(H)Elping nerve growth factor: Elp1 inhibits TrkA's phosphatase to maintain retrograde signaling.
Nerve growth factor (NGF) regulates many aspects of neuronal biology by retrogradely propagating signals along axons to the targets of those axons. How this occurs when axons contain a plethora of proteins that can silence those signals has long perplexed the neurotrophin field. In this issue of the JCI, Li et al. suggest an answer to this vexing problem, while exploring why the Elp1 gene that is mutated in familial dysautonomia (FD) causes peripheral neuropathy. They describe a distinctive function of Elp1 as a protein that is required to sustain NGF signaling by blocking the activity of its phosphatase that shuts off those signals. This finding helps explain the innervation deficits prominent in FD and reveals a unique role for Elp1 in the regulation of NGF-dependent TrkA activity.


Stable and wavelength-tunable silicon-micro-ring-resonator based erbium-doped fiber laser.
In this work, we propose and demonstrate a stable and wavelength-tunable erbium-doped fiber (EDF) ring laser. Here, a silicon-on-insulator (SOI)-based silicon-micro-ring-resonator (SMRR) is used as the wavelength selective element inside the fiber ring cavity. A uniform period grating coupler (GC) is used to couple between the SMRR and single mode fiber (SMF) and serves also as a polarization dependent element in the cavity. The output lasing wavelength of the proposed fiber laser can be tuned at a tuning step of 2 nm (defined by the free spectral range (FSR) of the SMRR) in a bandwidth of 35.2 nm (1532.00 to 1567.20 nm), which is defined by the gain of the EDF. The optical-signal-to-noise-ratio (OSNR) of each lasing wavelength is larger than 42.0 dB. In addition, the output stabilities of power and wavelength are also discussed.


Dynamics of Anguillicola crassus (Nematoda: Dracunculoidea) larval infection in paratenic host fishes of Lake Balaton, Hungary.
Anguillicola crassus, a nematode parasitizing the swimbladder of the European eel (Anguilla anguilla) caused substantial mortality among eels in Lake Balaton in the years 1991 and 1992. Parallel to a 3-year study of the infection dynamics of eels, Anguillicola larval infection of paratenic host fishes was also surveyed in the lake between 1991 and 1993. During that study, a total of 1,382 specimens of 22 paratenic host fish species were processed. The results showed that anguillicolosis had become a parasitosis widespread throughout the lake, and larval infection could be detected in practically all paratenic hosts examined. The prevalence and intensity values recorded in the paratenic hosts do not completely follow the dynamic change observed during the survey of eel infection. During the study of larval infection in paratenic hosts, only inter-species differences in the prevalence and intensity of infection could be found, rather than the expected course of infection spreading from the West to the East in both space and time. Marked differences existed between paratenic host species in the degree of host reaction to the larvae.


Deletion of C2orf34, PREPL and SLC3A1 causes atypical hypotonia-cystinuria syndrome.
Hypotonia-cystinuria syndrome (HCS) and 2p21 deletion syndrome are two recessive contiguous gene deletion syndromes associated with cystinuria type I. The deletions differ in size and the number of genes involved. In HCS patients, only SLC3A1 and PREPL are disrupted. In the 2p21 deletion syndrome, two additional genes (C2orf34 and PPM1B) are lost. Clinical and molecular analysis of two siblings who presented with an atypical HCS phenotype. Molecular analysis of the SLC3A1/PREPL locus was performed in the patients using quantitative polymerase chain reaction (PCR) methods. HCS in both siblings was confirmed with the deletion screen of the SLC3A1/PREPL locus. Fine mapping of the breakpoint revealed a deletion of 77.4 kb, including three genes: SLC3A1, PREPL and C2orf34. Features not present in classical HCS were a mild/moderate mental retardation and a respiratory chain complex IV deficiency documented in patient 2. We report the first patients with a deletion of SLC3A1, PREPL and C2orf34. They present with a phenotype intermediate between HCS and 2p21 deletion syndrome. These patients facilitate the elucidation of the contribution of each gene to the phenotype in the different 2p21 deletion syndromes.


Cephalometric studies of the mandible, its masticatory muscles and vasculature of growing Göttingen Minipigs-A comparative anatomical study to refine experimental mandibular surgery.
Over many decades, the Göttingen Minipig has been used as a large animal model in experimental surgical research of the mandible. Recently several authors have raised concerns over the use of the Göttingen Minipig in this research area, observing problems with post-operative wound healing and loosening implants. To reduce these complications during and after surgery and to improve animal welfare in mandibular surgery research, the present study elucidated how comparable the mandible of minipigs is to that of humans and whether these complications could be caused by specific anatomical characteristics of the minipigs' mandible, its masticatory muscles and associated vasculature. Twenty-two mandibular cephalometric parameters were measured on CT scans of Göttingen Minipigs aged between 12 and 21 months. Ultimately, we compared this data with human data reported in the scientific literature. In addition, image segmentation was used to determine the masticatory muscle morphology and the configuration of the mandibular blood vessels. Compared to data of humans, significant differences in the mandibular anatomy of minipigs were found. Of the 22 parameters measured only four were found to be highly comparable, whilst the others were not. The 3D examinations of the minipigs vasculature showed a very prominent deep facial vein directly medial to the mandibular ramus and potentially interfering with the sectional plane of mandibular distraction osteogenesis. Damage to this vessel could result in inaccessible bleeding. The findings of this study suggest that Göttingen Minipigs are not ideal animal models for experimental mandibular surgery research. Nevertheless if these minipigs are used the authors recommend that radiographic techniques, such as computed tomography, be used in the specific planning procedures for the mandibular surgical experiments. In addition, it is advisable to choose suitable age groups and customize implants based on the mandibular dimensions reported in this study.


Geographical distribution of family physicians in Japan: a nationwide cross-sectional study.
Geographical maldistribution of physicians, and their subsequent shortage in rural areas, has been a serious problem in Japan and in other countries. Family Medicine, a new board-certified specialty started 10 years ago in Japan by Japan Primary Care Association (JPCA), may be a solution to this problem. We obtained the workplace information of 527 (78.4%) of the 672 JPCA-certified family physicians from an online database. From the national census data, we also obtained the workplace information of board-certified general internists, surgeons, obstetricians/gynaecologists and paediatricians and of all physicians as the same-generation comparison group (ages 30 to 49). Chi-squared test and residual analysis were conducted to compare the distribution between family physicians and other specialists. Five hundred nineteen JPCA-certified family physicians and 137,587 same-generation physicians were analysed. The distribution of family physicians was skewed to municipalities with a lower population density, which shows a sharp contrast to the urban-biased distribution of other specialists. The proportion of family physicians in non-metropolitan municipalities was significantly higher than that expected based on the distribution of all same-generation physicians (p < 0.001). Family physicians distributed in favour of rural areas much more than any other specialists in Japan. The better balance of family physician distribution reported from countries with a strong primary care orientation seems to hold even in a country where primary care orientation is weak, physician distribution is not regulated, and patients have free access to healthcare. Family physicians comprise only 0.2% of all Japanese physicians. However, if their population grows, they can potentially rectify the imbalance of physician distribution. Government support is mandatory to promote family medicine in Japan.


Genetic variants in three genes and smoking show strong associations with susceptibility to exudative age-related macular degeneration in a Chinese population.
The present study was undertaken to replicate the associations of representative polymorphisms in three genes (complement factor H (CFH), complement factor B (BF) and HtrA serine peptidase 1 (HTRA1)) with exudative age-related macular degeneration (AMD) in a Han Chinese population, and to test if the modifiable environmental factors affect AMD susceptibility associated with different type of genotype in these genes. An age, gender and ethnicity matched case-control study was conducted to genotype the representative single neucleotide polymorphisms (SNPs) loci including rs1061170 and rs1410996 in CFH, rs641153 and rs4151667 in BF and rs11200638 in HTRA1 gene in 144 exudative AMD patients and 126 normal controls using PCR-RFLP and direct resequencing. The demographic characteristics and behavioral risk factors were also recorded. Allelic and genotypic associations for individual SNP and joint associations with two loci were performed. The gene-gene and gene-environment interactions were analyzed using multivariate non-conditional Logistic regression analysis. The C risk allele frequencies for CFH Y402H (rs1061170) in cases and controls were 12.5% and 5.4% respectively, which were much lower than those in Caucasians (P < 0.001). Compared with TT homozygous genotype, the CT heterozygous genotype was positively associated with AMD with odds ratio (OR) of 3.23 (1.36 - 5.07). However, the population attributable risk (PAR) of C allele was only 3.3% (1.4% - 4.3%). rs1410996 was also associated with AMD independent of Y402H. The ORs of exudative AMD for individuals carrying one copy risk allele and two copy risk alleles were 2.57 (1.21 - 5.45) and 4.76 (2.15 - 10.55) respectively, with correspondent PARs of 28.3% (2.0% - 40.5%) and 38.2% (21.8% - 45.4%). rs11200638 in HTRA1 was another susceptible locus for AMD and the risk homozygotes were significantly susceptible for exudutive AMD (OR = 3.98, 1.88 - 8.43) with PAR of 38.9% (24.3% - 45.8%). Education status and cigarette smoking were also related to exudative AMD. After controlling for environmental risk factors, CFH and HTRA1 SNPs were independently associated with exudative AMD, with OR of 3.50 (1.45 - 8.45) for CT genotype in Y402H, 3.34 (1.33 - 8.36) for GG genotype in rs1410996 and 3.85 (1.58 - 9.42) for AA genotype in rs11200638 respectively. The interaction analysis between gene and environmental factors showed that smoking synergistically increased susceptibility of AMD for heterozygotes of rs1410996, with OR(interaction) of 7.33 (P(interaction) = 0.029). In a Han Chinese population, CFH and HTRA1 polymorphisms appear to be independently and possibly additively hereditary contributors to exudative AMD. Y402H polymorphism conferred a significant but relatively lower contribution in Chinese than in Caucasians with a low frequency of risk allele. The gene-environment interaction may be a best way to encourage those with a high genetic risk to prevent AMD by avoiding modifiable factors until there is effective treatment for AMD.


Bosons outperform fermions: The thermodynamic advantage of symmetry.
We examine a quantum Otto engine with a harmonic working medium consisting of two particles to explore the use of wave function symmetry as an accessible resource. It is shown that the bosonic system displays enhanced performance when compared to two independent single particle engines, while the fermionic system displays reduced performance. To this end, we explore the trade-off between efficiency and power output and the parameter regimes under which the system functions as engine, refrigerator, or heater. Remarkably, the bosonic system operates under a wider parameter space both when operating as an engine and as a refrigerator.


