Practical applications of genotypic surveys for forensic STR testing.
Legitimate genotype frequency estimation for multiallelic loci relies on component allele frequencies, as population surveys represent only a fraction of possible DNA profiles. Multilocus genotypes from two ethnic human populations, African American (n=195) and U.S. Caucasian (n=200), were compiled at 13 STR loci that are used worldwide in forensic investigation (D3S1358, vWA, FGA, D16S539, TH01, TPOX, CSF1PO, D8S1179, D21S11, D18S51, D5S818, D13S317, and D7S820). Sex-specific AmpFlSTR multiplexes provided stringent PCR-based STR typing specifically optimized for multicolor fluorescence detection. Heterozygosity at each STR locus ranged from 0.57 to 0.89 and encompassed from seven (TH01) to twenty-one (D21S11) alleles. Homozygosity tests, tests based on the distinct numbers of observed homozygous and heterozygous classes, log likelihood ratio tests, and exact tests assessed that the degree of divergence from theoretical Hardy-Weinberg proportions for all 13 STRs does not have practical consequence in genotype frequency estimation. Departures from linkage equilibrium, between loci, that imposed significance to forensic calculations were not indicated by observed variance of the number of heterozygous loci or Karlin interclass correlation tests. For forensic casework, reliable multilocus profile estimates may be obtained from the product of component genotype frequencies, each calculated through application of the Hardy-Weinberg equation to population database allele frequency estimates reported here. The average probability that two randomly selected, unrelated individuals possess an identical thirteen-locus DNA profile was one in 1.8x10(15) African Americans and one in 3.8x10(14) U.S. Caucasians.


Microsurgical treatment of parafalcine meningiomas: a retrospective study of 126 cases.
To discuss the diagnosis, operation methods, and clinical effects of parafalcine meningiomas. The clinical and preoperative imaging characteristics, operative methods, and effects of operations of 126 cases of parafalcine meningiomas were respectively discussed. G1 resection was achieved in 13 cases, G2 in 105 cases, G3 in four cases, and G4 in four cases, with no deaths. Among these, there were 16 patients with dyskinesia of the contralateral extremities after surgery, but they recovered after several months. In order to avoid postoperative complications, we consider it vital to analyze the patients' condition, the anatomy of venous drainage in by digital subtractional angiography, the relationship between tumor location and brain tissue according to MRI, and to remove the tumor in an adequately exposed surgical field.


Penicillin-induced generalized pustular psoriasis.
We describe a case of penicillin and penicillin-related drugs that induced severe generalized pustular psoriasis on several different occasions in a young patient known to have suffered from recalcitrant psoriasis since childhood.


Enzyme-linked immunosorbent assay for detection of Shiga toxin-producing Escherichia coli infection by antibodies to Escherichia coli secreted protein B in children with hemolytic uremic syndrome.
In order to detect immunoglobulin (Ig)A and IgG antibodies to Escherichia coli-secreted protein B in sera of children infected with Shiga toxin-producing Escherichia coli, an enzyme-linked immunosorbent assay was developed. The assay was tested using acute sera from 40 children with diarrhea-associated hemolytic uremic syndrome compared with 238 sera obtained from pediatric controls. Two cut-off values were used for children <5 (n=27) or > or =5 (n=13) years of age. Among the younger patients, 24 of 27 had IgA antibodies to Escherichia coli-secreted protein B (sensitivity, 89%; specificity, 98%) and 22 of 27 had IgG antibodies (sensitivity, 82%; specificity, 94%). Among the older patients, 13 of 13 had IgA antibodies (sensitivity, 100%; specificity, 96%) and 11 of 13 had IgG antibodies (sensitivity, 85%; specificity, 96%). This enzyme-linked immunosorbent assay detects Shiga-toxin-producing Escherichia coli independent of serogroup and could serve as a complementary assay for detection of infection.


Manipulation of a cation-π sandwich reveals conformational flexibility in phenylalanine hydroxylase.
Phenylalanine hydroxylase (PAH) is an allosteric enzyme that maintains phenylalanine (Phe) below neurotoxic levels; its failure results in phenylketonuria, an inborn error of amino acid metabolism. Wild type (WT) PAH equilibrates among resting-state (RS-PAH) and activated (A-PAH) conformations, whose equilibrium position depends upon allosteric Phe binding. The RS-PAH conformation of WT rat PAH (rPAH) contains a cation-π sandwich involving Phe80 that cannot exist in the A-PAH conformation. Phe80 variants F80A, F80D, F80L, and F80R were prepared and evaluated using native PAGE, size exclusion chromatography, ion exchange behavior, intrinsic protein fluorescence, enzyme kinetics, and limited proteolysis, each as a function of [Phe]. Like WT rPAH, F80A and F80D show allosteric activation by Phe while F80L and F80R are constitutively active. Maximal activity of all variants suggests relief of a rate-determining conformational change. Limited proteolysis of WT rPAH (minus Phe) reveals facile cleavage within a 4-helix bundle that is buried in the RS-PAH tetramer interface, reflecting dynamic dissociation of that tetramer. This cleavage is not seen for the Phe80 variants, which all show proteolytic hypersensitivity in a linker that repositions during the RS-PAH to A-PAH interchange. Hypersensitivity is corrected by addition of Phe such that all variants become like WT rPAH and achieve the A-PAH conformation. Thus, manipulation of Phe80 perturbs the conformational space sampled by PAH, increasing sampling of on-pathway intermediates in the RS-PAH and A-PAH interchange. The behavior of the Phe80 variants mimics that of disease-associated R68S and suggests a molecular basis for proteolytic susceptibility in PKU-associated human PAH variants.


An oral dsRNA delivery system based on chitosan induces G protein-coupled receptor kinase 2 gene silencing for Apolygus lucorum control.
RNA interference (RNAi) is recognized as a new and environmentally friendly pest control strategy due to its high specificity. However, the RNAi efficiency is relatively low in many sucking insect pests, such as Apolygus lucorum. Therefore, there is an urgent need to develop new and effective ways of dsRNA delivery. Bacterially expressed or T7 synthesized dsRNA targeting a G Protein-Coupled Receptor Kinase 2 gene was mixed with chitosan in a 1:2 ratio by mass. The size of the chitosan/dsRNA nanoparticles was 69 ± 12 nm, and the TEM and AFM images showed typical spherical or ellipsoidal structures. The chitosan nanoparticles protected the dsRNA from nuclease activity, and pH and temperature-dependent degradation, and the fluorescently-tagged nanoparticles were found to be stable on the surface of green bean plants (48 h) (Phaseolus vulgaris) and were absorbed by midgut epithelial cells and transported to hemolymph. Once fed to the A. lucorum nymph, chitosan/dsRNA could effectively inhibit the expression of the G protein-coupled receptor kinase 2 gene (70%), and led to significantly increase mortality (50%), reduced weight (26.54%) and a prolonged developmental period (8.04%). The feeding-based and chitosan-mediated dsRNA delivery method could be a new strategy for A. lucorum management, providing an effective tool for gene silencing of piercing-sucking insects.


n-Alkanols potentiate sodium channel inactivation in squid giant axons.
The effects of n-octanol and n-decanol on nerve membrane sodium channels were examined in internally perfused, voltage-clamped squid giant axons. Both n-octanol and n-decanol almost completely eliminated the residual sodium conductance at the end of 8-ms voltage steps. In contrast, peak sodium conductance was only partially reduced. This block of peak and residual sodium conductance was very reversible and seen with both internal and external alkanol application. The differential sensitivity of peak and residual conductance to alkanol treatment was eliminated after internal pronase treatment, suggesting that n-octanol and n-decanol enhance the normal inactivation mechanism rather than directly blocking channels in a time-dependent manner.


[Effect of electrical stimulation of the inferior central raphe nucleus on spontaneous and induced thalamic neuron activity in awake cats].
Using microelectrode technique in chronic experiments it was shown that electrical stimulation of the inferior raphe nucleus inhibited spontaneous activity of the thalamic neurons and that evoked by stimulation of radial nerves. Changes in the activity of the same neurons were absent in case of stimulation of the bulbar reticular formation.


A new series of cyclic amino acids as inhibitors of S-adenosyl L-methionine synthetase.
Optically active 3-amino-3-(tetrahydrofuranyl) carboxylic acid, 3-amino-3-(tetrahydrothienyl) carboxylic acid and their corresponding six membered ring analogues have been synthesised and examined as potential inhibitors of the enzyme S-adenosylmethionine (AdoMet) synthetase. The kinetic behaviour of these compounds was studied using recombinant rat liver AdoMet synthetase (alpha-isoform) fractionated from E. coli transformed with the plasmid pSSRL-T7N. All the compounds tested were competitive inhibitors of the enzyme with respect to L-methionine.


Clinical significance of eosinophilia and chronic inflammatory infiltrate in children's rectal biopsies.
The aim of the present study was to determine the clinical significance of an incidental finding of eosinophilia (EOS) and chronic inflammatory infiltrate (CINF) in rectal biopsies of children investigated for suspected Hirschsprung disease (HSCR). A retrospective study (2000-2010) of children incidentally found to have EOS and CINF was performed. HSCR cases were excluded. Presence of gastrointestinal (GI) symptoms and nutritional status (weight-for-age z score) were investigated and compared with a matched cohort with normal biopsy. Of 364 children undergoing rectal biopsy for suspected HSCR, 109 had confirmed HSCR, whereas 255 children had normal ganglia. Forty-four of 255 (17%) children had EOS and/or CINF incidentally reported and are the subject of the present study. In 13 of 44 (29%) children, the biopsy was performed neonatally. At follow-up (4.6 months [1-22]), 21 (48%) had food and/or milk allergy, 30 (68%) had constipation and/or other GI symptoms. There was no change in weight-for-age z score (P = 0.85) at follow-up and 8 (20%) had failure to thrive. Only 10 of 44 (P = 0.0001 vs patients with EOS and/or CINF) children with normal biopsy had persistent constipation at follow-up (9.7 months [0.5-84.7]) and 1 patient had atopy. Patients with normal biopsy exhibited an increase in weight-for-age z score at follow-up (P = 0.003) and only 3 patients (7%) had failure to thrive. EOS and CINF are found in 17% of children who had rectal biopsies negative for HSCR. Half of these children will need further medical input for the presence of persisting GI symptoms, food/milk allergy and failure to thrive, and the possibility to develop inflammatory bowel disease later in life.


The effect of mechanical stress on soft and hard tissue repair; a review.
The influence of mechanical forces on intact tissue is well established. A growing body of evidence demonstrates that healing wounds also respond to the functional demands of their mechanical environment. At the present time, an understanding of the fundamental mechanism by which mechanical stress affects tissues and wounds remains elusive.


