (ALL) cells treated with antibodies against integrins. Leukemia cell adhesion to stromal cells mediates chemotherapeutic drug resistance, also known as cell adhesion-mediated chemotherapeutic drug resistance. We have previously shown that antibodies against integrin α4 and α6 adhesion molecules can de-adhere ALL cells from stromal cells or counter-receptors. Because drug-resistant cells are more deformable, as evaluated by single-beam acoustic tweezers, we hypothesized that changes in cell mechanics might contribute to the de-adhesive effect of integrin-targeting antibodies. In this study, the deformability of primary pre-B ALL cells was evaluated by single-beam acoustic tweezers after treatments with the de-adhering antibody Tysabri or P5G10 against integrin α4 and α6 adhesion molecules. We demonstrated that primary ALL cells treated with P5G10 expressed decreased deformability compared with immunoglobulin G1 -treated control cells (P < .05). Tysabri did not show an effect on deformability (P > .05). These results suggest that decreased deformability is associated with an integrin α6 blockade. Further assessments of the functional roles of deformability and integrin blockades in B-ALL cell drug resistance and deformability, respectively, are necessary.


Comparative assessment of 6-[18 F]fluoro-L-m-tyrosine and 6-[18 F]fluoro-L-dopa to evaluate dopaminergic presynaptic integrity in a Parkinson's disease rat model.
Because of the progressive loss of nigro-striatal dopaminergic terminals in Parkinson's disease (PD), in vivo quantitative imaging of dopamine (DA) containing neurons in animal models of PD is of critical importance in the preclinical evaluation of highly awaited disease-modifying therapies. Among existing methods, the high sensitivity of positron emission tomography (PET) is attractive to achieve that goal. The aim of this study was to perform a quantitative comparison of brain images obtained in 6-hydroxydopamine (6-OHDA) lesioned rats using two dopaminergic PET radiotracers, namely [18 F]fluoro-3,4-dihydroxyphenyl-L-alanine ([18 F]FDOPA) and 6-[18 F]fluoro-L-m-tyrosine ([18 F]FMT). Because the imaging signal is theoretically less contaminated by metabolites, we hypothesized that the latter would show stronger relationship with behavioural and post-mortem measures of striatal dopaminergic deficiency. We used a within-subject design to measure striatal [18 F]FMT and [18 F]FDOPA uptake in eight partially lesioned, eight fully lesioned and ten sham-treated rats. Animals were pretreated with an L-aromatic amino acid decarboxylase inhibitor. A catechol-O-methyl transferase inhibitor was also given before [18 F]FDOPA PET. Quantitative estimates of striatal uptake were computed using conventional graphical Patlak method. Striatal dopaminergic deficiencies were measured with apomorphine-induced rotations and post-mortem striatal DA content. We observed a strong relationship between [18 F]FMT and [18 F]FDOPA estimates of decreased uptake in the denervated striatum using the tissue-derived uptake rate constant Kc . However, only [18 F]FMT Kc succeeded to discriminate between the partial and the full 6-OHDA lesion and correlated well with the post-mortem striatal DA content. This study indicates that the [18 F]FMT could be more sensitive, with respect of [18 F]FDOPA, to investigate DA terminals loss in 6-OHDA rats, and open the way to in vivo L-aromatic amino acid decarboxylase activity targeting in future investigations on progressive PD models.


Elbow Release and Tricepsplasty in Arthrogrypotic Patients: A Long-Term Follow-Up Study.
Obtaining elbow flexion to improve hand-to-mouth reach capability is an essential component of achieving functional independence in pediatric patients with arthrogryposis. This study analyzed the long-term outcomes of elbow release and tricepsplasty in a series of children with arthrogryposis at a tertiary institution. Medical records of patients with arthrogryposis who underwent elbow release and tricepsplasty from 1993 to 2015, with at least 2 years of follow-up, were reviewed. Collected measures included preoperative elbow passive range of motion (ROM), postoperative elbow passive and active ROM, shoulder passive and active ROM, and Pediatric Outcomes Data Collection Instrument (PODCI) scores. Our analysis compared pre- and postoperative follow-up of elbow passive ROM and reviewed PODCI scores with age-adjusted normative values. Seventeen patients (4 female and 13 male) with 24 affected upper extremities (10 left elbow and 14 right elbow) were included in final analysis. Age at final follow-up averaged 11 years (range, 4-20 years), mean age at surgery was 2.7 years (range, 9.6 months-9.3 years) with mean follow-up by extremity at 8.3 years (range, 2-18 years). Differences in pre- and post-operative passive elbow ROM were significant for extension, flexion, and total arc of motion. Most parent and self-reported PODCI scores were less than the age-adjusted normal population, except in the domains of Comfort and Pain and Happiness. Long-term follow-up of elbow release and tricepsplasty in patients with arthrogryposis indicates both increased and sustained elbow flexion and arc of motion. Although PODCI scores were lower compared with the age-adjusted norm, pediatric patients with arthrogryposis were just as happy and had no more discomfort than their unaffected age-adjusted norms. Therapeutic IV.