Reduction of radiation dose in pediatric brain CT is not associated with missed injuries or delayed diagnosis.
Increased accuracy of CTs in the identification of traumatic injuries compared with physical examination or conventional radiography is well documented. Our goal was to identify the most effective strategy for decreasing radiation exposure while retaining the benefits of computerized imaging. Based on a literature review and our trauma registry, the mortality risk of untreated injuries was compared with that of patients who received treatment of injuries diagnosed by CT. Because automated exposure control of tube current is not routinely used with brain CT, this region was identified as the initial focus for a dose-saving algorithm. CT settings were adjusted for children studies and the new settings were implemented into four protocols based on age. Images were compared and reviewed by radiologists for the ability to identify traumatic injuries. Effective dose (ED) was estimated using Monte Carlo simulations. The lifetime incidence and mortality for thyroid cancer and leukemia were assessed. In-hospital mortality of unidentified injury in trauma patients is 8.0%. Forty dose-saving CTs were performed and no injuries were missed. The ED decreased by 5.2-, 4.5-, 2.62-, and 2.5-fold in each group. Decreasing the ED is achievable, theoretically decreases the cancer risk and does not increase the missed injury rate.


Evaluation of Opening Offers Early for Deceased Donor Kidneys at Risk of Nonutilization.
Reducing non-utilization of kidneys recovered from deceased donors is a current policy concern for kidney allocation in the United States. The likelihood of non-utilization is greater with a higher kidney donor risk index (KDRI) offer. We examine how opening offers for organs with KDRI>1.75 to the broader waitlist at varying points of time affects usage rates. We simulate kidney allocation using data for 2018/01-2019/06 from Organ Procurement and Transplantation Network. For the simulation experiment, allocation policy is modified so that KDRI>1.75 organs are offered to all local candidates (same donation service area) after a set amount of cold time simultaneously. Open offers to candidates nationally are similarly examined. Simulation results (n=50 replications) estimate that opening offers locally for KDRI > 1.75 after 10 hours yields a non-utilization rate of 38% (range: 35%-42%), less than the prevailing rate of 55% of KDRI > 1.75 kidneys. Opening offers after 5 hours yields 30% (range: 26%-34%), reducing the prevailing non-utilization rate by 45%. Opening offers nationally after 10 and 5 hours yields non-utilization rates of 11% (range: 8%-15%) and 6% (range: 4%-9%) for KDRI>1.75 kidneys, respectively. Simulation findings indicate that opening offers and adjusting their timing can significantly reduce non-utilization of high-KDRI kidneys.


Characterization of the novel temperate Escherichia coli phage phiStx2k.
A novel temperate phage, phiStx2k, was induced from a clinical Escherichia coli isolate producing Shiga toxin (Stx) 2k. The phage particles have an icosahedral head (50 nm in diameter) and a long non-contractile tail (149 nm long). The phage genome consists of 46,647 bp of double-stranded DNA with an average G + C content of 51%. Genome sequence comparisons suggested that phiStx2k represents a new genus in the class Caudoviricetes. phiStx2k was capable of converting non-Stx-producing E. coli strains to Stx producers. These results expand our knowledge on the characteristics of Stx phages and highlight the potential risks of the emergence of Stx-producing strains or novel pathogens via horizontal gene transfer.


Neuroendocrine gastro-enteropancreatic tumors - from eminence based to evidence-based medicine - A Scandinavian view.
Neuroendocrine tumors (NETs) comprise a heterogenous group of neoplasms with variable clinical expression and progression. The primary tumors most frequently occur in the lungs, intestine and the pancreas. The NET incidence is approximately 6.1/100,000 per year with a prevalence higher than 35/100,000 per year. A NET may be functioning with symptoms related to hormone overproduction or non-functioning, not presenting any hormone-related symptoms. From the early 1980s and onwards, Uppsala University Hospital has contributed significantly to diagnosis, just to mention immunohistochemistry, radio-immunoassays for hormones and peptides and molecular imaging. On the therapeutic side, treatments with cytotoxics as well as biologicals such as, somatostatin analogs and interferons have been evaluated. We have furthermore been involved in important phase III trials for registration of so called, new targeted agents such as, RADIANT-3 and RADIANT-2. Our group were also the first to localize the gene for MEN I on chromosome 11 locus q13. Most recent developments have been the establishments of new biomarkers such as, olfactory receptor E51E1 as well as micro-RNAs in carcinoid tumors of the intestine and lung. A new oncolytic virus, Ad-Vince, for treatment of most NETs has been developed and is ready for the clinic. Furthermore, we have been involved in establishing Nordic and international collaborations. Today, NETs is an area with rapid development and recognized by international organizations at conferences, with large attendance. The Nordic countries continue to be significant contributors to the field.


Dative sigma- and pi-bonding in boron-nitrogen compounds: molecular structures of (CH3)2NB(CH3)N(CH3)B(CH3)2 and [(CH3)2B]2NN(CH3)2 determined by gas electron diffraction and quantum chemical calculations.
The molecular structures of dimethylamino[(dimethylboryl)methylamino]methylborane, Me2NBMeNMeBMe2 (1) and 1,1-bis(dimethylboryl)-2,2-dimethylhydrazine, (Me2B)2NNMe2 (2) have been determined by gas electron diffraction (GED), density functional theory calculations at the B3PW91/6-311++G** level and ab initio calculations at the MP2/6-311++G** level. 1 adopts an open structure similar to that of the isoelectronic hydrocarbon molecule permethylbutadiene; the central B-N bond distance at 148.0/149.3(7) pm (MP2/GED) corresponds to a single covalent N--B bond distance, the two terminal distances, 140.9/140.5(4) pm and 141.8/141.3(4) pm, correspond to the distance between N and B atoms joined by a covalent sigma-bond and a dative pi-bond. A closed form where the establishment of a dative bond between the terminal N and B atoms has led to the formation of a four-membered ring also corresponds to a minimum on the potential energy surface, but the energy is calculated to be 14.3 kJ mol(-1) higher at the MP2 level. This structure is also incompatible with the GED data. 2 adopts a structure in which a dative sigma-bond between the dimethylamino N atom and one of the boron atoms has led to the formation of a three-membered N(2)B ring. The dative sigma-bond distance is 165.5/164.0(13) pm, the two other bond distances in the ring are N-B=150.6/148.9(9) pm corresponding to a covalent sigma-bond and N-N=145.1/145.4(3) pm. The terminal B--N distance 139.6/138.9(9) pm is consistent with a covalent sigma-bond augmented by a dative pi-bond. An open Y-shaped structure also corresponds to a minimum on the potential energy surface, but the energy is 18.7 kJ mol(-1) higher (MP2) and it is incompatible with the GED data.


Running on a treadmill: dynamic inhibition of APC/C by the spindle checkpoint.
During mitosis, the genome duplicated during S-phase is synchronously and accurately segregated to the two daughter cells. The spindle checkpoint prevents premature sister-chromatid separation and mitotic exit. The anaphase-promoting complex/cyclosome (APC/C) is a key target of the spindle checkpoint. Upon checkpoint activation, the mitotic checkpoint complex (MCC) containing Mad2, Bub3, Mad3/BubR1 and Cdc20 inhibits APC/C. Two independent studies in budding yeast have now shed light on the mechanism by which MCC inhibits APC/C. These studies indicate that Mad3 binds to the mitotic activator of APC/C Cdc20 using peptide motifs commonly found in APC/C substrates and thus competes with APC/C substrates for APC/CCdc20 binding. In addition, Mad3 binding to APC/CCdc20 induces Cdc20 ubiquitination by APC/C, leading to the dissociation of MCC. Meanwhile, two other studies have shown that a deubiquitinating enzyme is required for the spindle checkpoint whereas APC/C-dependent ubiquitination is needed for checkpoint inactivation. Collectively, these studies suggest a dynamic model for APC/CCdc20 regulation by MCC in which APC/C- and Mad3-dependent ubiquitination of Cdc20 constitutes a self-regulated switch that rapidly inactivates the spindle checkpoint upon correct chromosome attachment.


Mixture modeling of microarray gene expression data.
About 28% of genes appear to have an expression pattern that follows a mixture distribution. We use first- and second-order partial correlation coefficients to identify trios and quartets of non-sex-linked genes that are highly associated and that are also mixtures. We identified 18 trio and 35 quartet mixtures and evaluated their mixture distribution concordance. Concordance was defined as the proportion of observations that simultaneously fall in the component with the higher mean or simultaneously in the component with the lower mean based on their Bayesian posterior probabilities. These trios and quartets have a concordance rate greater than 80%. There are 33 genes involved in these trios and quartets. A factor analysis with varimax rotation identifies three gene groups based on their factor loadings. One group of 18 genes has a concordance rate of 56.7%, another group of 8 genes has a concordance rate of 60.8%, and a third group of 7 genes has a concordance rate of 69.6%. Each of these rates is highly significant, suggesting that there may be strong biological underpinnings for the mixture mechanisms of these genes. Bayesian factor screening confirms this hypothesis by identifying six single-nucleotide polymorphisms that are significantly associated with the expression phenotypes of the five most concordant genes in the first group.


[Diagnostic value of IgG, IgA, and IgM to Bordetella pertussis antigens in patients with pertussis].
Comparative study of IgG, IgA, and IgM levels to complex of antigens (CA) of vaccine strain No. 475 and separate antigens of Bordetella pertussis: pertussis toxin (PT), filamentous hemagglutinin (FHA), lypopolysaccharide (LPS), agglutinogens 1 (Aggl.1) and 2 (Aggl.2) was performed by ELISA in 80 patients with pertussis and 80 healthy vaccinated children. Antibodies to mentioned antigens were detected both in ill and healthy children but their levels were remarkably higher in patients. The most reliable serologic marker of pertussis was IgA, which were detected in the majority of patients. Detection rates of this class of antibodies to CA, PT, FHA, LPS, Aggl.1, and Aggl.2 were 91%, 77.5%, 69%, 59%, 80%, and 12%, respectively. Elevated levels of specific IgA were registered in 5% of healthy children. Obtained results showed high information value of detection of the IgA and IgG antibodies to CA, PT, FHA, and Aggl.1 using ELISA for pertussis diagnostics. Simplicity and economy of the CA obtainment allow to recommend CA-based ELISA for serologic diagnostics of pertussis.