Early administration of pyrrolidine dithiocarbamate extends the therapeutic time window of tissue plasminogen activator in a male rat model of embolic stroke.
Tissue plasminogen activator (tPA) is used in fewer than 4% of patients after ischemic stroke because of its narrow therapeutic time window. We tested whether pyrrolidine dithiocarbamate (PDTC), a drug with multiple mechanisms to provide neuroprotection, can be used to extend the therapeutic time window of tPA. Three-month-old male Sprague-Dawley rats were subjected to embolic stroke in the area supplied by the right middle cerebral artery. tPA at 10 mg/kg was given intravenously 4 h after the onset of stroke. PDTC at 50 mg/kg was given via gastric gavage at 30 min or 4 h after the onset of stroke. Two days after the stroke, neurological outcome was evaluated and the right frontal cortex area 1 (Fr1), an ischemic penumbral region, was harvested for analysis. PDTC given at 30 min after the stroke reduced infarct volumes and improved neurological functions no matter whether the rats received tPA. PDTC also reduced tPA-increased hemorrhagic volumes. Consistent with these results, PDTC in the presence or absence of tPA treatment attenuated the increase of proinflammatory cytokines, oxidative stress and matrix metalloprotease 2 activity in the right Fr1. However, PDTC given at 4 h after the onset of stroke did not improve the neurological outcome of rats treated with or without tPA. Our results suggest that PDTC given at an early time point but not in a delayed phase provides neuroprotection against embolic stroke and may be used to extend the therapeutic time window of tPA.


Characterization of CTLA4 Trafficking and Implications for Its Function.
CTLA4 is an essential negative regulator of T-cell immune responses and a key checkpoint regulating autoimmunity and antitumor responses. Genetic mutations resulting in quantitative defects in the CTLA4 pathway are also associated with the development of immune dysregulation syndromes in humans. It has been proposed that CTLA4 functions to remove its ligands CD80 and CD86 from opposing cells by a process known as transendocytosis. A quantitative characterization of CTLA4 synthesis, endocytosis, degradation, and recycling and how these affect its function is currently lacking. In a combined in vitro and in silico study, we developed a mathematical model and identified these trafficking parameters. Our model predicts optimal ligand removal in an intermediate affinity range. The intracellular CTLA4 pool as well as fast internalization, recovery of free CTLA4 from internalized complexes, and recycling is critical for sustained functionality. CD80-CTLA4 interactions are predicted to dominate over CD86-CTLA4. Implications of these findings in the context of control of antigen-presenting cells by regulatory T cells and of pathologic genetic deficiencies are discussed. The presented mathematical model can be reused in the community beyond these questions to better understand other trafficking receptors and study the impact of CTLA4 targeting drugs.


The evolution in the use of MIBG scintigraphy in pheochromocytomas and paragangliomas.
Radioiodinated metaiodobenzylguanidine (MIBG) was developed in the late 1970's, at the Michigan University Medical Center, for imaging of the adrenal medulla and its diseases. Soon after, MIBG was shown to depict a wide range of tumors of neural crest origin other than pheochromocytomas/paragangliomas (Pheo/PGL) with the result that its use rapidly spread to many countries. After more than 30 years of clinical application, MIBG continues to be the most widespread radiopharmaceutical for the functional imaging of Pheo/PGL in spite of the emergent role of PET agents for detection of these tumors. In this paper we review the evolution in the use of MIBG over more than 30 years of experimental and clinical applications, with particular focus on the uptake mechanisms, pharmacokinetics, biodistribution and drug interaction as well as on clinical studies in Pheo/PGL also in comparison to other gamma-emitters tracers and PET radiopharmaceuticals.


Using the NIH Research, Condition and Disease Categorization Database for research advocacy: Schizophrenia research at NIMH as an example.
In 2008 the National Institutes of Health established the Research, Condition and Disease Categorization Database (RCDC) that reports the amount spent by NIH institutes for each disease. Its goal is to allow the public "to know how the NIH spends their tax dollars," but it has been little used. The RCDC for 2018 was used to assess 428 schizophrenia-related research projects funded by the National Institute of Mental Health. Three senior psychiatrists independently rated each on its likelihood ("likely", "possible", "very unlikely") of improving the symptoms and/or quality of life for individuals with schizophrenia within 20 years. At least one reviewer rated 386 (90%), and all three reviewers rated 302 (71%), of the research projects as very unlikely to provide clinical improvement within 20 years. Reviewer agreement for the "very unlikely" category was good; for the "possible" category was intermediate; and for the "likely" category was poor. At least one reviewer rated 30 (7%) of the research projects as likely to provide clinical improvement within 20 years. The cost of the 30 projects was 5.5% of the total NIMH schizophrenia-related portfolio or 0.6% of the total NIMH budget. Study results confirm previous 2016 criticisms that the NIMH schizophrenia-related research portfolio disproportionately underfunds clinical research that might help people currently affected. Although the results are preliminary, since the RCDC database has not previously been used in this manner and because of the subjective nature of the assessment, the database would appear to be a useful tool for disease advocates who wish to ascertain how NIH spends its public funds.


Exploring Therapeutic Potential of Atorvastatin Against Gram-positive and Gram-negative Bacteria: In silico, In vitro and In vivo Evidences.
The incidences of opportunistic bacterial infections have increased in the past two decades or is expected to increase in the near future. Despite the availability of various classes of antibiotics, bacterial infections are not handled properly. Thus, in the present study, we have repurposed atorvastatin against various types of bacterial strains by using in silico, in vitro, and in vivo studies. Furthermore, a preliminary safety study was conducted using MTT assay. In silico study results have revealed that atorvastatin has good interaction with various targets of bacterial cell like that of the reference ligand. However, under in vitro conditions,it was found that atorvastatin was effective at higher concentrations (>128 μg/ml) against various bacterial strains. Thus, further, atorvastatin was tested in combination with standard antibiotics and has shown a synergistic effect. The MTT assay results have revealed non-cytotoxic activity of atorvastatin. In conclusion, atorvastatin in combination with standard drugs could be developed as an antibacterial agent.


Sequential or simultaneous visual processing deficit in developmental dyslexia?
The ability of dyslexic children with or without phonological problems to process simultaneous and sequential visual information was assessed using two tasks requiring the oral report of simultaneously or sequentially displayed letter-strings. The two groups were found to exhibit a simultaneous visual processing deficit but preserved serial processing skills. However, the impairment in simultaneous processing was larger in the dyslexic group with no phonological disorder. Although sequential and simultaneous processing skills both related to reading performance, simultaneous processing alone significantly contributed to reading speed and accuracy. These findings suggest that a simultaneous processing disorder might contribute to developmental dyslexia.


The competency quagmire: clarification of the nursing perspective concerning the issues of competence and informed consent.
Selected issues involved in informed consent and the patient's competence to give it are examined. A discussion of tests of competence reveals some of the problems encountered in using them. There are differences in medical and nursing functions and responsibilities in the informed consent process. One nursing framework, self care, is used to discuss whether informed consent should be obtained from patients when they receive specific nursing care.


Impact of initial fluid resuscitation volume on clinical outcomes in patients with heart failure and septic shock.
Fluid resuscitation is a key treatment for sepsis, but limited data exists in patients with existing heart failure (HF) and septic shock. The objective of this study was to determine the impact of initial fluid resuscitation volume on outcomes in HF patients with reduced or mildly reduced left ventricular ejection fraction (LVEF) with septic shock. This multicenter, retrospective, cohort study included patients with known HF (LVEF ≤50%) presenting with septic shock. Patients were divided into two groups based on the volume of fluid resuscitation in the first 6 h; <30 mL/kg or ≥30 mL/kg. The primary outcome was a composite of in-hospital mortality or renal replacement therapy (RRT) within 7 days. Secondary outcomes included acute kidney injury (AKI), initiation of mechanical ventilation, and length of stay (LOS). All related data were collected and compared between the two groups. A generalized logistic mixed model was used to assess the association between fluid groups and the primary outcome while adjusting for baseline LVEF, Acute Physiology and Chronic Health Evaluation (APACHE) II score, inappropriate empiric antibiotics, and receipt of corticosteroids. One hundred and fifty-four patients were included (93 patients in <30 mL/kg group and 61 patients in ≥30 mL/kg group). The median weight-based volume in the first 6 h was 17.7 (12.2-23.0) mL/kg in the <30 mL/kg group vs. 40.5 (34.2-53.1) mL/kg in the ≥30 mL/kg group (P <0.01). No statistical difference was detected in the composite of in-hospital mortality or RRT between the <30 mL/kg group compared to the ≥30 mL/kg group (55.9% vs. 45.9%, P=0.25), respectively. The <30 mL/kg group had a higher incidence of AKI, mechanical ventilation, and longer hospital LOS. In patients with known reduced or mildly reduced LVEF presenting with septic shock, no difference was detected for in-hospital mortality or RRT in patients who received ≥30 mL/kg of resuscitation fluid compared to less fluid, although this study was underpowered to detect a difference. Importantly, ≥30 mL/kg fluid did not result in a higher need for mechanical ventilation.


Ceruloplasmin gene expression in human cancer cells.
The copper transport protein, ceruloplasmin, is suggested to have a role in cancer since it is involved in angiogenesis and neovascularization. In order to understand the role of ceruloplasmin in malignant cells, we have recently isolated and sequenced a human ceruloplasmin cDNA clone. In the present study, we have investigated the ceruloplasmin gene expression in human colon and breast cancer cell lines. The poly (A) RNA from human colon (WiDr) and human breast (MCF-7) cancer cell lines was analyzed for the presence of ceruloplasmin mRNA. The Northern blot analysis revealed the presence of a 3.7 kb band of ceruloplasmin mRNA in these cell lines. Dot blot analysis revealed that ceruloplasmin mRNA is at least three fold more abundant in tumor cells as compared to normal rat liver.


MAOA-uVNTR genotype predicts interindividual differences in experimental aggressiveness as a function of the degree of provocation.
The MAOA-uVNTR has been suggested to play a role regarding aggression, however, results are inconsistent. We aimed at further elucidating potential effects of the MAOA-uVNTR on aggressiveness with respect to potential modulators: sex, experimental vs. trait aggressiveness and type of aggressiveness (proactive vs. reactive aggressiveness). We tested 239 healthy young adults (88 men/151 women). Participants were genotyped for the MAOA-uVNTR and performed a modified version of a competitive reaction time task - a commonly used and well established tool to elicit and measure aggressiveness. Furthermore, they completed a self-report scale measuring trait aggressiveness. We found a main effect of MAOA-uVNTR on a measure of reactive aggressiveness for both men and women, whereby the low-activity alleles of the MAOA-uVNTR were associated with substantially increased aggressive reactions (p<.05). This effect was unique for reactive aggressiveness. Measures of proactive aggressiveness or self reports were not associated with the MAOA-uVNTR-genotype. Our data are in line with earlier studies and indicate the MAOA-uVNTR-genotype to be specifically associated with measures of reactive impulsive experimental aggressiveness in healthy men and women. Furthermore the association between the MAOA-uVNTR genotype and aggressive responses increases in a fashion linear to the degree of provocation. This indicates that the low-functional alleles of the MAOA-uVNTR are not associated with increased aggressive behavior per se, but rather with an increased aggressive reactivity to provocation.