A Thylakoid Membrane Protein Functions Synergistically with GUN5 in Chlorophyll Biosynthesis.
Chlorophyll (Chl) is essential for photosynthetic reactions and chloroplast development. While the enzymatic pathway for Chl biosynthesis is well established, the regulatory mechanism underlying the homeostasis of Chl levels remains largely unknown. In this study, we identified CBD1 (Chlorophyll Biosynthetic Defect1), which functions in the regulation of chlorophyll biosynthesis. The CBD1 gene was expressed specifically in green tissues and its protein product was embedded in the thylakoid membrane. Furthermore, CBD1 was precisely co-expressed and functionally correlated with GUN5 (Genome Uncoupled 5). Analysis of chlorophyll metabolic intermediates indicated that cbd1 and cbd1gun5 mutants over-accumulated magnesium protoporphyrin IX (Mg-Proto IX). In addition, the cbd1 mutant thylakoid contained less Mg than the wild type not only as a result of lower Chl content, but also implicating CBD1 in Mg transport. This was supported by the finding that CBD1 complemented a Mg2+ uptake-deficient Salmonella strain under low Mg conditions. Taken together, these results indicate that CBD1 functions synergistically with CHLH/GUN5 in Mg-Proto IX processing, and may serve as a Mg-transport protein to maintain Mg homeostasis in the chloroplast.


Changes in calcium fluxes in mitochondria, microsomes, and plasma membrane vesicles of livers from monosodium L-glutamate-obese rats.
The purpose of this work was to evaluate if the fat liver accumulation interferes with intracellular calcium fluxes and the liver glycogenolytic response to a calcium-mobilizing α(1)-adrenergic agonist, phenylephrine. The animal model of monosodium L-glutamate (MSG)-induced obesity was used. The adult rats develop obesity and steatosis. Calcium fluxes were evaluated through measuring the (45)Ca(2+) uptake by liver microsomes, inside-out plasma membrane, and mitochondria. In the liver, assessments were performed on the calcium-dependent glycogenolytic response to phenylephrine and the glycogen contents. The Ca(2+) uptake by microsomes and plasma membrane vesicles was reduced in livers from obese rats as a result of reduction in the Ca(2+)-ATPase activities. In addition, the plasma membrane Na(+)/K(+)-ATPase was reduced. All these matched effects could contribute to elevated resting intracellular calcium levels in the hepatocytes. Livers from obese rats, albeit smaller and with similar glycogen contents to those of control rats, released higher amounts of glucose in response to phenylephrine infusion, which corroborates these observations. Mitochondria from obese rats exhibited a higher capacity of retaining calcium, a phenomenon that could be attributed to a minor susceptibility of the mitochondrial permeability transition pore opening.


Improved library preparation with the new iCLIP2 protocol.
Individual-nucleotide resolution UV crosslinking and immunoprecipitation (iCLIP) is a state-of-the-art technology to map the RNA interaction sites of an RNA-binding protein (RBP) across the transcriptome. Here, we present the new iCLIP2 protocol that allows to obtain high-quality iCLIP libraries in a fast and efficient manner. The new protocol comprises separate adapter ligations, two cDNA amplification steps and bead-based size selection. The full procedure can be completed within four days. Our advances significantly increase the complexity of the iCLIP2 libraries, resulting in a more comprehensive representation of RBP binding sites. Overall, the methodological advances in iCLIP2 allow efficient library generation and thereby promote the versatile and flexible application of this important technology.