Spike generation from dorsal roots and cutaneous afferents by hypoxia or hypercapnia in the rat in vivo.
The present study aimed at investigating the responsiveness of different parts of the primary afferent neurones to a brief hypoxia, hypercapnia or ischaemia under in vivo conditions. Action potentials were recorded in separate groups of anaesthetized rats from (i) the peripheral end of the central stump of the cut L3, L4 or L5 dorsal root (dorsal root preparation); (ii) the central end of the peripheral stump of the cut saphenous nerve (saphenous-receptor preparation); (iii) the distal end of a segment of the saphenous nerve cut at both ends (axon preparation). In paralysed animals interruption of artificial ventilation for 20-60 s elicited or increased the frequency of action potentials in both the dorsal root and saphenous-receptor preparations. Activation of these preparations was also achieved by inspiration of gas mixtures containing 10-0% oxygen (mixed with nitrogen) or 20-50% carbon dioxide (mixed with oxygen) which elicited in the blood a decrease in PO2 or an increase in PCO2 with a fall in pH. Occlusion of the femoral artery for 3 min also caused spike generation in the saphenous-receptor preparations with little alteration in blood pressure. All these stimuli failed to evoke action potentials in the axon preparations. Systemic (300 mg kg-1 s.c.) or perineural (2%) capsaicin pretreatment failed to inhibit the effect of hypoxia, hypercapnia or ischaemia, indicating a significant contribution of capsaicin-insensitive neurones to the responses. It is concluded that central and peripheral terminals but not axons of primary afferent neurones are excited by a brief hypoxia or hypercapnia and the peripheral terminals by a short local ischaemia as well. Excitation of central terminals by hypoxia or hypercapnia revealed in this way an antidromic activation of dorsal roots in response to natural chemical stimuli.


[Reduction of functional mobility and cognitive capacity in type 2 diabetes mellitus].
The aim of the present study was to evaluate the functional mobility and its relationship to cognitive ability in patients with type 2 diabetes (T2DM), age between 50 and 65 years and under 10 years of diagnosis. An observational, analytical and cross-sectional study, involving no diabetic and type 2 diabetic individuals with inadequate glycemic control, selected by convenience sampling. In both groups, were administered structured questionnaire and cognitive assessment with Mini-Mental State Examination (MMSE) and the clock drawing test (CDT), besides assessment of functional mobility by the Timed Up & Go (TUG). In TUG, DM2 patients presented a mean time of 11.27 seconds versus 9.52 seconds (p = 0.013). The association between cognitive decline and decrease of mobility was positive in individuals with T2DM (p = 0.037). In the subgroup that showed decrease of mobility and associated cognitive decline, 18% were patients with DM2 and 1.6% were individuals without T2DM (p < 0.01). Patients with T2DM presented worse functional mobility and cognitive performance, supporting the hypothesis that DM2 influence functional mobility and cognitive ability, regardless of neuropathic or vascular complications. These data suggest that hyperglycemia is an aggravating factor in the performance of activities requiring mental functions such as attention, working memory and orientation.


Prevalence of medical marijuana use in California, 2012.
The US Drug Enforcement Agency classifies marijuana as an illegal substance, yet in 22 states marijuana is legal for medicinal use. In 1996, California legalised the use of marijuana for medicinal purposes, but population-based data describing medical marijuana users in the state has not been available. Our aim was to examine the demographic differences between users and non-users of medical marijuana in California utilising population-based data. We used data from the California Behavioral Risk Factor Surveillance System 2012, an annual, random-digit-dial state-wide telephone survey that collects health data from a representative adult sample (n = 7525). Age-adjusted prevalence rates were estimated. Five percent of adults in California reported ever using medical marijuana, and most users believed that medical marijuana helped alleviate symptoms or treat a serious medical condition. Prevalence was similar when compared by gender, education and region. Prevalence of ever using medical marijuana was highest among white adults and younger adults ages 18-24 years, although use was reported by every racial/ethnic and age group examined in our study and ranged from 2% to 9%. Our study's results lend support to the idea that medical marijuana is used equally by many groups of people and is not exclusively used by any one specific group. As more states approve marijuana use for medical purposes, it is important to track medical marijuana use as a health-related behaviour and risk factor.


The role of enzyme activation state in limiting carbon assimilation under variable light conditions.
The mechanisms regulating transient photosynthesis by soybean (Glycine max) leaves were examined by comparing photosynthetic rates and carbon reduction cycle enzyme activities under flashing (saturating 1 s lightflecks separated by low photon flux density (PFD) periods of different durations) and continuous PFD. At the same mean PFD, the mean photosynthetic rates were reduced under flashing as compared to continuous light. However, as the duration of the low PFD period lengthened, the CO2 assimilation attributable to a lightfleck increased. This enhanced lightfleck CO2 assimilation was accounted for by a greater postillumination CO2 fixation occurring after the lightfleck. The induction state of photosynthesis, ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco), fructose 1,6-bisphosphatase (FBPase) and ribulose 5-phosphate kinase (Ru5P kinase) activities all responded similarly and were all lower under flashing as compared to constant PFD of the same integrated mean value. However, the fast phase of induction and FBPase and Ru5P kinase activities were reduced more than were the slow phase of induction and rubisco activity. This was consistent with the role of the former enzymes in the fast induction component that limited RuBP regeneration. Competition for reducing power between carbon metabolism and thioredoxin-mediated enzyme activation may have resulted in lower enzyme activation states and hence lower induction states under flashing than continuous PFD, especially at low lightfleck frequencies (low mean PFD).


[Association between the Gut Microbiome and Cognitive Function].
Dysregulation of the gut microbiome is associated with several life-threatening conditions, and might therefore represent a useful target for the prevention of dementia. However, the relationship between the gut microbiome and dementia has not yet been fully elucidated. Here, we recruited outpatients visiting our memory clinic to participate in this study. Information for patient demographics, various risk factors, and daily activities was collected, and cognitive function was assessed using neuropsychological tests and magnetic resonance imaging scans. Fecal samples were obtained, and the gut microbiome was assessed by terminal restriction fragment length polymorphism (T-RFLP) analysis, one of the most well-established and reliable 16S ribosomal RNA-based methods for classifying gut microbiota. Multivariable logistic regression models were used to identify factors independently associated with dementia and mild cognitive impairment. Graphical modelling was used to illustrate mutual associations. We analyzed 128 eligible patients (female: 59%, mean age: 74.2±8.7 years, mean Mini Mental State Examination score 24). Multivariable analyses showed that enterotype I and enterotype III bacteria were strongly associated with dementia, independent of traditional dementia biomarkers. Further studies investigating metabolites of gut microbes are needed to determine the mechanism underlying this association.


Detection of autoantibodies to ribonucleoproteins by counter-immunoelectropheresis, immunoblotting and RNA-immunoprecipitation: comparison of results.
Counterimmunoelectropheresis (CIE), immunoblotting (IB) and RNA-immunoprecipitation (RNA-IP) were compared for their potential to detect antibodies to U1RNP, Sm and SS-B in sera from 50 patients with SLE and 18 with MCTD. Anti-SS-B were detected in 6 SLE-sera (12%) and one MCTD-serum (6%) by all three techniques. Anti-Sm were present in 7 SLE-sera (14%) by RNA-IP and IB; 3 of these sera showed identity with a-Sm and 4 with a-U1RNP by CIE. Antibodies precipitating U1RNA were present in 8 SLE sera (16%); 4 of these sera were positive for anti-U1RNP by CIE (8%) and 4 sera recognized one or more of the U1RNA-associated proteins by IB (8%). All MCTD-sera were positive for anti-U1RNP by RNA-IP including one serum negative by IB and CIE. RNA-IP allowed the detection of antibodies precipitating U1-U2RNA in 4 sera; in IB, the U2RNA specific protein B" was recognized by 3 sera. RNA-IP appears to be a sensitive method for detecting anti-URNP's.


Actinobacillus vicugnae sp. nov., isolated from alpaca (Vicugna pacos).
Ten strains of an Actinobacillus-like organism were isolated from alpaca (Vicugna pacos) in the UK over a period of 5 years, with no known epidemiological linkages. The isolates are distinct, based on both phenotype and genotype, from any previously described Actinobacillus species. Molecular analysis, based on 16S rRNA, rpoB and infB gene sequences, placed the isolates as a novel, early branching, lineage within the currently recognised Actinobacillus sensu stricto. In agreement with the results of the single-gene analysis, average nucleotide identity values, based on whole genome sequences, showed very similar identities to a number of members of the Actinobacillus sensu stricto notably Actinobacillus equuli, Actinobacillus suis and Actinobacillus ureae. At least two phenotypic characteristics differentiate the alpaca isolates from other Actinobacillus sensu stricto species, and from taxa likely falling within this group but awaiting formal species description, with Actinobacillus anseriformium and A. equulisubsp. haemolyticus being the most closely related phenotypically. The alpaca isolates can be differentiated from A. anseriformium by production of β-galactosidase (ONPG) and acid from raffinose, and from A. equulisubsp. haemolyticus by production of acid from d-sorbitol and failure to produce acid from d-xylose. Isolates were obtained from multiple sites in alpaca including respiratory tract, alimentary tract and internal organs although further evidence is required to understand any pathogenic significance. Based on the results of characterization described here, it is proposed that the isolates constitute a novel species, Actinobacillus vicugnae sp. nov. The type strain is W1618T (LMG30745T NCTC14090T) isolated in the UK in 2012 from oesophageal ulceration in an alpaca (Vicugna pacos).


Dumbbell granulomatous abscess of the chest wall following needle biopsy of the pleura.
A 38-year-old woman who had a Cope needle biopsy of the pleura was treated for plural tuberculosis on the basis of a positive PPD-S skin test and presence of caseating granulomas in the pleural biopsy. Ten months later she developed a tender, subcutaneous nodule in the area of the previous needle biopsy. Surgical exploration revealed a dumbbell abscess through the chest wall communicating with an area of consolidation in the right middle lobe. En bloc surgical resection of the abscess and peripheral portion of the right middle lobe was curative, although all pathologic and cultural studies of the resected tissue were non-diagnostic.


[Clinical case of a combination of ulcerative colitis and alopecia areata].
Extraintestinal manifestations of inflammatory bowel diseases (IBD) are common in both ulcerative colitis and Crohn's disease. Dermatological manifestations have been reported in one-third of all extraintestinal manifestations of IBD. Erythema nodosum and pyoderma gangrenosum are the most frequently described in the literature. There is no data on the development of alopecia as a cutaneous manifestation of IBD in most of studies. A clinical case of combination of ulcerative colitis and alopecia in young woman is presented in the article. There are several possible causes of alopecia in patients with IBD, such as nutritional deficiencies, side effects of medications or autoimmune alopecia. Most often there is no correlation between the severity of clinical bowel manifestations and other organs involvement, and response to therapy. So the combination of alopecia with IBD, in particular ulcerative colitis, confirms the importance of autoimmune mechanisms in genesis of these diseases.