Expanding application of CRISPR-Cas9 system in microorganisms.
The development of CRISPR-Cas9 based genetic manipulation tools represents a huge breakthrough in life sciences and has been stimulating research on metabolic engineering, synthetic biology, and systems biology. The CRISPR-Cas9 and its derivative tools are one of the best choices for precise genome editing, multiplexed genome editing, and reversible gene expression control in microorganisms. However, challenges remain for applying CRISPR-Cas9 in novel microorganisms, especially those industrial microorganism hosts that are intractable using traditional genetic manipulation tools. How to further extend CRISPR-Cas9 to these microorganisms is being an urgent matter. In this review, we first introduce the mechanism and application of CRISPR-Cas9, then discuss how to optimize CRISPR-Cas9 as genome editing tools, including but not limited to how to reduce off-target effects and Cas9 related toxicity, and how to increase on-target efficiency by optimizing crRNA and sgRNA design.


The interdependent transport of yeast vacuole Ca2+ and H+ and the role of phosphatidylinositol 3,5-bisphosphate.
Yeast vacuoles are acidified by the v-type H+-ATPase (V-ATPase) that is comprised of the membrane embedded VO complex and the soluble cytoplasmic V1 complex. The assembly of the V1-VO holoenzyme on the vacuole is stabilized in part through interactions between the VO a-subunit ortholog Vph1 and the lipid phosphatidylinositol 3,5-bisphosphate (PI(3,5)P2). PI(3,5)P2 also affects vacuolar Ca2+ release through the channel Yvc1 and uptake through the Ca2+ pump Pmc1. Here, we asked if H+ and Ca2+ transport activities were connected through PI(3,5)P2. We found that overproduction of PI(3,5)P2 by the hyperactive fab1T2250A mutant augmented vacuole acidification, whereas the kinase-inactive fab1EEE mutant attenuated the formation of a H+ gradient. Separately, we tested the effects of excess Ca2+ on vacuole acidification. Adding micromolar Ca2+ blocked vacuole acidification, whereas chelating Ca2+ accelerated acidification. The effect of adding Ca2+ on acidification was eliminated when the Ca2+/H+ antiporter Vcx1 was absent, indicating that the vacuolar H+ gradient can collapse during Ca2+ stress through Vcx1 activity. This, however, was independent of PI(3,5)P2, suggesting that PI(3,5)P2 plays a role in submicromolar Ca2+ flux but not under Ca2+ shock. To see if the link between Ca2+ and H+ transport was bidirectional, we examined Ca2+ transport when vacuole acidification was inhibited. We found that Ca2+ transport was inhibited by halting V-ATPase activity with Bafilomycin or neutralizing vacuolar pH with chloroquine. Together, these data show that Ca2+ transport and V-ATPase efficacy are connected but not necessarily through PI(3,5)P2.


Transurethral reconstruction of fossa navicularis strictures with dorsal inlay buccal mucosa graft urethroplasty.
To describe a single stage, glans-sparing urethroplasty technique for fossa navicularis strictures using a transurethral dorsal inlay buccal mucosa graft. We conducted a retrospective review of a prospectively maintained urethral stricture database to identify all fossa navicularis strictures reconstructed with a single stage, transurethral dorsal inlay buccal mucosa graft urethroplasty (5/2015-6/2020). Primary outcomes were anatomic success, defined as the ability to pass a 17 Fr flexible cystoscope, and functional success, defined as the lack of obstructive voiding symptoms and no need for further procedures. Secondary outcomes were postoperative complications and patient satisfaction. Sixteen patients were included. Mean age was 63.1 years (43.9-75.6) and mean stricture length was 1.7 cm (1.4-2.0). Stricture etiology included internal trauma (62.5%), idiopathic (25.0%), and lichen sclerosus (12.5%). Prior endoscopic procedures were done in 75% of patients. Over a median follow-up of 28.8 months (IQR 17.6-38.0), anatomic success was 15/16 (93.8%) and functional success was 16/16 (100%). The single anatomic recurrence was at 4.2 months postoperatively. No additional procedures were required. Urinary tract infection occurred in 25% (4/16). There were no instances of de novo erectile dysfunction, chordee, or wound infection. All patients would recommend urethroplasty to others and all patients were either very satisfied (83.3%) or satisfied (16.7%) with the procedure. Penile sensitivity was unchanged in 83.3%, increased in 8.3% and decreased in 8.3%. Transurethral dorsal inlay buccal mucosa graft urethroplasty is a viable option for reconstruction of fossa navicularis strictures that avoids splitting the glans and results in excellent cosmesis.


Morphological Characterization and Molecular Phylogenetic Analysis of Kudoa iwatai from Large Yellow Croaker (Larimichthys crocea) as a New Host in China.
Kudoa (Myxosporea: Multivalvulida) parasites are critical pathogens in marine and freshwater fish associated with significant economic losses and reduced market prices caused by post-mortem myoliquefaction or numerous cysts on muscles. In the present study, large yellow croakers infected by Kudoa were found during fish disease surveillance in China in November 2020 and used for morphological observation and characterization using light DIC microscopy and electron microscopy. Numerous creamy-white oval plasmodia were observed in muscles and on the surface of brain cartilage, gill arches, and serosal surfaces. The spores were considerably longer and thicker than previously reported Kudoa, with protruding polar filaments (PFs) in the mature spores, fingertip-shaped apical projections (APs), and polar capsules. Phylogenetic analyses with SSU rDNA, LSU rDNA, and mitochondrial DNA showed that the Kudoa-infected sample (LcK-2020) had the highest similarity to Kudoa iwatai reported in Japan. Based on the morphological characterization and phylogenetic analysis, it could be concluded that the sample LcK-2020 was infected by Kudoa iwatai, which would be the first report of Kudoa iwatai infection in large yellow croaker in China.


The roles of K+-dependent Na+/Ca2+ exchanger 2 (NCKX2) in methamphetamine-induced behavioral sensitization and conditioned place preference in mice.
Drug addiction, including methamphetamine (METH) addiction, is a significant public health and social issue. Perturbations in intracellular Ca2+ homeostasis are associated with drug addiction. K+-dependent Na+/Ca2+ exchanger 2 (NCKX2) is located on neuronal cell membranes and constitutes a Ca2+ clearance mechanism, with key roles in synaptic plasticity. NCKX2 is associated with motor learning, memory, and cognitive functions. However, the role of NCKX2 in METH addiction remains unclear. In this study, we investigated the expression levels of NCKX2 in four addiction-related brain regions: the prefrontal cortex (PFc), nucleus accumbens (NAc), dorsal striatum (DS), and hippocampus (Hip) in a C57/BL6 mouse model of METH-induced conditioned place preference (CPP) and behavioral sensitization. Levels of NCKX2 were unchanged in these brain regions in mice with METH-induced CPP but were decreased in the PFc and NAc of mice with METH-induced behavioral sensitization. Adeno-associated virus (AAV)-mediated overexpression of NCKX2 in the PFc attenuated the expression phase of METH-induced behavioral sensitization in mice, whereas AAV-mediated knockdown of NCKX2 enhanced the effects of METH. Collectively, our results suggest that NCKX2 is involved in METH-induced behavioral sensitization but does not affect conditioned reward-related memory, highlighting the potential of NCKX2 as a molecular target for studying the mechanisms underscoring METH addiction.


Two waves of antigen-containing dendritic cells in vivo in experimental Leishmania major infection.
Dendritic cells (DC) can induce Th1 cell differentiation by producing IL-12. In experimental infection with Leishmania major, DC could differently respond to infection and induce Th1 cells in C57BL/6 but not BALB/c mice, and thus determine the resistance or susceptibility of these mice. We characterized L. major antigen-containing DC in vivo in draining lymph nodes of both strains. Conventional experimental infection is shown to result in two waves of these DC and our data argue against a relevant genetic difference in the DC initiating the anti-parasite Th cell response in these mice. In both strains the first wave of DC presented L. major antigens but was not infected, produced IL-12 but induced disease-mediating Th2 cells upon adoptive transfer. In contrast to current belief, this response was therefore not initiated by infected DC, which were only detected in the second wave. The kinetics of the two waves suggests that DC turnover has an important impact on antigen presentation during infections with complex pathogens.


Enantiomeric determination of pantoprazole in human plasma by multidimensional high-performance liquid chromatography.
Multidimensional HPLC is a powerful tool for the analysis of samples of a high degree of complexity. This work reports the use of multidimensional HPLC by coupling a RAM column with a chiral polysaccharide column to the analysis of Pantoprazole in human plasma by direct injection. The enantiomers from the plasma samples were separated with high resolution on a tris(3,5-dimethoxyphenylcarbamate) of amylose phase after clean-up by a RAM BSA octyl column. Water was used as solvent for the first 5 min in a flow-rate of 1.0 ml/min for the elution of the plasmatic proteins and then acetonitrile-water (35:65 v/v) for the transfer and analysis of pantoprazole enantiomers, which were detected by UV at 285 nm. Analysis time was 28 min with no time spent on sample preparation. A good linear relationship was obtained in the concentration range of 0.20 to 1.5 microg/ml for each enantiomer. Inter and intra-day precision and accuracy were determined by one low (0.24 microg/ml), one medium (0.70 microg/ml) and one high (1.3 microg/ml) plasma concentration and gave a C.V. varying from 1.80 to 8.43% and accuracy from 86 to 92%. Recoveries of pantoprazole enantiomers were in the range of 93.7-101.2%. The validated method was applied to the analysis of the plasma samples obtained from ten Brazilian volunteers who received an 80 mg oral dose of racemic pantoprazole and was able to quantify the enantiomers of pantoprazole in all clinical samples analyzed.


Processing fluency affects subjective claims of recollection.
Previous studies that have used the remember-know paradigm to investigate subjective awareness in memory have shown that fluency manipulations have an impact on "know" responses but not on "remember" responses (e.g., Rajaram, 1993), a finding typically accounted for by invoking inferential processing in judgments of familiarity but not of recollection. However, in light of several researchers' criticisms of this procedure, as well as findings documenting the influence of processing fluency on various subjective judgments, the present study was conducted in order to investigate whether judgments of recollection might also be subject to inferential processes and not solely the product of conscious retrieval. When the standard remember-know procedure was used (Experiment 1), manipulations of perceptual fluency increased "know" responses but had no effect on "remember" responses, replicating previous findings. However, when an independent ratings method was employed (Higham & Vokey, 2004), manipulations of perceptual fluency (Experiment 2) and conceptual fluency (Experiment 3) reliably increased claims of both familiarity and recollection, suggesting that the conclusion that fluency affects only "know" responses may be an artifact of the standard remember-know procedure.