Apoptosis signal-regulating kinase 1 is an intracellular inducer of p38 MAPK-mediated myogenic signalling in cardiac myoblasts.
Myogenic differentiation is an essential process for the myogenesis in response to various extracellular stimuli. p38 MAPK is a core signalling molecule in myogenic differentiation. The activation of p38 MAPK is required for myogenic differentiation; however, the mechanism for this activation remains undefined. ASK1 is a member of the MAP3K family that activates both JNK and p38 MAPK pathways in response to an array of stresses such as oxidative stress, endoplasmic reticulum stress and calcium influx. Here, we reported that TNFα was significantly released from H9c2 cardiac myoblast in differentiation medium. Furthermore, the oxidant H(2)O(2) acted as a messenger in the TNFα signalling pathway to disrupt the complex of ASK1-Trx, which was followed by the activation of ASK1 in cardiac myogenic differentiation. Subsequently, the activated ASK1 stimulated MKK3/6-p38MAPK signalling cascade to induce specific myogenic differentiation. In addition, exogenous TNFα added to the medium at physiological levels enhanced the ASK1-p38 MAPK signalling pathway through the increased generation of H(2)O(2). Interestingly, inhibition of p38 MAPK abrogated the production of H(2)O(2), suggesting that there might be a positive feedback loop in the myogenic-redox signalling pathway. These results indicate that ASK1 is a new intracellular regulator of activation of the p38 MAPK in cardiac myogenic differentiation.


The stiffness of the flagella of impaled bull sperm.
The elastic rigidity (stiffness) of impaled motionless bull sperm flagella has been determined by a manipulatory technique which permitted direct analytical treatment of the experimental system. The effects of external ATP and ADP were measured. It was found that ATP acts as a plasticizing agent, while ADP does not. The stiffness measured for flagella in a medium without ATP was 15 times greater than the value measured with 10 mM ATP present. The rigor-like stiffness measured with no ATP present is reversible with ATP and seems to be correlated to a transition in the state of the contractile system.


A knockout cell library of GPI biosynthetic genes for functional studies of GPI-anchored proteins.
Over 100 kinds of proteins are expressed as glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) on the cell surface in mammalian cells. GPI-APs possess unique properties in terms of their intracellular trafficking and association with lipid rafts. Although it is clear that GPI-APs play critical roles in various biological phenomena, it is poorly understood how the GPI moiety contributes to these mechanisms. More than 30 genes are involved in the correct biosynthesis of GPI-APs. We here constructed a cell library in which 32 genes involved in GPI biosynthesis were knocked out in human embryonic kidney 293 cells. Using the cell library, the surface expression and sensitivity to phosphatidylinositol-specific phospholipase C of GPI-APs were analyzed. Furthermore, we identified structural motifs of GPIs that are recognized by a GPI-binding toxin, aerolysin. The cell-based GPI-knockout library could be applied not only to basic researches, but also to applications and methodologies related to GPI-APs.


Congenital medulloblastoma: an inquiry into origins.
An infant aged 8 h died from cardiorespiratory causes. At autopsy, a microscopic midline cerebellar medulloblastoma was found. Neoplastic cells occurred within the fetal external granular cell layers, with invasion of perivascular spaces as well as the molecular and internal granular cell layers. This extremely rare occurrence lends support to the hypothesis that cerebellar medulloblastoma occurring in older patients originates within retained rests of primitive cells in the fetal external granular cell layer.


Synthesis of building blocks for solid-phase introduction of diethylenetriaminepentaacetic acid (DTPA) to oligonucleotides and oligopeptides.
Synthesis of building blocks that allow site-specific incorporation of diethylenetriaminepentaacetic acid (DTPA) to oligonucleotides and oligopeptides using phosphoramidite and Fmoc chemistries, respectively, is described.


Gold nanoclusters: An ultrasmall platform for multifaceted applications.
Gold nanoclusters (Au NCs) with a core size below 2 nm form an exciting class of functional nano-materials with characteristic physical and chemical properties. The properties of Au NCs are more prominent and extremely different from their bulk counterparts. The synthesis of Au NCs is generally assisted by template or ligand, which impart excellent cluster stability and high quantum yield. The tunable and sensitive physicochemical properties of Au NCs open horizons for their advanced applications in various interdisciplinary fields. In this review, we briefly summarize the solution phase synthesis and origin of the characteristic properties of Au NCs. A vast review of recent research work introducing biosensors based on Au NCs has been presented along with their specifications and detection limits. This review also highlights recent progress in the use of Au NCs as bio-imaging probe, enzyme mimic, temperature sensing probe and catalysts. A speculation on present challenges and certain future prospects have also been provided to enlighten the path for advancement of multifaceted applications of Au NCs.