Evaluation with evoked and spontaneous electromyography during lumbar instrumentation: a prospective study.
The neuroanatomical structures that approximate the bony pedicles of the lumbar spine allow little room for technical error or compromise of the bone during pedicle screw insertion. Currently available neurophysiological monitoring techniques detect compromised bone and nerve root injury after it occurs. The purpose of this prospective study is to evaluate the reliability and efficacy of a unique neurophysiological monitoring technique. This technique provides immediate evaluation of pedicle cortical bone integrity in patients undergoing lumbar fusion with instrumentation by using electrified surgical instruments throughout the pedicle screw fusion procedure. Spontaneous electromyographic (EMG) activity was also monitored. Intraoperative evoked EMG stimulation was performed using a pedicle probe and feeler as monopolar stimulators during the insertion of 164 pedicle bone screws in 32 patients. The EMG response to subthreshold stimulation intensities indicated cortical bone compromise. Immediate and conclusive feedback via evoked EMG activity using stimulating pedicle probes in appropriate muscle groups was successful in identifying pedicle cortical bone compromise in four patients. One false-negative evoked EMG study was noted but was identified via spontaneous EMG activity. Intraoperative EMG monitoring alerted the surgeon that redirection of the pedicle probe or screw was necessary to avoid nerve root irritation or injury and served as an early warning system. Evoked EMG stimulation proved to be reliable and efficacious, especially when used in combination with spontaneous EMG. This technique may provide an added safeguard during implant placement procedures at centers where intraoperative neurophysiological monitoring is routinely performed.


Effect of chromium species on phytochemical and physiological parameters in Datura innoxia.
In soil, chromium can be found in two main valence states: hexavalent Cr(VI) and trivalent Cr(III). In this study, we investigated the impact of Cr on photosynthetic gas exchange, photosystem II (PSII) activity, Cr translocation and accumulation, proline content and alkaloids production, i.e. scopolamine and hyoscyamine, in Datura innoxia. Cr uptake was influenced by its oxidation state and its concentration in growth medium. The plant roots were determined as being the main organ of Cr accumulation. Cr(VI) was more toxic than Cr(III) as indicated by reduction in plant biomass and net photosynthesis. The stomatal conductance showed a similar trend to that of photosynthetic capacity. Cr(III) and Cr(VI) had a different impact on substomatal CO(2) concentration then Cr toxicity was related to its oxidation states. In plants stressed with a Cr(VI) excess, a down regulation of PSII activity was observed with an impairment of photochemical activity. Indeed, the maximum quantum yield of PSII (F(v)/F(m)), the quantum yield of PSII (PhiPSII) and the efficiency of excitation capture by open centers (F'(v)/F'(m)) decreased. Cr(III) had little effects on PSII primary photochemistry, whatever its form induces an increase of scopolamine content without changes in hyoscyamine content in leaves of D. innoxia. These results provide that chromium contamination can change the secondary metabolites composition of leaves, thereby, impacting the quality, safety and efficacy of natural plant products synthesized by D. innoxia plants.


Hunting income patterns among the Hadza: big game, common goods, foraging goals and the evolution of the human diet.
The assumption that large mammal hunting and scavenging are economically advantageous to hominid foragers is examined in the light of data collected among the Hadza of northern Tanzania. Hadza hunters disregard small prey in favour of larger forms (mean adult mass greater than or equal to 40 kg). Here we report experimental data showing that hunters would reduce their mean rates if they included small animals in the array they target. Still, daily variance in large animal hunting returns is high, and the risk of failure correspondingly great, significantly greater than that associated with small game hunting and trapping. Sharing large kills reduces the risk of meatless days for big game hunters, and obviates the problem of storing large amounts of meat. It may be unavoidable if large carcasses cannot be defended economically against the demands of other consumers. If so, then large prey are common goods. A hunter may gain no consumption advantage from his own big game acquisition efforts. We use Hadza data to model this 'collective action' problem, and find that an exclusive focus on large game with extensive sharing is not the optimal strategy for hunters concerned with maximizing their own chances of eating meat. Other explanations for the emergence and persistence of this practice must be considered.


A Bump-Hole Approach for Directed RNA Editing.
Molecules capable of directing changes to nucleic acid sequences are powerful tools for molecular biology and promising candidates for the therapeutic correction of disease-causing mutations. However, unwanted reactions at off-target sites complicate their use. Here we report selective combinations of mutant editing enzyme and directing oligonucleotide. Mutations in human ADAR2 (adenosine deaminase acting on RNA 2) that introduce aromatic amino acids at position 488 reduce background RNA editing. This residue is juxtaposed to the nucleobase that pairs with the editing site adenine, suggesting a steric clash for the bulky mutants. Replacing this nucleobase with a hydrogen atom removes the clash and restores editing activity. A crystal structure of the E488Y mutant bound to abasic site-containing RNA shows the accommodation of the tyrosine side chain. Finally, we demonstrate directed RNA editing in vitro and in human cells using mutant ADAR2 proteins and modified guide RNAs with reduced off-target activity.


A comparison between distal and proximal port device insertion in head and neck cancer.
To compare technical feasibility and complications of radiologically arm port device implantation using arm venography exclusively (API-Group B) with chest port placement using cephalic vein cutdown (CVC-Group A), in advanced consecutive head and neck cancer patients (HNP). Port device placement was attempted in 225 consecutive HNP. Decision for inclusion in Group A or B was made first by the availability of the surgeon/radiologist to perform the procedure, second by contraindications of each technique. Patient transfer from one group to the other was recorded as well as technical feasibility, complications and device specific duration in this retrospective study. Technical success was statistically higher in Arm Port Group (99.1%) compared to Chest Port Group (75.2%). Device specific duration rate of the whole population was 53% (95%CI) [0.47-0.60] at 6 months, 44.1% (95%CI) [24.4-37.8] at 12 months and 8% (95%CI) [4.4-14.5] at 24 months. Median follow-up was 5.55 months (range: 0.032-9.6] in Group A versus 5.90 months [range: 0.06-27.6] (p=ns) in Group B. Complication rate was 15.9% in Group A versus 8.9% in Group B corresponding to a complication rate per patient-implantation-days of 0.66/1000 patient-days (A) versus 0.42/1000 patient-days (B). Premature port device explantation rate was 4.4% (A) versus 5.4% (B). Axillary and subclavian venous thrombosis was the main complication and occurred in 12 Group A patients and three Group B patients. Venous thrombosis rate was 0.37/1000 patient-days (A) and 0.13/1000 patient-days (B) (p=0.03). A few data exist about device insertion in HNP in whom venous cervical access is contraindicated. This comparative study demonstrates that both implantation techniques are safe and effective. The higher technical success rate with 0% heavy sedation, the lower venous thrombosis rate in the API group, and the 5.3% (A-B) patient transfer rate argue in favour of arm port placement in HNP. Indications for API include patients with an ipsilateral major pectoralis-myocutaneous flap, with radiodermatitis, tumour recurrence in the neck and upper chest, or with respiratory impairment.


Chylopericardium following cardiac surgery.
Chylopericardium is one of the rarest complications of cardiac surgery. The authors report only the 19th case, that of a 12-year-old boy. The chylopericardium developed after an aortic valvotomy, even though the retropericardial portion of the mediastinum was entered only to pass a tape around the ascending aorta. Etiologic factors are directly or indirectly related to injury to the thoracic duct or tis tributaries. Treatment initially is conservative, consisting of adequate pericardial drainage and a low-fat, medium-chain triglyceride diet. If this fails, the appropriate operative procedure is carried out.


Single-bottle adhesives behave as permeable membranes after polymerization. I. In vivo evidence.
This study tested the hypothesis that single-bottle total-etch adhesives are effective in reducing dentine permeability under in vivo conditions. Crown preparations on vital human teeth were performed under local analgesia as part of the treatment plan for prosthetic rehabilitation. Four single-bottle adhesives (Single Bond, 3M ESPE; Excite DSC, Ivoclar Vivadent; Prime and Bond NT Dual-Cure, Dentsply DeTrey and One-Step, Bisco Inc.) were applied to the cut dentine after acid-etching. Polyvinyl siloxane impressions were taken, using an ultra-low viscosity impression material, of the smear layer-covered dentine before applying the adhesives to deep vital dentine, and after adhesive placement. Additional impressions were taken of the adhesive-sealed dentine following the removal of the provisional prostheses after a 7-10 day period. Epoxy resin replicas of the crown preparations were examined with scanning electron microscopy to evaluate the extent of dentinal fluid transudation during pre-bonded, immediately bonded and post-bonded periods. Dentinal fluid transudation from localised areas that were close to the dental pulp was universally observed from all epoxy resin replicas, irrespective of the adhesive employed. The transudation of dentinal fluid from the control smear layer-covered dentine of each crown preparation was comparatively mild when compared to the extent that was observed after total-etching and application of the single-bottle adhesive. Dentinal fluid droplets were specifically located over the surface of the adhesive layer. Continuous transudation of dentinal fluid occurred even after the removal of the provisional prostheses, and was considerably more profuse in subjects who opted for the fitting of the permanent prostheses without the use of a local anaesthetic solution. Single-bottle adhesives, because of their lack of a comparatively more hydrophobic bonding resin layer, behave as permeable membranes after polymerisation. They permit the continuous transudation of dentinal fluid and do not provide a hermetic seal in vital deep dentine. Although the relatively slow rate of diffusion of dentinal fluid is unlikely to result in post-operative cold sensitivity, it may interfere with the optimal polymerisation of dual-cured or auto-cured composites or resin cements in both direct and indirect restorations.


Four-dimensional Printing of Liquid Crystal Elastomers.
Three-dimensional structures capable of reversible changes in shape, i.e., four-dimensional-printed structures, may enable new generations of soft robotics, implantable medical devices, and consumer products. Here, thermally responsive liquid crystal elastomers (LCEs) are direct-write printed into 3D structures with a controlled molecular order. Molecular order is locally programmed by controlling the print path used to build the 3D object, and this order controls the stimulus response. Each aligned LCE filament undergoes 40% reversible contraction along the print direction on heating. By printing objects with controlled geometry and stimulus response, magnified shape transformations, for example, volumetric contractions or rapid, repetitive snap-through transitions, are realized.


[Latest indications for hanging manoeuvre in liver surgery].
By definition the liver hanging manoeuvre (LHM) means that a slip is passed between the liver parenchyma and the inferior vena cava. It was first published by Belghiti in 200l, and several changes in the indication as well as in the method have been published since then. In parallel, the anatomical and histological basis has been clarified for LHM, too. According to general consensus LHM increases safety and radicality of liver surgery. Initially LHM was applied for removal of huge tumours infiltrating the diaphragm. Authors worked out two modifications for LHM. Tumours / primary or secondary / in segment IVA are sometimes located in close proximity to the median hepatic vein and inferior vena cava , and the resectability of these tumors can determined by the hanging manoeuvre. Tumors in segment VII can be removed by partial resection of vena cava facilitated by LHM. Four patients with LHM are discussed, and based on this limited experience as well as the latest observations from relevant literature the authors claim that LHM increases the safety of resections from segment IVA and VII. Vascular infiltration of the vena cava is always a technical challenge, which can be suspected on preoperative imaging modalities, but in borderline cases only the intraoperative ultrasound and surgical judgment together with LHM would lead to the exact diagnosis and makes the resection possible.