Variation of LEF-1 mRNA expression in low-grade B-cell non-Hodgkin's lymphoma.
During normal B-lymphocyte development once cells pass the pro-B stage the transcription factor LEF-1, a key component of the Wnt/beta-catenin pathway, is down regulated. However studies have shown that B-cell chronic lymphocytic leukaemia (CLL) lymphocytes, which have a mature B-cell phenotype, still express abundant LEF-1. This study demonstrates that although LEF-1 mRNA is universally, highly expressed in B-CLL, expression of this gene is much lower or absent in the majority of low-grade B-cell non-Hodgkin's lymphoma (NHL). This suggests that there exist key differences in the activity of the Wnt/beta-catenin pathway between low-grade B-cell malignancies.


[Peri-ano-rectal cysts and tumours of vestigial origin in the adult (author's transl)].
Four cases of cysts or tumours developing from embryological remnants in the peri-ano-rectal space are reported. The authors propose for these rare lesions a coherent and simplified classification in two main groups: teratomas on the one hand, developmental cysts on the other hand. The various methods of diagnosis are studied. The precise diagnosis of a congenital cyst can only be made by pathological examination of the specimen after surgical removable. Any rapid local change implied malignant degeneration. Such a risk suggests immediate removal of any known ano-rectal cyst or tumour. The route of approach is usually perineal. Kraske's position gives good access to the retro-ano-rectal space. Possible pelvic spread of these lesions sometimes requires the use of the abdominal approach, either straight away or secondarily when total removal is impossible by the perineal route alone.


Amino acid metabolism, transport and signalling in the liver revisited.
The liver controls the systemic exposure of amino acids entering via the gastro-intestinal tract. For most amino acids except branched chain amino acids, hepatic uptake is very efficient. This implies that the liver orchestrates amino acid metabolism and also controls systemic amino acid exposure. Although many amino acid transporters have been identified, cloned and investigated with respect to substrate specificity, transport mechanism, and zonal distribution, which of these players are involved in hepatocellular amino acid transport remains unclear. Here, we aim to provide a review of current insight into the molecular machinery of hepatic amino acid transport. Furthermore, we place this information in a comprehensive overview of amino acid transport, signalling and metabolism.


Behavioral effects of a LHRH antagonist in intact and ovariectomized rats.
The effects of the LHRH antagonism on the acquisition of conditioned responses (CARs) and spontaneous motility were studied in intact and ovariectomized rats. A synthetic antagonist of LHRH, [N-acetyl-D-p-chloro-Phe1,2,D-Trp3,D-Arg6,D-Ala10]-LHRH, was injected in a single dose (10 micrograms/rat, SC) at noon on the day of proestrus in the normally cycling rat, and behavioral experiments were carried out on the morning of estrus or metestrus. Two procedures were followed in the ovariectomized rats: in the first, the antagonist was injected 1 h before estradiol, and in the second, at noon on the day after estradiol replacement. The experiments were carried out 24 and 48 h after estradiol, respectively. The LHRH antagonist facilitated the acquisition of CARs in both experimental groups, thus reversing the impairments observed during estrus and metestrus and those induced by estradiol replacement. The antagonist decreased the number of head shakes during estrus, whereas it induced an increase in total motility and rears in ovariectomized control animals. On the other hand, the antagonist increased the number of rears and reversed the decrease in grooming behavior induced by estradiol. The results led to the idea of a role of LHRH in behaviors not apparently related to sex, which could explain the behavioral changes observed across the estrous cycle and those induced by estradiol replacement in ovariectomized rats.


Evidence for avian influenza A infections among Iowa's agricultural workers.
Identifying risk factors for zoonotic influenza transmission may aid public health officials in pandemic influenza planning. We sought to evaluate rural Iowan agriculture workers exposed to poultry for previous evidence of avian influenza virus infection. In 2004 we enrolled 803 rural adult Iowans in a 2-year prospective study of zoonotic influenza transmission. Their enrollment data and sera were compared with those from 66 adult controls enrolled at the University of Iowa in 2006 by proportional odds modeling. The 803 participants were 58.8% male with a mean age of 55.6 yrs. Forty-eight percent reported previous poultry exposure. Sera were studied by microneutralization techniques for antibodies against avian H4, H5, H6, H7, and H9 viruses. Touching live birds was associated (OR = 1.2; 95% CI 1.02-1.8) with increased antibody titer against H5 virus. Similarly, participants who reported hunting wild birds had increased antibody titers against H7 virus (OR = 2.8; 95%CI = 1.2-6.5) and subjects who reported recent work with poultry had increased antibody titers against H6 (OR = 3.4; 95% CI 1.4-8.5) and H7 viruses (OR = 2.5, 95% CI = 1.1-5.7). There was no evidence of elevated antibody against avian H4 or H9 viruses. These data suggest that hunting and exposure to poultry may be important risk factors for avian influenza virus infection among rural US populations. Agriculture workers should be included in influenza pandemic plans.


Corrigendum: QUaRTM: A Quadcopter with Unactuated Rotor Tilting Mechanism capable of faster, more agile, and more efficient flight.
[This corrects the article DOI: 10.3389/frobt.2022.1033715.].


Attitudes among medical and nonmedical university students toward the survivors of childhood abuse.
Childhood sexual abuse (CSA) is increasing, and current studies should address issues such as stigmas and casual attitudes that may be amenable to intervention. Thus, we aimed to investigate the effects of education on the attitudes of medical students toward CSA survivors. We divided medical students into 2 groups: preclinical and clinical medical students. We compare the attitudes of nonmedical students with the attitudes of medical students with respect to survivors of CSA. All participants were asked to complete a questionnaire about stigmatization. There were 572 students in our study, 347 of whom were enrolled in medical school. Of the 347 medical students, 151 were from the preclinical group, and 196 were from the clinical practice group. The remainder of the participants (n = 225) were in other fields of university. Of the medical students, 68.3% responded negatively about "asking a survivor of abuse to supervise their child for a few hours," whereas 62.7% of the nonmedical students responded negatively. When asked, "What would you think if your child wanted to marry a survivor of sexual abuse?", 72.9% of the medical students and 59.1% of the nonmedical students displayed negative attitudes toward that idea. Our research also found that significantly more preclinical students than clinical students would not want to be in a companionship/friendship with a CSA survivor at work (P = .015) or in their neighborhoods (P = .034). Both medical and nonmedical students seem to have negative thoughts about sexual abuse survivors, particularly when the issue involves their own children and/or social situations. Thus, the medical professionals must be careful and should avoid over interpretations of findings on childhood abuse studies that may lead to stigmatization.


Lipidomic changes in mouse oocytes vitrified in PEG 8000-supplemented vitrification solutions.
Cryopreserved oocytes are inevitably exposed to cold stress, which negatively affects the cellular aspects of the oocytes. Lipidomic analysis of the oocytes reveals quantitative changes in lipid classes under conditions of cold stress, leading to potential freezing-vulnerability. We had previously shown that specific phospholipids are significantly downregulated in vitrified-warmed mouse oocytes compared to those in fresh oocytes. In this study, we examined whether supplementation of polyethylene glycol 8000 (PEG 8000) during vitrification influences the lipidome of the oocytes. We used liquid chromatography with tandem mass spectrometry (LC-MS/MS) to study the alteration in the lipidome in three groups of mouse oocytes: fresh, vitrified-warmed, and vitrified with PEG 8000-warmed during vitrification. In these groups, we targeted to analyze 21 lipid classes. We profiled 132 lipid species in the oocytes and statistical analyses revealed lipid classes that were up- or downregulated in these groups. Overall, our data revealed that several classes of lipids were affected during vitrification, and that oocytes vitrified with PEG 8000 to some extent alleviated the levels of changes in phospholipid and sphingolipid contents during vitrification. These results suggest that phospholipids and sphingolipids are influenced by PEG 8000 during vitrification and that PEG 8000 can be considered as a potential candidate for preserving membrane integrity during oocyte cryopreservation.


Reversible alopecia after single-dose methotrexate treatment in a patient with ectopic pregnancy.
To report a case of a women who received a single 50 mg/m2 dose of methotrexate (MTX) on two occasions for two separate ectopic pregnancies. After both treatments the patient experienced reversible alopecia. Case report. Single human patient in a university-run clinic. A patient with an ectopic pregnancy presenting for treatment on two separate occasions. A single dose of 50 mg/m2 MTX given to the patient on both occasions. Loss of scalp hair. On two separate occasions the patient lost a significant portion (33% to 50%) of her scalp hair. Even single-dose MTX as used for the treatment of ectopic pregnancy may induce alopecia in a susceptible patient.


[Mitochondrial cardiomyopathy with a high degree of heart muscle hypertrophy].
Because of cardiomyopathy with concomitant cardiac hypertrophy, a 21 month-old girl died 7 days following operative closure of a cleft palate. The cardiomyopathy was due to an excessive increase of enlarged and abnormally structured mitochondria. Similar mitochondrial change were found in the skeletal muscles (Mitochondrial Myopathy).


The effect of patient compliance on remission rates in pediatric noninfectious uveitis.
To identify barriers to compliance in pediatric noninfectious uveitis, and to examine its association with achieving steroid-free remission. A retrospective analysis was performed on pediatric patients with noninfectious uveitis on immunomodulatory therapy treated at the University of Texas Southwestern Medical School and Children's Medical Center (Dallas, TX) between September 2015 and March 2017. Compliance barriers were identified and rates of achieving steroid-free remission were calculated. A total of 57 patients with noninfectious uveitis requiring immunosuppressive therapy met inclusion criteria. Thirty-three (58%) of patients were compliant. Notable barriers to compliance included regimens requiring >3 medications, patient/parent negligence, transportation issues, family strife, and presence of an associated systemic autoimmune disease (P < 0.050). At a median follow-up of 24 months, a total of 28 (49%) achieved steroid-free remission. The presence of 3 or more compliance barriers was associated with decreased remission rates (P < 0.050). Poor compliance was associated with decreased rates of steroid-free remission (21% vs 79% [P = 0.002]). Noncompliant patients with noninfectious pediatric uveitis requiring immunomodulatory therapy were found to have a lower rate of achieving steroid-free remission compared to patients who exhibited full compliance.


Impact of sleep-disordered breathing on life-threatening ventricular arrhythmia in heart failure patients with implantable cardioverter-defibrillator.
It was recognized that sleep-disordered breathing (SDB) is associated with cardiac arrhythmia and sudden cardiac death. However, it was unclear whether the presence of SDB increased the risk of life-threatening ventricular arrhythmia in patients with heart failure (HF) with an implantable cardioverter-defibrillator (ICD). Seventy-one patients with HF and an ICD who were followed up for 180 days after a sleep study were prospectively studied. All patients had an ejection fraction <or=35%. SDB was defined as an apnea-hypopnea index >or=10 events/hour on the sleep study. The frequency of appropriate ICD therapy and the day-night pattern of ICD therapies were compared between patients with and without SDB. SDB was diagnosed in 47 of 71 patients (66%). There were no statistical differences between patients with and without SDB in baseline cardiac function. However, appropriate ICD therapies occurred more frequently in patients with (43%) than without SDB (17%; p = 0.029). On multivariate analysis, the presence of SDB was an independent predictor for appropriate ICD therapy (hazard ratio 4.05, 95% confidence interval 1.20 to 13.65, p = 0.015). Moreover, the rate of total ICD therapy from midnight to 6 A.M. was significantly higher in patients with (34%) than without SDB (13%; p = 0.046). In conclusion, in patients with HF with an ICD, the presence of SDB was common and an independent predictor of life-threatening ventricular arrhythmias that were more likely to occur during sleep.


AHA surveys major source of hospital data.
The Annual Survey of Hospitals and the National Hospital Panel Survey work together to provide a thorough picture of hospital activities.


Does cigarette smoking affect the diagnostic reliability of hemoglobin alpha 2 delta 2 (HbA2)?
Quantitation of hemoglobin alpha 2 delta 2 (HbA2) is a basic and confirmatory test in diagnosing the carrier state of beta-thalassemia. The present study was designed to investigate the effect of cigarette smoking on the diagnostic reliability of HbA2. A total of 2,867 (654 smokers and 2,213 never smokers) male subjects were involved in the present study. The subjects were categorized into three groups according to their laboratory findings: beta-thalassemia minor, iron deficient, and normal groups. Complete blood count (CBC) parameters and HbA2 levels were compared between smokers and never smokers of each group according to the independent-samples t-test using the SPSS program, significance results were reported at P<0.05. The results showed a significant increase in red blood cell (RBC) mass (RBC count and hematocrit [Hct]) and Hb concentration in smokers of all groups; however, no significant differences were reported in the HbA2 level between smokers and never smokers in all groups. It was concluded that cigarette smoking does not affect the diagnostic reliability of the HbA2 test.


A cut-off value for the operation time and other risk factors in terms of the infection risk for flexible ureterorenoscopy.
To investigate the operation time (OT) and ureteral access sheath (UAS) usage with the infection rates and to determine a cut-off value for OT. We retrospectively analysed the data of the patients who underwent flexible ureterorenoscopy (FURS) for renal stones larger than 20 mm between 2010 and 2019. The investigated parameters were OT, UAS using, and infection status. The data were analysed by forming two groups according to whether the OT was less than 60 minutes and more, whether the UAS was used and whether an infection occurred. In addition, independent risk factors that may affect postoperative urinary infection development were also investigated by logistic regression analysis. And, a Receiver Operating Characteristic (ROC) curve analysis was applied to determine a cut-off value in OT terms, where infection rates increase more. A total of 575 patients were enrolled in the study. The rates of the usage of UAS and infection were greater statistically in the group for longer than 60 minutes. OT was longer statistically in the infection group than in the group without infection (94.1 ± 14.2 and 68.01 ± 23.1, for groups 1 and 2, respectively, P < .05, Table 2). OT was statistically longer in the UAS group than the unused one (79.3 ± 24.4 and 66.7 ± 22.4, for groups 1 and 2, respectively, P < .05, Table 3). ROC analyses revealed a cut-off point of 87.5 min for OT in terms of infection rate. While the infection risk increases when OT exceeds 60 minutes, FURS can be safely performed up to 87.5 minutes with 89% sensitivity and 69% specificity infection risk.


Sensitivity-enhanced 2D IPAP, TROSY-anti-TROSY, and E.COSY experiments: alternatives for measuring dipolar 15N-1HN couplings.
Sensitivity-enhanced versions of the IPAP, TROSY-anti-TROSY, and E.COSY experiments for measuring one-bond 15N-1HN couplings are presented. Together with the previously developed sensitivity-enhanced E.COSY-type HSQC experiment they comprise a suite of sensitivity-enhanced experiments that allows one to chose the optimal spectrum for accurate measurement of one-bond 15N-1HN residual dipolar couplings in proteins. Since one-bond 15N-1HN residual dipolar couplings play uniquely important roles in structural NMR, these additional methods provide further tools for improving structure determination of proteins and other biological macromolecules.


Epidermal Langerhans cells in small fiber neuropathies.
We quantified the immune histiocytic Langerhans cells (LCs) in skin biopsy samples of patients with distal small fiber neuropathy (SFN). Patients were divided according to the presence or absence of neuropathic pain (burning pain) assessed by a visual analogue scale (VAS). We studied 13 diabetic patients (pain-DSFN), 7 nondiabetic patients (pain-SFN) who reported relevant neuropathic pain (VAS ≥ 3), and 6 nondiabetic patients without neuropathic pain (no-pain-SFN). Using double immunofluorohistochemistry with the PGP 9.5 and the langerin/CD207, we quantified the intraepidermal nerve fibers density (IENFD) and LCs per square millimeter in the epidermis. A group of 10 skin samples from healthy subjects served as controls. Confocal analysis was performed to evaluate LC PGP 9.5-immunoreactivity. We found a mean value of 334.3LC/mm(2) in controls, 310.2LC/mm(2) in no-pain-SFN, 329.6LC/mm(2) in pain-SFN and 484.3LC/mm(2) in pain-DSFN (analysis of variance; P=.01). In patients, analysis of covariance adjusted by different covariables showed that the presence of diabetes (F=5.2, P=.03) was associated with an increased number of LC/mm(2). There was a negative correlation between the IENFD and the number of LCs (r(2)=-0.13, P=.03). No statistically significant differences were found among groups of subjects either for the co-localization or for the number of LCs that were PGP 9.5-immunoreactive (analysis of variance; P>.05). These results indicate that patients with neuropathic pain in the context of SFN, specially those who had diabetes (DSFN), had an increased number of LCs in the epidermis that may play a role in the generation or maintenance of neuropathic pain.


Retrograde transport by the microtubule-associated protein MAP 1C.
Microtubules are involved in several forms of intracellular motility, including mitosis and organelle movement. Fast axonal transport is a highly ordered form of organelle motility that operates in both the anterograde (outwards from the cell body) and retrograde (from the periphery towards the cell body) direction. Similar microtubule-associated movement is observed in non-neuronal cells, and might be involved in secretion, endocytosis and the positioning of organelles within the cell. Kinesin is a mechanochemical protein that produces force along microtubules in an anterograde direction. We recently found that the brain microtubule-associated protein MAP 1C (ref. 7) is a microtubule-activated ATPase and, like kinesin, can translocate microtubules in an in vitro assay for microtubule-associated motility. MAP 1C seemed to be related to the ciliary and flagellar ATPase, dynein, which is thought to produce force in a direction opposite to that observed for kinesin. Here we report that MAP 1C, in fact, acts in a direction opposite to kinesin, and has the properties of a retrograde translocator.


Metabolic profiles of lung adenocarcinoma via peripheral blood and diagnostic model construction.
The metabolic profile of cancerous cells is shifted to meet the cellular demand required for proliferation and growth. Here we show the features of cancer metabolic profiles using peripheral blood of healthy control subjects (n = 78) and lung adenocarcinoma (LUAD) patients (n = 64). Among 121 detected metabolites, diagnosis of LUAD is based on arginine, lysophosphatidylcholine-acyl (Lyso.PC.a) C16:0, and PC-diacyl (PC.aa) C38:3. Network analysis revealed that network heterogeneity, diameter, and shortest path were decreased in LUAD. On the contrary, these parameters were increased in advanced-stage compared to early-stage LUAD. Clustering coefficient, network density, and average degree were increased in LUAD compared to the healthy control, whereas these topologic parameters were decreased in advanced-stage compared to early-stage LUAD. Public LUAD data verified that the genes encoding enzymes for arginine (NOS, ARG, AZIN) and for Lyso.PC and PC (CHK, PCYT, LPCAT) were related with overall survival. Further studies are required to verify these results with larger samples and other histologic types of lung cancer.


Exploring the links among borderline personality disorder symptoms, trauma, and pain in patients with chronic pain disorders.
Chronic pain and borderline personality disorder (BPD) are commonly comorbid and jointly associated with increased symptoms of both disorders and clinical and functional impairment. Little is known, however, about specific links between these disorders. In a cross-sectional study of patients with chronic pain, we compared participants high or low on BPD symptoms on patterns of pain experience and types of child and adult traumas. Adults (N = 181) with chronic pain completed self-reports of pain severity, dimensions of pain experiencing, body coverage of pain, and clinical indicators of central sensitization (i.e., chronic hypersensitivity of the central nervous system), as well as measures of child and adult physical abuse, sexual abuse, trauma, and neglect. Participants also completed the McLean Screening Instrument for BPD. Participants with clinically significant BPD symptoms (n = 32) reported more childhood sexual trauma, punishment, and neglect, as well as adult physical/sexual trauma, than those without elevated BPD symptoms. Among participants with clinically significant BPD symptoms, affective pain and central sensitization were elevated, potentially explained by heightened negative affect in BPD. BPD symptoms are associated with increased clinical severity among patients with chronic pain as well as a unique manifestation of pain experiencing (i.e., increased affective pain and central sensitization in particular). Childhood trauma of all types is associated with chronic pain and BPD co-occurrence. Researchers and clinicians should assess for BPD in people with chronic pain to enhance conceptual models of the transaction between these disorders and to improve clinical care.


Solitary hot spots in the ribs on bone scan: value of thin-section reformatted computed tomography to exclude radiography-negative fractures.
The purpose of this study was to classify solitary, scintigraphy-positive and radiography-negative rib lesions and to clarify the features of rib fractures by using thin-section reformatted helical computed tomography (CT). Forty-seven patients whose whole-body bone scans showed a solitary hot spot in a rib as their first abnormal scintigraphic finding and whose plain radiograph showed negative or indeterminate results were examined by thin-section reformatted CT. Final diagnosis was based on follow-up over a 20-month period. The final diagnosis included 17 cases of fractures where CT findings were fracture line, focal sclerosis, and callus formation. Fourteen ribs demonstrated intramedullary, focal osteosclerosis, and 8 ribs did not demonstrate any abnormalities. Four metastatic lesions appeared as intramedullary mixture of osteolysis and osteosclerosis, or bone destruction. Four intramedullary lesions with cystic appearance remained unchanged. Thin-section reformatted helical CT delineated minute, radiographically occult fractures of the rib. Excluding fractures, helical CT can avoid further unnecessary examinations.