First Report of Dieback Caused by Neofusicoccum batangarum in Cashew in Guinea-Bissau.
Cashew (Anacardium occidentale L.) is a cash crop with a highly significant economic importance in West Africa, particularly in Guinea-Bissau (Monteiro et al. 2015, 2017). In October 2018, dieback-like symptoms such as wilt and necrosis of apical shoots were observed in 10 % of the cashew trees grown in a 100 plant-orchard in Bolama Island at Bijagós archipelago, Guinea-Bissau. Six symptomatic apical shoots from individual plants were collected for fungal isolation and identification. Tissue pieces (3 × 2 mm) from healthy to diseased margins were surface sterilized with 1 % sodium hypochlorite, washed twice with sterilized water, placed on potato dextrose agar (PDA, Difco® Laboratories) supplemented with potassium thiocyanate (50 µg/ml), and incubated at 24 ± 1 °C in the dark for 7 days. Four fungal colonies were isolated (67 %) and purified through hyphal tips removal, displaying rapid growth rate, and aerial mycelia that initially was white, turning later to dark greenish on PDA. Pycnidia produced on 1.5 % water agar and sterilized pine needles (± 25ºC; near-UV light) were solitary, covered by mycelium, obpyriform to ampulliform (152.5 ± 41.6 × 135.2 ± 30.8 μm, n = 30). Conidia were unicellular, hyaline, smooth, fusoid to ovoid, thin-walled, measuring 16.21 ± 1.52 × 5.84 ± 0.66 μm (n = 50, L/W 2.8). Such morphological features are characteristic of Neofusicoccum spp. (Phillips et al. 2013). For molecular identification, genomic DNA was extracted from a representative isolate GB160 and partial regions of ribosomal internal transcriber spacer (ITS) (ITS1/ITS4; White et al. 1990), translation elongation factor 1-α (EF1-α) (EF1-688F/EF1-1251R; Alves et al. 2008) and β-tubulin (β-tub) (Bt2a/Bt2b; Glass and Donaldson 1995) genes were amplified as previously described, respectively, with BSA (50 mg/ml). Amplicons were sequenced and deposited in GenBank (ITS, MN952993; EF1-α, MN952204; β-tub, MN952208). BLAST analysis of ITS, EF1-α and β-tub gene sequences showed 100 % identity with Neofusicoccum batangarum reference strain CBS124923 (FJ900608, FJ900654, FJ900635, respectively). Maximum-likelihood and Bayesian Inference analyses from the concatenated dataset placed GB160 isolate within the N. batangarum cluster (BS = 72 %; PP = 0.95). For pathogenicity assessment, 3-month-old cashew "Caju di Terra" plants (n = 8) grown in a greenhouse under controlled conditions were inoculated following a randomized block design as described by Lima et al. (2013). Briefly, 3 mm diam. stem tissue bark was removed and replaced with a 3 mm diameter PDA plug retrieved from the colony margin. Inoculation wound was covered with sterilized wet cotton and sealed with parafilm. Eight control plants were only treated with PDA plugs and the wound covered and sealed as described. After 15 days, all inoculated plants displayed similar symptoms to those observed in the field, and vascular lesions (10.8 ± 4.0 cm), whereas control plants remained symptomless. Koch's postulates were fulfilled by successful re-isolation of the pathogen from all inoculated stems and identification by morphology and gene sequencing. N. batangarum was identified associated with Anacardium spp. in Brazil (Netto et al. 2017) and recently reported as causing grapevine dieback in Brazil (Rêgo et al. 2020). To our knowledge, this is the first report of N. batangarum causing cashew dieback in Guinea-Bissau and West Africa. Occurrence of this disease may represent a significant impact for cashew production since this crop is the major agricultural commodity in Guinea-Bissau.


Endothelin-1-Induced Cell Hypertrophy in Cardiomyocytes is Improved by Fenofibrate: Possible Roles of Adiponectin.
Previous studies demonstrated that endothelin-1 (ET-1) can significantly increase the cell size and stimulate adiponectin expression in cultured human cardiomyocytes (HCM). The aim of the present study was to investigate the effects of fenofibrate, a peroxisome proliferator-activated receptor-α (PPARα) activator, on cell hypertrophy and adiponectin expression in vitro and in a rat model of daunorubicin-induced cardiomyopathy. The cultured human cardiomyocytes (HCM) were stimulated with or without ET-1. The cell size and the protein expressions of PPARα and adiponectin were tested by confocal Immunofluorescence study and Western blot, respectively. To study the effects of PPARα activation on ET-1-induced cell hypertrophy and adiponectin protein synthesis, HCM were pretreated with fenofibrate or small interfering RNA (siRNA) of PPARα. Echocardiographic parameters were measured and immunohistochemistry study of myocardial adiponectin expression was conducted in the in vivo study. ET-1 significantly increased the cell size, dose-dependently suppressed the expression of PPARα, and enhanced the expression of adiponectin; whereas, such an increase of cell size and enhancement of adiponectin expression were inhibited by the pre-treatment with fenofibrate. Addition of siRNA of PPARα abolished the effects of fenofibrate. Moreover, we found that fenofibrate treatment can significantly improve the left ventricular function and reverse the myocardial expression of adiponectin. Our study shows that fenofibrate may protect against ET-1-induced cardiomyocyte hypertrophy and enhanced adiponectin expression through modulation of PPARα expression in vitro and limitation of daunorubicin cardiotoxicity in vivo, suggesting a novel mechanistic insight into the role of PPARα and adiponectin in cardiac hypertrophy and heart failure.


In Situ Radiographic Investigation of (De)Lithiation Mechanisms in a Tin-Electrode Lithium-Ion Battery.
The lithiation and delithiation mechanisms of multiple-Sn particles in a customized flat radiography cell were investigated by in situ synchrotron radiography. For the first time, four (de)lithiation phenomena in a Sn-electrode battery system are highlighted: 1) the (de)lithiation behavior varies between different Sn particles, 2) the time required to lithiate individual Sn particles is markedly different from the time needed to discharge the complete battery, 3) electrochemical deactivation of originally electrochemically active particles is reported, and 4) a change of electrochemical behavior of individual particles during cycling is found and explained by dynamic changes of (de)lithiation pathways amongst particles within the electrode. These unexpected findings fundamentaly expand the understanding of the underlying (de)lithiation mechanisms inside commercial lithium-ion batteries (LIBs) and would open new design principles for high-performance next-generation LIBs.


Proportional mortality ratios among Korean immigrants to New York City, 1986-1990.
About 13,000 people immigrated to New York City from the Republic of Korea between 1986 and 1990, creating an important ethnic minority. Using US-born Whites as a reference, age-adjusted proportional mortality ratios were computed for 314 men and 248 women of Korean ancestry born abroad who died in New York City in 1986-90. Males had a significant excess of viral hepatitis, cancer, stroke, and external causes: accidents, suicide, and homicide. They had significantly reduced proportional rates of AIDS and heart disease. Mortality patterns were similar for Korean women, who had significantly increased proportional rates of stroke and accidents, and reduced heart disease. Stomach and liver cancers were significantly elevated in both sexes, while female breast cancer was low. There were two male and one female tuberculosis deaths (4 to 7 fold increase), and one Korean woman died of childbirth complications (59-fold increase). Except for violent deaths, these observations resemble known mortality patterns in Korea.


[Peptide and amino acid composition of different protein bases for culture media].
The enzymatic hydrolysates under study, obtained from different raw materials, have been shown to contain a great variety of peptides with different molecular weight. The highest content of fractions with a molecular weight of 2000 D has been observed in enzymatic meat and casein hydrolysates manufactures in the GDR. Low-molecular fractions (100-200 D) prevail in amino peptide. A great variety of peptides with different molecular weight is observed in Hottinger's meat hydrolysate and in blood clot hydrolysate obtained from the blood of laboratory animals. All peptide fractions have been shown to contain a wide spectrum of free amino acids. These data on the peptide and amino acid composition of different protein bases facilitate their rational use of microbiological culture media.


Metagenomic next-generation sequencing for diagnosis of infectious encephalitis and meningitis: a retrospective study of 90 patients.
Infections of the central nervous system (CNS) are potentially life-threatening and can cause serious morbidity. We evaluated the clinical value of metagenomic next-generation sequencing (mNGS) in the diagnosis of infectious encephalitis and meningitis and explored the factors affecting the results of mNGS. Patients with suspected cases of encephalitis or meningitis who presented in Northern Jiangsu People's Hospital from 1 March 2018 to 30 September 2022 were collected. Demographic, historical, and clinical information were obtained, and cerebrospinal fluid (CSF) samples were treated with mNGS. The pathogen was identified using National Center for Biotechnology Information (NCBI) GenBank sequence data. Ninety-six patients were screened and finally 90 subjects enrolled. Of the 90 enrolled cases, 67 (74.4%) were diagnosed with central nervous system infections, which included 48 cases (71.6%) of viral infection, 11 (12.2%) of bacterial infection, 5 (7.5%) of mycobacterium tuberculosis, 2 (3.0%) of fungal infection, and 1 (1.5%) of rickettsia infection. From these cases, mNGS identified 40 (44.4%) true-positive cases, 3 (3.3%) false-positive case, 22 (24.4%) true-negative cases, and 25 (27.8%) false-negative cases. The sensitivity and specificity of mNGS were 61.5% and 88%, respectively. mNGS of CSF could show a higher positive rate in patients with marked CSF abnormalities, including elevated protein concentrations and monocyte counts. mNGS of CSF is an effective method for detecting infectious encephalitis and meningitis, and the results should be analyzed combined with conventional microbiological testing results.