Sonographic characterization of 271 hepatic hemangiomas with typical appearance on CT imaging.
Hepatic hemangioma is the most frequent benign solid tumor that requires differentiation from hepatic malignancy on ultrasonography. Useful ultrasound findings are therefore required for diagnosis. The following factors were investigated for 271 masses diagnosed as hepatic hemangioma by contrast-enhanced computed tomography (CT) in 188 patients: hepatic subsegment location, shape, maximum diameter, internal echo level, hyperechoic rim, posterior echoes, marginal hypoechoic band (halo), lateral shadow, blood flow signal in the central portion of the mass, and underlying liver disease. Hepatic masses were classified by internal echo pattern as homogeneous hyperechoic (35.0%), homogeneous hypoechoic (22.9%), isoechoic (5.2%), mixed hyperechoic (22.1%), or mixed hypoechoic (14.8%) masses. Twelve masses were isoechoic, and could be recognized by the hyperechoic rim. Posterior echo enhancement was present in 66 masses. There were no cases of posterior echo attenuation. A halo was present in six masses, with the underlying condition being fatty infiltrated liver in two masses; a hypoechoic region with a small amount of fat deposit was present around the mass, and this resembled a halo. This finding was termed a "pseudohalo." There were no masses in which a lateral shadow was observed. In terms of blood flow signals in the central portion of the mass, pulsatile flow was present in only one mass. Important ultrasonographic findings of hepatic hemangioma are characterized as the absence of lateral shadow (100%) and no attenuation of posterior echoes (100%), while the presence of a hyperechoic rim is useful for detecting isoechoic hemangioma.


Role of plants in metal cycling in a tidal wetland: implications for phytoremidiation.
Accumulation of 8 metals and the semimetal As in 29 plant species was quantified in a restored tidal wetland on a contaminated site. Transfer coefficients between sediment and aboveground plant tissues were lower than in many other systems; from 0.013 (Pb) to 0.189 (Mn). A minor fraction of the sediment metal pool cycled through the aboveground vegetation (≤0.02%). However, during the four years of this study, species composition changed, and plant biomass as well as the metal pool in the vegetation increased (≤0.12%). Succession to either a willow dominated brushwood or a monospecific reed stand can further enlarge this pool (2.5%). Since the amount of trace metals in the wetland soil or in suspended solids deposited during tidal flooding is some orders of magnitude larger than the vegetation pool, phytoextraction is not applicable. The growth of plant species with low accumulation in aboveground tissues, e.g. Scirpus maritimus or Typha latifolia, may be preferred since this might result in lower toxic metal distribution to the wider environment.


Association between serum vitamin D levels and visceral adipose tissue among adolescents: a cross-sectional observational study in NHANES 2011-2015.
In recent years, obesity and vitamin D deficiency are more prevalent among adolescents. Improving our knowledge of the link between vitamin D and visceral adipose tissue (VAT) is essential for the health of adolescents. This study aimed to examine the connection between serum vitamin D levels and VAT mass among adolescents participating in the United States. This is a cross-sectional study that used data from the 2011 to 2015 National Health and Nutrition Examination Survey (NHANES). The connection between serum vitamin D levels and VAT was investigated using weighted multiple linear regression models. Potential nonlinear relationships were explored using smooth curve fitting. The analysis included 3171 adolescents aged 12-19 years. Vitamin D levels were shown to be inversely linked with VAT in the full-adjusted model (β = - 0.34, 95% CI: - 0.49 to - 0.19). When stratified analyses by gender, this negative relationship persisted in the girls' group (β = - 0.39, 95% CI: - 0.60 to - 0.19), but not in the boys' group (β = - 0.06, 95% CI: - 0.25 to 0.13). When stratified analysis by race, this negative relationship persisted in the Mexican American group (β = - 0.61, 95% CI: - 1.03 to - 0.19), and the non-Hispanic White group (β = - 0.27, 95% CI: - 0.54 to - 0.01), but not in the other groups. Our findings confirmed that serum vitamin D levels negatively correlated with VAT among adolescents in the United State, especially in girls, the Mexican American and non-Hispanic White. Further research is needed to determine whether increasing serum vitamin D levels decrease VAT among adolescents.


Measurement of local cerebral blood flow with [14C]iodoantipyrine in the mouse.
Local cerebral blood flow was measured in the mouse by means of the [14C]iodoantipyrine method. This method has been previously used in the monkey, dog, cat, and rat, but its application to small mammals such as the mouse requires special attention to potential sources of error. The small size of the mouse brain requires special attention to the rapid removal and freezing of the brain to minimize effects of postmortem diffusion of tracer in the tissue. Because of the relatively low diameter/length ratios of the catheters needed for arterial sampling in small animals, substantial errors can occur in the determination of the time course of the [14C]iodoantipyrine concentration in the arterial blood unless corrections for lag time and dead space washout in the catheter are properly applied. Local cerebral blood flow was measured in seven awake mice with appropriate care to minimize these sources of error. The values were found to vary from 48 ml/100 g/min in the corpus callosum to 198 ml/100 g/min in the inferior colliculus. The results demonstrate that the [14C]iodoantipyrine method can be used to measure local cerebral blood flow in the mouse and that the values in that species are, in general, somewhat higher than those in the rat.


Factors affecting sick leave prescribing by dentists.
The aim was to examine the prescribing of sick leave by dentists and factors affecting prescribing practices. A questionnaire study covering sick leave-prescribing practice during the preceding year and in response to 16 hypothetical patient cases was conducted among 1132 Finnish dentists. The effects of both dentist-related and local structural background variables on sick leave prescribing were studied. During the preceding year, dentists who were young, male, or practicing in small municipalities prescribed more days of sick leave than comparator groups. In response to the 16 hypothetical cases, dentists prescribed a mean of 31.6 days of sick leave (range, 0-98 days). The dentists with a specialty degree in oral and maxillofacial surgery prescribed longer sick leave than other dentists. Dentists working only in public dental services (PDS) and older dentists prescribed significantly less sick leave than others. There is considerable variation among dentists in sick leave-prescribing practices. The most significant factors affecting this variation are having a specialty in oral and maxillofacial surgery and working only in PDS. Further education and guidelines on dental sick leave prescribing are needed.


MTHFD2 reprograms macrophage polarization by inhibiting PTEN.
The one-carbon metabolism enzyme methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) is involved in the regulation of tumor oncogenesis and immune cell functions, but whether it can contribute to macrophage polarization remains elusive. Here, we show that MTHFD2 suppresses polarization of interferon-γ-activated macrophages (M(IFN-γ)) but enhances that of interleukin-4-activated macrophages (M(IL-4)) both in vitro and in vivo. Mechanistically, MTHFD2 interacts with phosphatase and tensin homolog (PTEN) to suppress PTEN's phosphatidylinositol 3,4,5-trisphosphate (PIP3) phosphatase activity and enhance downstream Akt activation, independent of the N-terminal mitochondria-targeting signal of MTHFD2. MTHFD2-PTEN interaction is promoted by IL-4 but not IFN-γ. Furthermore, amino acid residues (aa 215-225) of MTHFD2 directly target PTEN catalytic center (aa 118-141). Residue D168 of MTHFD2 is also critical for regulating PTEN's PIP3 phosphatase activity by affecting MTHFD2-PTEN interaction. Our study suggests a non-metabolic function of MTHFD2 by which MTHFD2 inhibits PTEN activity, orchestrates macrophage polarization, and alters macrophage-mediated immune responses.


Marijuana use and major depressive disorder are additively associated with reduced verbal learning and altered cortical thickness.
Marijuana (MJ) use and major depressive disorder (MDD) have both been associated with deficits in verbal learning and memory as well as structural brain abnormalities. It is not known if MJ use by those with MDD confers additional impairment. The goal of this study was to examine unique and combined effects of MDD and MJ use on verbal memory and brain structure. Young adults (n=141) aged 18-25 years with MJ use and no lifetime MDD (MJ, n=46), MDD and no MJ use (MDD, n=23), MJ use and lifetime MDD (MDD+MJ, n=24), and healthy controls without MDD or MJ use (CON, n=48) were enrolled. Participants completed the California Verbal Learning Test, Second Edition (CVLT-II), a measure of verbal learning and memory. A sub-sample of 82 participants also underwent a structural magnetic resonance imaging (MRI) scan. Group differences in CVLT-II performance, cortical thickness, and hippocampal volume were assessed. We found an additive effect of MDD and MJ on memory recall. Only MDD, but not MJ, was associated with poorer initial learning, fewer words recalled, more intrusion errors, and lower percent retention. There was also an additive effect of MDD and MJ use on reduced cortical thickness in the middle temporal gyrus. Findings indicate that MJ use and MDD have additive adverse associations with verbal recall and cortical thickness in the middle temporal gyrus, suggesting that MJ use among those with MDD may be contraindicated. Prospective studies are warranted to determine whether this association may be causal.


Temporal evolution of organochlorine and organophosphate pesticide residues in wells in the Akkar Region (Lebanon).
The Akkar plain is the second largest agricultural area in Lebanon. This region produces huge amount of regular crops such as maize, fruits, and vegetables. In order to protect the crops, farmers use large quantities of many pesticides (including authorized and prohibited molecules) without respecting the recommended doses. In this work, we wanted to study the evolution of OCP and OPP residues at 3-year intervals in water wells in the Akkar region. Twenty OCPs and 8 OPPs were monitored in eight wells in different villages in the plain and mountains of Akkar. Solid phase extraction (SPE) method was used for pesticide extraction, followed by gas chromatography-mass spectrometry (GC-MS) analysis. The results revealed an increasing concentration of OCPs and OPPs in groundwater over the last 3 years (between 2017 and 2019-2020). This increase in contamination is due to the uncontrolled and still unregulated (by the authorities) use of pesticides, and also to the introduction of new crops. The concentrations found in groundwater confirm that some banned pesticides are still widely used. The calculation of the theoretical pesticide intake suggests that pesticide concentrations in Akkar represent a greater health risk for the population consuming well water during the rainy season.


Computer use problems and accommodation strategies at work and home for people with systemic sclerosis: a needs assessment.
We identified the extent of the need for interventions and assistive technology to prevent computer use problems in people with systemic sclerosis (SSc) and the accommodation strategies they use to alleviate such problems. Respondents were recruited through the Scleroderma Foundation. Twenty-seven people with SSc who used a computer and reported difficulty in working completed the Computer Problems Survey. All but 1 of the respondents reported one problem with at least one equipment type. The highest number of respondents reported problems with keyboards (88%) and chairs (85%). More than half reported discomfort in the past month associated with the chair, keyboard, and mouse. Respondents used a variety of accommodation strategies. Many respondents experienced problems and discomfort related to computer use. The characteristic symptoms of SSc may contribute to these problems. Occupational therapy interventions for computer use problems in clients with SSc need to be tested.