Ephrin-B2 reverse signaling is required for axon pathfinding and cardiac valve formation but not early vascular development.
Vascular development begins with the formation of a primary vascular plexus that is rapidly remodeled by angiogenesis into the interconnected branched patterns characteristic of mature vasculature. Several receptor tyrosine kinases and their ligands have been implicated to control early development of the vascular system. These include the vascular endothelial growth factor receptors (VEGFR-1 and VEGFR-2) that bind VEGF, the Tie-1 and Tie-2 receptors that bind the angiopoietins, and the EphB4 receptor that binds the membrane-anchored ligand ephrin-B2. Targeted mutations in the mouse germline have revealed essential functions for these molecules in vascular development. In particular, protein-null mutations that delete either EphB4 or ephrin-B2 from the mouse have been shown to result in early embryonic lethality due to failed angiogenic remodeling. The venous expression of EphB4 and arterial expression of ephrin-B2 has lead to the speculation that the interaction of these two molecules leads to bidirectional signaling into both the receptor-expressing cell and the ligand-expressing cell, and that both forward and reverse signals are required for proper development of blood vessels in the embryo. Indeed, targeted removal of the ephrin-B2 carboxy-terminal cytoplasmic tail by another group was shown to perturb vascular development and result in the same early embryonic lethality as the null mutation, leading the authors to propose that ephrin-B2 reverse signaling directs early angiogenic remodeling of the primary vascular plexus [Cell 104 (2001) 57]. However, we show here that the carboxy-terminal cytoplasmic domain of ephrin-B2, and hence reverse signaling, is not required during early vascular development, but it is necessary for neonatal survival and functions later in cardiovascular development in the maturation of cardiac valve leaflets. We further show that ephrin-B2 reverse signaling is required for the pathfinding of axons that form the posterior tract of the anterior commissure. Our results thus indicate that ephrin-B2 functions in the early embryo as a typical instructive ligand to stimulate EphB4 receptor forward signaling during angiogenic remodeling and that later in embryonic development ephrin-B2 functions as a receptor to transduce reverse signals involved in cardiac valve maturation and axon pathfinding.


Relative effectiveness of adjunctive risperidone on manic and depressive symptoms in mixed mania.
Mixed states are common and severe manifestations of bipolar disorder, with limited data on the differential efficacy of treatments on depressive and manic symptoms. This study assessed the effectiveness of open-label adjunctive risperidone in achieving sustained effectiveness in patients with mixed mania, with a specific focus on the differential benefits on manic and depressive symptomatology. Forty patients with bipolar disorder I, currently in a mixed manic episode, were treated with adjunctive risperidone. Behavioral measures at baseline and weeks 1, 2, 4, 8, 12, 16, and 20 were assessed using the following scales: the Young Mania Rating Scale, the Montgomery Asberg Depression Rating Scale (MADRS), and the Global Assessment Scale. The primary outcome measure was the proportion of patients who attained a sustained response on either depressive or manic symptomatology, defined as at least 50% reduction from the baseline on the Montgomery Asberg Depression Rating Scale or the Young Mania Rating Scale, maintained over at least 8 weeks without subsequent relapse during the 20-week trial. A significantly higher proportion of patients achieved a sustained response for mania than depression, 16/40 versus 6/40, respectively (McNemar's χ² 8.33, P=0.004). Higher elevated mood at baseline and lower apparent sadness (P<0.016) each predicted a sustained response for mania (P<0.0001). Mixed manic patients who were treated with risperidone adjunctive to mood stabilizer/s for 20 weeks were significantly more likely to achieve a sustained response for manic than for depressive symptomatology.


Signal peptide peptidase-mediated nuclear localization of heme oxygenase-1 promotes cancer cell proliferation and invasion independent of its enzymatic activity.
Heme oxygenase-1 (HO-1) is a heme-degrading enzyme anchored in the endoplasmic reticulum by a carboxyl-terminal transmembrane segment (TMS). HO-1 is highly expressed in various cancers and its nuclear localization is associated with the progression of some cancers. Nevertheless, the mechanism underlying HO-1 nuclear translocation and its pathological significance remain elusive. Here we show that the signal peptide peptidase (SPP) catalyzes the intramembrane cleavage of HO-1. Coexpression of HO-1 with wild-type SPP, but not a dominant-negative SPP, promoted the nuclear localization of HO-1 in cells. Mass spectrometry analysis of cytosolic HO-1 isolated from HeLa cells overexpressing HO-1 and SPP revealed two adjacent intramembrane cleavage sites located after S275 and F276 within the TMS. Mutations of S275F276 to A275L276 significantly hindered SPP-mediated HO-1 cleavage and nuclear localization. Nuclear HO-1 was detected in A549 and DU145 cancer cell lines expressing high levels of endogenous HO-1 and SPP. SPP knockdown or inhibition significantly reduced nuclear HO-1 localization in A549 and DU145 cells. The positive nuclear HO-1 stain was also evident in lung cancer tissues expressing high levels of HO-1 and SPP. Overexpression of a truncated HO-1 (t-HO-1) lacking the TMS in HeLa and H1299 cells promoted cell proliferation and migration/invasion. The effect of t-HO-1 was not affected by a mutation in the catalytic site. However, blockade of t-HO-1 nuclear localization abolished t-HO-1-mediated effect. The tumorigenic effect of t-HO-1 was also demonstrated in the mouse model. These findings disclose that SPP-mediated intramembrane cleavage of HO-1 promotes HO-1 nuclear localization and cancer progression independent of HO-1 enzymatic activity.


SET7/9 catalytic mutants reveal the role of active site water molecules in lysine multiple methylation.
SET domain lysine methyltransferases (KMTs) methylate specific lysine residues in histone and non-histone substrates. These enzymes also display product specificity by catalyzing distinct degrees of methylation of the lysine ε-amino group. To elucidate the molecular mechanism underlying this specificity, we have characterized the Y245A and Y305F mutants of the human KMT SET7/9 (also known as KMT7) that alter its product specificity from a monomethyltransferase to a di- and a trimethyltransferase, respectively. Crystal structures of these mutants in complex with peptides bearing unmodified, mono-, di-, and trimethylated lysines illustrate the roles of active site water molecules in aligning the lysine ε-amino group for methyl transfer with S-adenosylmethionine. Displacement or dissociation of these solvent molecules enlarges the diameter of the active site, accommodating the increasing size of the methylated ε-amino group during successive methyl transfer reactions. Together, these results furnish new insights into the roles of active site water molecules in modulating lysine multiple methylation by SET domain KMTs and provide the first molecular snapshots of the mono-, di-, and trimethyl transfer reactions catalyzed by these enzymes.


R on T Phenomenon and Long QTc Syndrome due to Moxifloxacin in a Healthy Female.
Long QT syndrome is characterized by prolongation of the corrected QT interval and is associated with fatal arrhythmias. R on T phenomenon is the coincidence of a premature ventricular complex with a T wave and may result to syncope and sudden cardiac death. Here we present a case of a 59-year-old woman with no previous history of medication, receiving moxifloxacin for community-acquired pneumonia. She was admitted complaining for thoracic pain and shortness of breath. Electrocardiogram showed both R on T phenomenon and QT prolongation. After discontinuation of the antibiotic symptoms and electrocardiography features were resolved. Physicians should always be vigilant about this rare and potentially fatal adverse drug reaction especially in high risk patients. General practitioners in particular being the first point of access in care have to carefully assess and detect high risk patients before the administration of moxifloxacin.


Bub1 is activated by the protein kinase p90(Rsk) during Xenopus oocyte maturation.
The kinetochore attachment (spindle assembly) checkpoint arrests cells in metaphase to prevent exit from mitosis until all the chromosomes are aligned properly at the metaphase plate. The checkpoint operates by preventing activation of the anaphase-promoting complex (APC), which triggers anaphase by degrading mitotic cyclins and other proteins. This checkpoint is active during normal mitosis and upon experimental disruption of the mitotic spindle. In yeast, the serine/threonine protein kinase Bub1 and the WD-repeat protein Bub3 are elements of a signal transduction cascade that regulates the kinetochore attachment checkpoint. In mammalian cells, activated MAPK is present on kinetochores during mitosis and activity is upregulated by the spindle assembly checkpoint. In vertebrate unfertilized eggs, a special form of meiotic metaphase arrest by cytostatic factor (CSF) is mediated by MAPK activation of the protein kinase p90(Rsk), which leads to inhibition of the APC. However, it is not known whether CSF-dependent metaphase arrest caused by p90(Rsk) involves components of the spindle assembly checkpoint. xBub1 is present in resting oocytes and its protein level increases slightly during oocyte maturation and early embryogenesis. In Xenopus oocytes, Bub1 is localized to kinetochores during both meiosis I and meiosis II, and the electrophoretic mobility of Bub1 upon SDS-PAGE decreases during meiosis I, reflecting phosphorylation and activation of the enzyme. The activation of Bub1 can be induced in interphase egg extracts by selective stimulation of the MAPK pathway by c-Mos, a MAPKKK. In oocytes treated with the MEK1 inhibitor U0126, the MAPK pathway does not become activated, and Bub1 remains in its low-activity, unshifted form. Injection of a constitutively active target of MAPK, the protein kinase p90(Rsk), restores the activation of Bub1 in the presence of U0126. Moreover, purified p90(Rsk) phosphorylates Bub1 in vitro and increases its protein kinase activity. Bub1, an upstream component of the kinetochore attachment checkpoint, is activated during meiosis in Xenopus in a MAPK-dependent manner. Moreover, a single substrate of MAPK, p90(Rsk), is sufficient to activate Bub1 in vitro and in vivo. These results indicate that in vertebrate eggs, kinetochore attachment/spindle assembly checkpoint proteins, including Bub1, are downstream of p90(Rsk) and may be effectors of APC inhibition and CSF-dependent metaphase arrest by p90(Rsk).


Characterization of variant neuroblastoma clones with missing or altered sodium channels.
Variant neuroblastoma cell clones were selected for resistance to the cytotoxic effects of neurotoxins that cause persistent activation of sodium channels. Three classes of variant clones were obtained: sodium channel-deficient clones, which have markedly reduced numbers of functional sodium channels; scorpion toxin-resistant clones, which have sodium channels with an altered interaction with scorpion toxin; and parental-type clones, which have functional sodium channels similar to the ones from N18 cells but have other heritable alterations that confer toxin resistance. The frequency of conversion to all three variant phenotypes was enhanced by treatment with the missense mutagen N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), suggesting that all three variant phenotypes are the result of mutational events. Incorporation of [35S]methionine into the alpha-subunit of the sodium channel (Mr = 270,000; pI = 5.8 +/- 0.2) was studied in normal and variant clones by affinity chromatography on wheat germ agglutinin/Sepharose followed by analysis of labeled polypeptides by 1- and 2-dimensional gel electrophoresis. Sodium channel-deficient clones do not synthesize the alpha-subunit of the sodium channel, suggesting that mutations in these clones block expression of the gene for this protein subunit. The scorpion toxin-resistant clone LV10 synthesizes an alpha-subunit which has a molecular weight and pI similar to those of the parental clones within the resolution of the methods used.


Increased risk for distant metastasis in patients with familial early-stage breast cancer and high EZH2 expression.
The identification of women with early-stage breast cancer who will develop distant metastasis may improve clinical management. The transcriptional regulator Enhancer of Zeste-2 (EZH2) is overexpressed in invasive breast carcinoma compared with benign breast tissues, with maximal expression in breast cancer metastasis. In this article, our purpose was to investigate the performance of EZH2 protein detection as a predictor of metastasis in women with early-stage breast cancer, which is unknown. We developed a cohort of 480 women with stage I-IIA breast cancer diagnosed between 1996 and 2002 and recorded detailed sociodemographic, clinical, and pathological information. Tumors were histologically characterized and arrayed in tissue microarrays containing 1,443 samples. The nuclear EZH2 expression was investigated by immunohistochemistry and was scored as 1-2 (negative and weak) or 3-4 (moderate and strong) using a validated scoring schema. Scores 1-2 were considered low EZH2; scores 3-4 were considered high EZH2. In this study, we found that after a median follow up of 9 years (range 0.04-14.5 years) 46 of 480 patients (9.6%) developed distant metastasis. High EZH2 was associated with larger size, high histological grade, negative hormone receptors, and first degree family history of breast and/or ovarian carcinoma. While EZH2 could not predict survival in the entire cohort, high EZH2 was a predictor of disease-specific survival in patients with early-stage disease and first degree family history (log rank P value 0.05). Importantly, in this group of patients, high EZH2 was an independent predictor of distant metastasis up to 15 years after primary carcinoma diagnosis (hazard ratio 6.58, 95% CI: 1.40-30.89, P = 0.016) providing survival information above and beyond currently used prognosticators. In conclusion, EZH2 may be a useful biomarker of long-term metastatic risk in women with familial early-stage breast cancer, and warrant further validation studies.


Sexting and high sexual risk-taking behaviours among school youth in northern Ethiopia: estimating using prevalence ratio.
Increasing access to digital technology to young people in low-income settings has greatly influenced their porngraphy viewing and sexting, receiving and/or sending of sexual explicit materials via electronic devices. These change the sexual communication and behaviour of the young population. However, evidence to attest this change is not available in our setting. Thus, this study examined the relationship of high sexual risk-taking behaviour with sexting and pornography viewing among school youth in Ethiopia. A cross-sectional study was conducted from March to April 2015 by selecting school youth using a multistage sampling procedure. Data were collected using a pre-validated anonymous facilitator-guided self-administered questionnaire. Poisson regression was run to calculate adjusted prevalence ratio with its 95% confidence intervals. All differences were considered as significant for p values ≤0.05. In total, 5924 questionnaires were distributed, and 5306 (89.57%) school youth responded in full to questions related to outcome variables. Of these respondents, 1220 (22.99%; 95% CI 19.45 to 26.96) were involved in high sexual risk-taking behaviour; 1769 (33.37%; 95% CI 30.52 to 36.35) had experienced sexting and 2679 (50.26%; 95% CI 46.92 to 53.61) were viewing pornography. The proportion of high sexual risk-taking behaviour was three-fold among pornography viewers (adjusted prevalence ratio (APR) 95% CI 3.02 (2.52 to 3.62)) and two-fold among sexters (APR 95% CI 2.48 (1.88 to 3.27)) as compared with their counterparts. Exposure to sexually explicit materials via communication technology is associated with increased high sexual risk-taking behaviour among school youth in northern Ethiopia. Considering these emerged predictors of sexual behaviours in our sexual education programmes, further research in this area is essential.


Ecological plant epigenetics: Evidence from model and non-model species, and the way forward.
Growing evidence shows that epigenetic mechanisms contribute to complex traits, with implications across many fields of biology. In plant ecology, recent studies have attempted to merge ecological experiments with epigenetic analyses to elucidate the contribution of epigenetics to plant phenotypes, stress responses, adaptation to habitat, and range distributions. While there has been some progress in revealing the role of epigenetics in ecological processes, studies with non-model species have so far been limited to describing broad patterns based on anonymous markers of DNA methylation. In contrast, studies with model species have benefited from powerful genomic resources, which contribute to a more mechanistic understanding but have limited ecological realism. Understanding the significance of epigenetics for plant ecology requires increased transfer of knowledge and methods from model species research to genomes of evolutionarily divergent species, and examination of responses to complex natural environments at a more mechanistic level. This requires transforming genomics tools specifically for studying non-model species, which is challenging given the large and often polyploid genomes of plants. Collaboration among molecular geneticists, ecologists and bioinformaticians promises to enhance our understanding of the mutual links between genome function and ecological processes.


A critical review of the current evidence examining whether resistance training improves time trial performance.
A number of reviews have concluded that resistance training is beneficial for improving sports performance despite the inclusion of studies which do not actually measure a performance outcome (i.e. a timed trial). The purpose of this review was to examine only those studies which would allow us to infer the benefits of resistance training on improving time trial performance. Of the nine studies meeting all inclusion criteria only three demonstrated an additive effect of adding resistance training to the current activity-specific training being performed. These three studies demonstrated improvements in either 5 or 10 km time trial among recreationally skilled athletes (i.e. non-elite level time). Previous reviews have included studies which did not include: (1) performance outcomes; (2) control groups; and/or (3) equal volumes of activity-specific exercise among the resistance training and control groups. Presently, there is little evidence that adding resistance exercise to a sport-specific training program will augment time trial performance. While it is difficult to perform such long-term studies assessing the effects of resistance training among time trial athletes, the statement that resistance training is efficacious for improving time trial performance should be tempered until sufficient evidence is presented to support such claims.


Effect of Microbial Biomass and Humic Acids on Abiotic and Biotic Magnetite Formation.
Magnetite (Fe3O4) is an environmentally ubiquitous mixed-valent iron (Fe) mineral, which can form via biotic or abiotic transformation of Fe(III) (oxyhydr)oxides such as ferrihydrite (Fh). It is currently unclear whether environmentally relevant biogenic Fh from Fe(II)-oxidizing bacteria, containing cell-derived organic matter, can transform to magnetite. We compared abiotic and biotic transformation: (1) abiogenic Fh (aFh); (2) abiogenic Fh coprecipitated with humic acids (aFh-HA); (3) biogenic Fh produced by phototrophic Fe(II)-oxidizer Rhodobacter ferrooxidans SW2 (bFh); and (4) biogenic Fh treated with bleach to remove biogenic organic matter (bFh-bleach). Abiotic or biotic transformation of Fh was promoted by Feaq2+ or Fe(III)-reducing bacteria. Feaq2+-catalyzed abiotic reaction with aFh and bFh-bleach led to complete transformation to magnetite. In contrast, aFh-HA only partially (68%) transformed to magnetite, and bFh (17%) transformed to goethite. We hypothesize that microbial biomass stabilized bFh against reaction with Feaq2+. All four Fh substrates were transformed into magnetite during biotic reduction, suggesting that Fh remains bioavailable even when associated with microbial biomass. Additionally, there were poorly ordered magnetic components detected in the biogenic end products for aFh and aFh-HA. Nevertheless, abiotic transformation was much faster than biotic transformation, implying that initial Feaq2+ concentration, passivation of Fh, and/or sequestration of Fe(II) by bacterial cells and associated biomass play major roles in the rate of magnetite formation from Fh. These results improve our understanding of factors influencing secondary mineralization of Fh in the environment.


MyoD stimulates RB promoter activity via the CREB/p300 nuclear transduction pathway.
The induction of RB gene transcription by MyoD is a key event in the process of skeletal muscle differentiation, because elevated levels of the retinoblastoma protein are essential for myoblast cell cycle arrest as well as for the terminal differentiation and survival of postmitotic myocytes. We previously showed that MyoD stimulates transcription from the RB promoter independently of direct binding to promoter sequences. Here we demonstrate that stimulation by MyoD requires a cyclic AMP-responsive element (CRE) in the RB promoter, bound by the transcription factor CREB in differentiating myoblasts. We also show that both the CREB protein level and the level of phosphorylation of the CREB protein at Ser-133 rapidly increase at the onset of muscle differentiation and that both remain high throughout the myogenic process. Biochemical and functional evidence indicates that in differentiating myoblasts, MyoD becomes associated with CREB and is targeted to the RB promoter CRE in a complex also containing the p300 transcriptional coactivator. The resulting multiprotein complex stimulates transcription from the RB promoter. These and other observations strongly suggest that MyoD functions by promoting the efficient recruitment of p300 by promoter-bound, phosphorylated CREB.


A Randomized, Double-Dummy, Emergency Department-Based Study of Greater Occipital Nerve Block With Bupivacaine vs Intravenous Metoclopramide for Treatment of Migraine.
Greater occipital nerve blocks (GONB) are used increasingly to treat acute migraine. We conducted a randomized controlled trial to determine whether GONB was as effective as intravenous metoclopramide for migraine. This was a double-dummy, double-blind, parallel-arm, non-inferiority study conducted in 2 emergency departments (EDs). Patients with migraine of moderate or severe intensity were randomized to receive bilateral GONB with each side administered 3 mL of bupivacaine 0.5% or metoclopramide 10 mg IV, the putative standard of care. The primary outcome was improvement in pain on a 0-10 scale between time 0 and 1 hour later. To reject the null hypothesis that metoclopramide would be more efficacious in relieving pain, we required that the lower limit of the 95% CI for the difference in pain improvement between those randomized to GONB vs those randomized to metoclopramide be >-1.3, a validated minimum clinically important difference. Secondary outcomes included sustained headache relief, defined as achieving and maintaining for 48 hours a headache level of mild or none without the use of additional analgesic medication, and the use of rescue medication in the ED. Over a 2.5-year study period, 1358 patients were screened for participation and 99 were randomized, 51 to GONB and 48 to metoclopramide. All of these patients were included in the primary analysis. Patients who received the GONB reported mean improvement of 5.0 (95% CI: 4.1, 5.8) while those who received metoclopramide reported a larger mean improvement of 6.1 (95% CI: 5.2, 6.9). The 95% CI for the between group difference of -1.1 was -2.3, 0.1. Sustained headache relief was reported by 11/51 (22%) GONB and 18/47 (38%) metoclopramide patients (95% CI for rounded difference of 17%: -1, 35%). Of the 51 GONB patients, 17 (33%) required rescue medication in the ED vs 8/48 (17%) metoclopramide patients (95% CI for rounded difference of 17%: 0, 33%). An adverse event was reported by 16/51 (31%) GONB patients and 18/48 (38%) metoclopramide patients (95% CI for (rounded) difference of 6%: -13, 25%). GONB with bupivacaine was not as efficacious as IV metoclopramide for the first-line treatment of migraine in the ED.