Parametric study of a brewery effluent treatment by microalgae Scenedesmus obliquus.
This work analyses the potential of using microalgae Scenedesmus obliquus (So) for a brewery wastewater treatment and biomass production. The chemical oxygen demand (COD), total nitrogen (TN) and total carbon (TC) was followed in time, and the influence of light exposure, light intensity and culture aeration was studied. Results show that the most adequate conditions for cultivating So in this effluent are the aerated cultures, exposed to a 12h period of daily light, at 12000 Lux intensity. At these conditions it is obtained a maximum of 0.9 g of dry biomass per liter of culture, after 9 days, for a maximum reduction of 57.5% and 20.8% of COD and TN, respectively, after 14 days, and 56.9% of TC, after 13 days, corresponding to the final values of 1692 mg O(2)/L COD, 47 mg N/L TN, and 1mg C/L TC.


Correlates of protection against influenza infection in humans--on the path to a universal vaccine?
Influenza is an acute respiratory viral infection with high mutation rate and pandemic potential. Vaccination is an effective means of prevention and control of influenza, but the challenges of vaccine mismatches for the next influenza seasons and adequate global supply of influenza vaccines limit its effectiveness. Protective immunity in vaccination or natural infection is primarily mediated by antibody responses against surface proteins of influenza including haemagglutinin (HA) as the major neutralizing target, whereas strong T cell responses to internal viral proteins are associated with reduced disease severity. Recently, identification of broadly neutralizing antibodies against the conserved stem region of HA from influenza infected individuals has invigorated interest in development of a universal vaccine against different subtypes of influenza. Moreover, because of the cross-reactive nature of T cell recognition and more conserved internal antigens of influenza, strategies that boost memory T cell responses to these internal antigens may provide not only help for antibody-mediated protection but also limit the cell damage caused by viral infection directly. This is particularly important in acute infection with new pandemic viruses or antibody-escape variants where there are no pre-existing neutralizing antibodies. Here, we review the protective immune correlates against human influenza infection and discuss current status of universal influenza vaccine development.


Development of a biofeedback tilt-table for investigating orthostatic syncope in patients with spinal cord injury.
The purpose of this study was to develop a biofeedback tilt-table for automatic tilt-table training, helping patients with spinal cord injury (SCI) to recover more rapidly from orthostatic hypotension, and increasing safety to avoid syncope during training. This biofeedback tilt-table implemented automatic training maneuvers and included three closed feedback loops to monitor the acquisition of physiological signals from patients and the feedback of presyncope symptoms (PS) to regulate the angle of tilt. The results of clinical testing revealed that the mean blood pressure and oxygen saturation represented the most useful physiological signals for determining PS feedback and the quantitative criteria adopted were practicable and useful in describing the level of PS. This novel biofeedback tilt-table system offered higher patient throughput, faster training and safety in training of SCI patients to overcome their orthostatic hypotension than traditional tilt-table training, and could provide quantitative information of PS to assist medical staff in studying the mechanism of orthostatic syncope.


The TRPV4-TAZ mechanotransduction signaling axis in matrix stiffness- and TGFβ1-induced epithelial-mesenchymal transition.
The implantation of biomaterials into soft tissue leads to the development of foreign body response, a non-specific inflammatory condition that is characterized by the presence of fibrotic tissue. Epithelial-mesenchymal transition (EMT) is a key event in development, fibrosis, and oncogenesis. Emerging data support a role for both a mechanical signal and a biochemical signal in EMT. We hypothesized that transient receptor potential vanilloid 4 (TRPV4), a mechanosensitive channel, is a mediator of EMT. Normal human primary epidermal keratinocytes (NHEKs) were seeded on collagen-coated plastic plates or varied stiffness polyacrylamide gels in the presence or absence of TGFβ1, Immunofluorescence, immunoblot, and polymerase chain reaction analysis were performed to determine expression level of EMT markers and signaling proteins. Knock-down of TRPV4 function was achieved by siRNA transfection or by GSK2193874 treatment. We found that knock-down of TRPV4 blocked both matrix stiffness- and TGFβ1-induced EMT in NHEKs. In a murine skin fibrosis model, TRPV4 deletion resulted in decreased expression of the mesenchymal marker, α-SMA, and increased expression of epithelial marker, E-cadherin. Mechanistically, our data showed that: i) TRPV4 was essential for the nuclear translocation of TAZ in response to matrix stiffness and TGFβ1; ii) Antagonism of TRPV4 inhibited both matrix stiffness-induced and TGFβ1-induced expression of TAZ proteins; and iii) TRPV4 antagonism suppressed both matrix stiffness-induced and TGFβ1-induced activation of Smad2/3, but not of AKT. These data identify a novel role for TRPV4-TAZ mechanotransduction signaling axis in regulating EMT in NHEKs in response to both matrix stiffness and TGFβ1.


Unusual course of right atrial myxoma, masked by acute abdominal pain, and complicated by pulmonary embolus.
In a 17-year-old youth, exploratory laparotomy for acute abdominal pain was complicated by circulatory arrest related to pulmonary embolism. Echocardiography after resuscitation revealed a hitherto "silent" right atrial myxoma, fragmentation of which had blocked the right atrioventricular ostium, causing the pain. Operation was successful. As only a few similar cases have been reported, pulmonary embolism due to fragmentation of a right atrial myxoma may account for some unexplained sudden deaths beyond medical help.


Surgery for radiation-induced symptomatic carotid atherosclerosis.
Carotid atherosclerosis occurring secondary to cervical irradiation is known to produce stroke. Transient neurologic symptoms have necessitated surgical intervention to prevent stroke despite concern over technical problems, wound healing, operative risks, and uncertain therapeutic outcome. With this report, 26 surgical procedures in 20 patients are now documented in the literature (12 men--60%; eight women--40%). Mean age of these patients (56 years) was 10 years younger than carotid surgery patients with no prior radiation history. No relationship was noted between elevated serum cholesterol and the subsequent development of radiation-induced carotid atherosclerosis. Surgical procedures performed included carotid endarterectomy in 17 cases (65%) and arterial bypass in nine (35%). The combination of radiation therapy and previous neck surgery, including prior radical neck dissection, did not adversely influence operability. Surgical outcome was uniformly good with only one stroke (4%) documented in the perioperative period. Longer follow-up on our six cases (mean two years) disclosed neither new clinical symptoms nor the development of hemodynamically significant restenosis.


Tobacco outlet density and demographics at the tract level of analysis in Iowa: implications for environmentally based prevention initiatives.
This study assessed the geographic association between tobacco outlet density and three demographic correlates-income, race, and ethnicity-at the tract level of analysis for one county in the Midwestern United States. Data for residential census tracts in a Midwestern U.S. county were derived from year 2003 licenses for 474 tobacco outlets. Demographic variables were based on 2000 census data. Census tracts with lower median household income, higher percent of African American residents, and higher percent of Latinos residents had greater density of tobacco selling retail outlets. Areas characterized by lower income and disproportionately more African Americans and Latinos have greater physical access to tobacco products. Physical access to tobacco is a critical public-health issue because, given that smokers have been shown to be price sensitive, lowering access costs (e.g., reduced travel time) is likely to increase consumption. Findings also suggest the need for structural or environmental interventions, i.e., tobacco outlet zoning laws, to mitigate the health consequences associated with tobacco use in certain populations and geographic regions.


The effect of sediment mimicking drill cuttings on deep water rhodoliths in a flow-through system: Experimental work and modeling.
The impact of sediment coverage on two rhodolith-forming calcareous algae species collected at 100m water depth off the coast of Brazil was studied in an experimental flow-through system. Natural sediment mimicking drill cuttings with respect to size distribution was used. Sediment coverage and photosynthetic efficiency (maximum quantum yield of charge separation in photosystem II, ϕPSIImax) were measured as functions of light intensity, flow rate and added amount of sediment once a week for nine weeks. Statistical experimental design and multivariate data analysis provided statistically significant regression models which subsequently were used to establish exposure-response relationship for photosynthetic efficiency as function of sediment coverage. For example, at 70% sediment coverage the photosynthetic efficiency was reduced 50% after 1-2weeks of exposure, most likely due to reduced gas exchange. The exposure-response relationship can be used to establish threshold levels and impact categories for environmental monitoring.


Anesthetic management using a combination of anterior quadratus lumborum block and erector spinae plane block for robot-assisted partial nephrectomy: two case reports.
There has been increasing attention regarding quadratus lumborum block (QLB) and erector spinae plane block (ESPB) as effective truncal blocks. There have been reports of combined QLB and ESPB usage in hip surgery resulting in a symbiotic increase in effectiveness. However, there have been no reports regarding robot-assisted partial nephrectomy (RAPN), which requires multiple port holes ranging from near the xiphoid process to below the umbilicus. We hypothesized that the combined use of QLB and ESPB was an option for anesthesia and analgesia during RAPN. Anterior QLB and ESPB were applied to two patients undergoing scheduled RAPN. With intravenous patient-controlled analgesia, the post-surgery numerical rating scale scores were < 3/10 at rest and < 5/10 upon movement, throughout the perioperative time. The combination of QLB and ESPB could be an option for the postoperative analgesia in RAPN.


Growth Hormone Secretagogue Receptor Dimers: A New Pharmacological Target
The growth hormone secretagogue receptor (GHSR1a), the target of the ghrelin peptide, is widely distributed throughout the brain, and, while studies have often reported very low or absent levels of central ghrelin, it is now known that GHSR1a, even in the absence of a natural ligand, has physiological roles. Not only do these roles originate from the receptor's constitutive activity, but recent data indicate that GHSR1a dimerizes with a wide array of other receptors. These include the dopamine 1 receptor (D1R), the dopamine 2 receptor (D2R), the melanocortin-3 receptor (MC3R), the serotonin 2C receptor (5-HT2C), and possibly the cannabinoid type 1 receptor (CB1). Within these dimers, signaling of the protomers involved are modified through facilitation, inhibition, and even modification of signaling pathways resulting in physiological consequences not seen in the absence of these dimers. While in some cases the ghrelin peptide is not required for these modifications to occur, in others, the presence is necessary for these changes to take effect. These heterodimers demonstrate the broad array of roles and complexity of the ghrelin system. By better understanding how these dimers work, it is hoped that improved treatments for a variety of disorders, including Parkinson's disease, schizophrenia, addiction, obesity, diabetes, and more, can be devised. In this review, we examine the current state of knowledge surrounding GHSR heterodimers, and how we can apply this knowledge to various pharmacological treatments.