Child mortality associated with maternal HIV status: a retrospective analysis in Rwanda, 2005-2015.
Child mortality remains highest in regions of the world most affected by HIV/AIDS. The aim of this study was to assess child mortality rates in relation to maternal HIV status from 2005 to 2015, the period of rapid HIV treatment scale-up in Rwanda. We used data from the 2005, 2010 and 2015 Rwanda Demographic Health Surveys to derive under-2 mortality rates by survey year and mother's HIV status and to build a multivariable logistic regression model to establish the association of independent predictors of under-2 mortality stratified by mother's HIV status. In total, 12 010 live births were reported by mothers in the study period. Our findings show a higher mortality among children born to mothers with HIV compared with HIV negative mothers in 2005 (216.9 vs 100.7 per 1000 live births) and a significant reduction in mortality for both groups in 2015 (72.0 and 42.4 per 1000 live births, respectively). In the pooled reduced multivariable model, the odds of child mortality was higher among children born to mothers with HIV, (adjusted OR, AOR 2.09; 95% CI 1.57 to 2.78). The odds of child mortality were reduced in 2010 (AOR 0.69; 95% CI 0.59 to 0.81) and 2015 (AOR 0.35; 95% CI 0.28 to 0.44) compared with 2005. Other independent predictors of under-2 mortality included living in smaller families of 1-2 members (AOR 5.25; 95% CI 3.59 to 7.68), being twin (AOR 4.93; 95% CI 3.51 to 6.92) and being offspring from mothers not using contraceptives at the time of the survey (AOR 1.6; 95% CI 1.38 to 1.99). Higher education of mothers (completed primary school: (AOR 0.74; 95% CI 0.64 to 0.87) and secondary or higher education: (AOR 0.53; 95% CI 0.38 to 0.74)) was also associated with reduced child mortality. This study shows an important decline in under-2 child mortality among children born to both mothers with and without HIV in Rwanda over a 10-year span.


Protective effects of amino acids against gabexate mesilate-induced cell injury in porcine aorta endothelial cells.
Gabexate mesilate (GM), a serine protease inhibitor, often causes severe vascular injury. We previously reported that GM induced necrotic cell death via injury of the cell membrane in porcine aorta endothelial cells (PAECs). In the present study, we investigated the protective effects of amino acids against this GM-induced cell injury in PAECs. L-Cysteine (Cys), glycine (Gly), L-serine, L-glutamine (Gln), L-glutamate (Glu), L-proline, L-methionine, L-threonine, and L-isoleucine significantly inhibited the GM-induced decrease of cell viability. Gly showed the most potent effect among these amino acids. Gly, L-Cys, L-Glu, and L-Gln also inhibited the GM-induced increase in the number of necrotic cells stained by propidium iodide (PI). However, these amino acids had no effect on the GM-induced inhibition of trypsin activity. Strychnine, MK-801, or dichlorokynurenic acid did not affect the protective effect of Gly. Gly completely suppressed the GM-induced increase in PI uptake, which occurred immediately after exposure to GM. These findings suggest that Gly exerts protection against GM-induced cellular membrane injury, and several amino acids such as Gly may be useful for prophylaxis of the GM-induced severe vascular injury.


Existence and learning of oscillations in recurrent neural networks.
In this paper we study a particular class of -node recurrent neural networks (RNN's). In the 3-node case we use monotone dynamical systems theory to show, for a well-defined set of parameters, that, generically, every orbit of the RNN is asymptotic to a periodic orbit. Then we investigate whether RNN's of this class can adapt their internal parameters so as to "learn" and then replicate autonomously (in feedback) certain external periodic signals. Our learning algorithm is similar to identification algorithms in adaptive control theory. The main feature of the algorithm is that global exponential convergence of parameters is guaranteed. We also obtain partial convergence results in the -node case.


Crystalloid and perfluorochemical perfusates in an isolated working rabbit heart preparation.
Krebs-Henseleit buffer (KH) and a perfluorochemical (FC-43) were compared as perfusates in an isolated working rabbit heart preparation. Both perfusates were oxygenated in an identical manner using an infant bubble oxygenator. After 60 min of perfusion, no difference could be detected in the ratio of wet to dry heart weight between KH- and FC-43-perfused hearts (KH, 6.25 +/- 0.3; FC-43, 5.99 +/- 0.20). Left ventricular systolic pressure, maximal rate of left ventricular pressure rise, mean aortic systolic pressure, cardiac output, aortic flow, left ventricular power, and myocardial O2 consumption (MVO2) were significantly higher in FC-43-perfused hearts throughout the time of perfusion. However, there were no differences in resistance to cardiac output and heart rate. In KH- and FC-43-perfused hearts, MVO2 and left ventricular power were closely correlated (KH, r = 0.793; FC-43, r = 0.831). Significantly higher coronary flow of KH-perfused hearts could be attributed to the lower viscosity of KH (1.05 Pa . s) compared with FC-43 (1.91 Pa . s). Increased O2 extraction during KH perfusion could not compensate for low O2-carrying capacity of KH buffer (345 compared with 705 nmol O2 X ml-1 in FC-43 emulsion). A postischemic increase of coronary flow was observed only in FC-43-perfused hearts (28%). These results demonstrate a different response of perfused heart preparations to FC-43 and KH buffer.


Targeted CT imaging of human hepatocellular carcinoma using low-generation dendrimer-entrapped gold nanoparticles modified with lactobionic acid.
Development of cost-effective nanoscale contrast agents for targeted tumor computed tomography (CT) imaging remains a great challenge. Here, we report the synthesis of dendrimer-entrapped AuNPs (Au DENPs) using generation 2 poly(amidoamine) dendrimers pre-modified with fluorescein isothiocyanate via a thiourea linkage and lactobionic acid (LA) via a polyethylene glycol spacer as templates. The formed Au DENPs were characterized via different techniques and were used as a nanoprobe for targeted CT imaging of hepatocellular carcinoma (HCC). We show that the LA-modified Au DENPs with a mean Au core size of 1.8 nm are water-dispersible, colloidally stable under different temperatures (4-50 °C) and pH (5-8) conditions, and cytocompatible in the studied concentration range. Flow cytometry results reveal that the LA-Au DENPs are able to specifically target HepG2 cells (a human HCC cell line) overexpressing asialoglycoprotein receptors via a receptor-mediated targeting pathway. Importantly, the developed LA-Au DENPs can be used as a nanoprobe for targeted CT imaging of HepG2 cells in vitro and the xenografted tumor model in vivo. With the demonstrated organ compatibility, the developed LA-Au DENPs using low-generation dendrimers as templates can be a good candidate to be used as a highly efficient and cost-effective nanoprobe for targeted CT imaging of HCC.


Effect of adenosine triphosphate in renal ischemic injury: involvement of NF-kappaB.
Renal ischemic/reperfusion injury in vivo results in a significant increase of acute renal failure (ARF) and death. Nevertheless, there are many limitations in using in vivo models of renal ischemic injury to elucidate the detailed mechanisms of renal injury. Adenosine triphosphate (ATP), an extracellular signal, has been shown to be an important factor in regulation of epithelial cell function. Thus, the present study was performed to establish in vitro ischemic model using primary cultured rabbit renal proximal tubule cells (PTCs) and to examine the effect of ATP in this model. We established an in vitro model of ischemic injury, causing severe depletion of intracellular ATP by using the combination of a mitochondrial respiration inhibitor (antimycin A), non-metabolizable glucose analog (2-deoxyglucose), and calcium ionophore (A23187) in PTCs. Indeed, this ischemic injury significantly increased LDH release, a marker of structural damage, and ATP blocked ischemic injury-induced LDH release. 2-Methylthio-ATP and ATP-gamma-S (P2Y purinoceptor agonists) also blocked ischemic injury-induced LDH release, whereas AMP-CPP (P2X purinoceptor agonist) did not block it. In experiments to examine the relationship between ischemic injury and NF-kappaB activation, ischemic injury increased NF-kappaB translocation, DNA binding activity, and CAT activity. On the other hand, ATP, ATP-gamma-S, or 2-methylthio-ATP protected ischemic injury-induced NF-kappaB activation. These results suggest that the protective effect of ATP on ischemic injury is, in part, related to inhibition of NF-kappaB activation via P2Y receptor in PTCs.


Midpalatal suture maturation in 15- to 35-year-olds: morphological assessment in the coronal plane using CBCT-an exploratory study.
Rapid maxillary expansion (RME) is used in patients presenting transverse maxillary deficiency. However, RME may be unpredictable after late adolescence if the midpalatal sutural maturation stage (MPMS) is in late stage. Since MPMS evaluation is influenced by the expertise of the operator and image quality, this classification method could be complemented. Therefore, this study aimed to analyze the morphology of the midpalatal suture (MPS) and its surrounding bone in the coronal plane using cone beam computed tomography (CBCT) images of adolescents and young adults and to correlate the findings with their respective MPMS status. CBCT scans of 200 patients aged 15-35 years of both sexes were evaluated. MPS and surrounding bone tissue characteristics in the coronal and axial sections were analyzed, and MPMS was evaluated (A to E). Six categories were identified for the coronal evaluation, analyzing it in 2 standardized locations. Sutural morphology: I, hypodense sutural line limited by 2 hyper-dense para-sutural lines; II, hypodense sutural line; and III, suture not visible. The presence of para-sutural cancellous bone: CB0, compact bone; CB1, dense cancellous tissue; CB2, spaced cancellous bone. Midpalatal bone thickness (MBTh) was also measured. ANOVA-Bonferroni, chi-square, logistic regression and t-tests were used. MPMS increased with age. Sutural morphology I and CB0 bone were the most prevalent. Here we show that sutural morphology, para-sutural bone characteristics, and MPSM were significantly associated. The coronal evaluation of the MPS can complement the information from the MPSM assessment, allowing the identification of patent MPS even in the presence of late MPSM.


Testicular teratomas: an intersection of pluripotency, differentiation and cancer biology.
Teratomas represent a critical interface between stem cells, differentiation and tumorigenesis. These tumors are composed of cell types representing all three germ layers reflecting the pluripotent nature of their cell of origin. The study of these curious tumors became possible when Leroy Stevens identified the 129 mouse strain as a model of spontaneous testicular teratoma and later isolated a substrain carrying the Ter mutation, a potent modifier of tumor incidence. Early studies with 129 mice lead to the discovery of embryonal carcinoma (EC) cells which played a foundational role in the embryonic stem (ES) cell field and the study of pluripotency. The cells of origin of testicular teratomas are germ cells. During early development, primordial germ cells diverge from somatic differentiation and establish their pluripotent nature, maintaining or re-expressing core pluripotency genes; Oct4, Sox2 and Nanog. It is believed that a misregulation of male germ cell pluripotency plays a critical role in teratoma development. Several mouse models of teratoma development have now been identified, including a chromosome substitution strain, 129-Chr19(MOLF), conditional Dmrt1 and Pten alleles and the Ter mutation in the Dnd1 gene. However, it is still unknown what role somatic cells and/or physiology play in the sensitivity to teratoma development. These unusual tumors may hold the key to understanding how pluripotency is regulated in vivo.