Prevention of tobacco use among adolescents in public schools in San Diego County, U.S.A.
Project S.H.O.U.T. (which stands for "Students Helping Others Understand Tobacco") is a tobacco-use prevention project funded by the United States National Cancer Institute for preventing the use of cigarettes and smokeless tobacco (snuff and chewing tobacco) in public schools in San Diego County, California. Based on principles of behavior modification, Project S.H.O.U.T. teaches students how to anticipate and overcome negative peer pressure to use tobacco and reinforces them for having done so. Preliminary results indicate that the prevalence of tobacco use among students receiving peer pressure resistance skills training was somewhat but not statistically significantly lower than the control group after one year of intervention. In addition, students in a third lottery condition, who were reinforced for simply not using tobacco, showed lower prevalence rates compared to controls. Additional support for the efficacy of the intervention program was provided by the refusal skills assessment test, which indicated that students exposed to the intervention were able to refuse tobacco offers more effectively than the control students. Our results indicate that tobacco use prevention experts should do more than provide decision-making and behavioral skills for resisting tobacco use, but should also reinforce the non-use of tobacco.


[Surgery simulation with a percutaneous and transpedical interbody bone grafting apparatus].
To develop a percutaneous and transpedical interbody bone grafting apparatus for vertebral bone defect reconstruction in thoracolumbar fracture correction via minimally invasive operation. The percutaneous and transpedical interbody bone grafting apparatus was designed with CAD software, and the reduction effect, range of bone grafting and surgical complications of the apparatus were investigated in adult cadaveric thoracolumbar body and with computerized surgical simulation. The self-designed apparatus was convenient for percutaneous and transpedical interbody bone grafting that did not give rise to complications. CT showed large bone grafting area with increased density in the vertebral body corrected with this apparatus. The designed apparatus allows easy manipulation and efficient bone grafting and repositioning. Minimally invasive interbody bone grafting in thoracolumbar fracture can be easily performed with proper application of the apparatus.


Molecular Profiling Reveals a Common Metabolic Signature of Tissue Fibrosis.
Fibrosis, or the accumulation of extracellular matrix, is a common feature of many chronic diseases. To interrogate core molecular pathways underlying fibrosis, we cross-examine human primary cells from various tissues treated with TGF-β, as well as kidney and liver fibrosis models. Transcriptome analyses reveal that genes involved in fatty acid oxidation are significantly perturbed. Furthermore, mitochondrial dysfunction and acylcarnitine accumulation are found in fibrotic tissues. Substantial downregulation of the PGC1α gene is evident in both in vitro and in vivo fibrosis models, suggesting a common node of metabolic signature for tissue fibrosis. In order to identify suppressors of fibrosis, we carry out a compound library phenotypic screen and identify AMPK and PPAR as highly enriched targets. We further show that pharmacological treatment of MK-8722 (AMPK activator) and MK-4074 (ACC inhibitor) reduce fibrosis in vivo. Altogether, our work demonstrate that metabolic defect is integral to TGF-β signaling and fibrosis.


[Stable isotopes of carbon and nitrogen in soil ecological studies].
The development of stable isotope techniques is one of the main methodological advances in ecology of the last decades of the 20th century. Many biogeochemical processes are accompanied by changes in the ratio between stable isotopes of carbon and nitrogen (12C/13C and 14N/15N), which allows different ecosystem components and different ecosystems to be distinguished by their isotopic composition. Analysis of isotopic composition makes it possible to trace matter and energy flows through biological systems and to evaluate the rate of many ecological processes. The main concepts and methods of stable isotope ecology and patterns of stable isotope fractionation during organic matter decomposition are considered with special emphasis on the fractionation of isotopes in food chains and the use of stable isotope studies of trophic relationships between soil animals in the field.


Development of a genetic system in chitinase-producing streptomyces and the application of an allosamidin-insensitive chitinase gene to homologous overexpression.
A transformation system for Streptomyces sp. AJ9463 strain (allosamidin producer) was successfully developed using protoplasts and a PEG-mediated method. To prepare protoplasts, the concentration of glycine and sucrose in YEME medium were optimized to 0.5% (w/v) and 34.0% (w/v), respectively. When the protoplasts of Streptomyces sp. AJ9463 were transformed with pUWL-KS, transformants could be obtained at a high efficiency of 7.0 x 10(4) transformants per microg DNA. To ensure that the transformation system worked properly, we then constructed a constitutive expression vector pYK1, in which the ermE* promoter drives transcription of the allosamidin-insensitive chitinase gene, chiIS. Although no transformant could be obtained by the genetic system using pYK1 isolated from Escherichia coli DH5alpha, pYK1 isolated from the methylase-deficient mutant E. coli SCS110, could be introduced into Streptomyces sp. AJ9463. This indicates that Streptomyces sp. AJ9463 has a methylation-specific restriction system, and that the chiIS and/or ermE* promoter region of pYK1 includes a restriction site of its endonuclease. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) indicated that pYK1 in Streptomyces sp. AJ9463 started to obviously express ChiIS from 14-h. Moreover, the pYK1-introduced strain gave a five-fold higher chitinase activity than the wild-type, suggesting that this system can be widely applied for the overexpression and gene functional analysis.


[Second allogeneic hematopoietic stem cell transplantation with reduced-intensity conditioning and donor changes in relapsed hematological malignancies after the first allogeneic transplant].
Objective: The purpose of this study was to assess the safety and efficacy of a second allogeneic hematopoietic stem cell transplantation (allo-HSCT) with reduced-intensity conditioning (RIC) in patients with hematological malignancies who had relapsed after the first allo-HSCT. Methods: Between April 2018 and June 2021, 44 patients with hematological malignancies (B-ALL 23, T-ALL/T-LBL 4, AML15, and MDS 2) were enrolled and retrospectively examined. Unrelated donors (n=12) or haploidentical donors (n=32) were used. Donors were replaced in all patients for the second allo-HSCT. Hematological and immunological germline predisposition genes and hematopoietic and immune function tests were used to select the best-related donor. Total body irradiation (TBI) /fludarabine (FLU) -based (n=38), busulfan (BU) /FLU-based (n=4), total marrow irradiation (TMI) /FLU-based (n=1), and BU/cladribine-based (n=1) were the RIC regimens used. For graft versus host disease (GVHD) prevention, cyclosporine, mycophenolate mofetil, short-term methotrexate, and ATG were used. Eighteen (40.9%) of 44 patients with gene variations for which targeted medications are available underwent post-transplant maintenance therapy. Results: The median age was 25 years old (range: 7-55). The median interval between the first and second HSCT was 19.5 months (range: 6-77). Before the second allo-HSCT, 33 (75%) of the patients were in complete remission (CR), whereas 11 (25%) were not. All patients had long-term engraftment. The grade Ⅱ-Ⅳ GVHD and severe acute GVHD rates were 20.5% and 9.1%, respectively. Chronic GVHD was found in 20.5% of limited patterns and 22.7% of severe patterns. CMV and EBV reactivation rates were 29.5% and 6.8%, respectively. Hemorrhage cystitis occurred in 15.9% of cases, grade Ⅰ or Ⅱ. The 1-yr disease-free survival (DFS), overall survival (OS), and cumulative recurrence incidence (RI) rates of all patients were 72.5% (95% CI, 54.5%-84.3%), 80.6% (95% CI, 63.4%-90.3%), and 25.1% (95% CI, 13.7%-43.2%), respectively, with a median follow-up of 14 (2-39) months. There were eight deaths (seven relapses and one infection). The rate of non-relapse mortality (NRM) was only 2.3%. The CR patients' 1-yr RI rate was significantly lower than the NR patients (16.8% vs 48.1%, P=0.026). The DFS rate in CR patients was greater than in NR patients, although there was no statistical difference (79.9% vs 51.9%, P=0.072). Univariate analysis revealed that CR before the second allo-HSCT was an important prognostic factor. Conclusion: With our RIC regimens, donor change, and post-transplant maintenance therapy, the second allo-HSCT in relapsed hematological malignancies after the first allo-HSCT is a safe and effective treatment with high OS and DFS and low NRM and relapse rate. The most important factor influencing the prognosis of the second allo-HSCT is the patient's illness condition before the transplant.


Duet: SNP-assisted structural variant calling and phasing using Oxford nanopore sequencing.
Whole genome sequencing using the long-read Oxford Nanopore Technologies (ONT) MinION sequencer provides a cost-effective option for structural variant (SV) detection in clinical applications. Despite the advantage of using long reads, however, accurate SV calling and phasing are still challenging. We introduce Duet, an SV detection tool optimized for SV calling and phasing using ONT data. The tool uses novel features integrated from both SV signatures and single-nucleotide polymorphism signatures, which can accurately distinguish SV haplotype from a false signal. Duet was benchmarked against state-of-the-art tools on multiple ONT sequencing datasets of sequencing coverage ranging from 8× to 40×. At low sequencing coverage of 8×, Duet performs better than all other tools in SV calling, SV genotyping and SV phasing. When the sequencing coverage is higher (20× to 40×), the F1-score for SV phasing is further improved in comparison to the performance of other tools, while its performance of SV genotyping and SV calling remains higher than other tools. Duet can perform accurate SV calling, SV genotyping and SV phasing using low-coverage ONT data, making it very useful for low-coverage genomes. It has great performance when scaled to high-coverage genomes, which is adaptable to various clinical applications. Duet is open source and is available at https://github.com/yekaizhou/duet .


Cooling the intermediate area of the ventral medullary surface affects tracheal responses to hypoxia.
The intermediate area of the ventral medullary surface (VMS) influences changes in airway tone caused by hypercapnia and intrapulmonary irritant receptor activation. These studies evaluated the effects of cooling the intermediate area of the VMS on the reflex hypoxic responses of the trachealis smooth muscle and of the phrenic nerve. Anesthetized, paralyzed cats were hyperventilated with 100% oxygen to produce phrenic neural apnea. Tracheal tone was measured indirectly by evaluating pressure changes in an innervated tracheal segment and the phrenic electroneurogram was determined from the central end of a cut cervical root. Switching the inspired gas to 12% O2 increased tracheal pressure of 11 of 12 cats but caused phrenic activity to reappear in only 6 of the animals. Ventilation with 6% O2 significantly increased tracheal constriction prior to phrenic activity. After intravenous administration of atropine methyl nitrate tracheal responses to hypoxia were abolished but phrenic neural responses were unaltered. Neither the tracheal pressure nor the phasic phrenic electroneurogram responded to hypoxia after cutting the carotid sinus nerves. When the intermediate area of the VMS was cooled to 20 degrees C prior to ventilation with the hypoxic gases, both tracheal and phrenic responses were significantly diminished. While the cats were hyperventilated with 6% O2, cooling of the intermediate area significantly diminished tracheal pressure and phrenic nerve activity and both returned to the same levels after rewarming. Cooling of the intermediate area blunted tracheal and phrenic responses to carotid body stimulation by NaCN. However, the appearance of tracheal constriction prior to the onset of phasic phrenic activity may suggest that increased trachealis tone may occur independent of cyclical respiratory activity.