Subthreshold excitatory activity and motoneuron discharge during REM periods of active sleep.
A striking paradox of the rapid eye movement periods of active sleep, which are typically characterized by the exacerbation of somatomotor atonia, is the occurrence of muscle twitches and jerks. The purpose of this study was to examine the specific motoneuron membrane potential processes responsible for these myoclonic patterns of activity. In lumbar motoneurons, examined intracellularly in the cat prepared for long-term study, these processes consisted of recurrent depolarizing membrane potential shifts and spontaneous action potentials that were either full-sized or of partial amplitude. In addition, the invasion of antidromically induced spikes into the soma was often blocked. Hyperpolarizing potentials were evident in the intervals between spontaneous spikes. Hyperpolarization was also observed immediately before depolarization and spike activity, in contrast to the gradual depolarization of the motoneuron membrane potential that always occurred during wakefulness. Thus, during rapid eye movement periods, in conjunction with muscle twitches and jerks, a strong excitatory input is superimposed on a background of inhibitory input. The unique patterns of membrane potential change that arise thus seem to result from the simultaneous coactivation of excitatory and inhibitory processes.


AAA+ Machines of Protein Destruction in Mycobacteria.
The bacterial cytosol is a complex mixture of macromolecules (proteins, DNA, and RNA), which collectively are responsible for an enormous array of cellular tasks. Proteins are central to most, if not all, of these tasks and as such their maintenance (commonly referred to as protein homeostasis or proteostasis) is vital for cell survival during normal and stressful conditions. The two key aspects of protein homeostasis are, (i) the correct folding and assembly of proteins (coupled with their delivery to the correct cellular location) and (ii) the timely removal of unwanted or damaged proteins from the cell, which are performed by molecular chaperones and proteases, respectively. A major class of proteins that contribute to both of these tasks are the AAA+ (ATPases associated with a variety of cellular activities) protein superfamily. Although much is known about the structure of these machines and how they function in the model Gram-negative bacterium Escherichia coli, we are only just beginning to discover the molecular details of these machines and how they function in mycobacteria. Here we review the different AAA+ machines, that contribute to proteostasis in mycobacteria. Primarily we will focus on the recent advances in the structure and function of AAA+ proteases, the substrates they recognize and the cellular pathways they control. Finally, we will discuss the recent developments related to these machines as novel drug targets.


Web-based sharing of electrocardiogram: a framework for information publishing.
Network-based data sharing is a current trend in medicine and healthcare. The search and retrieval architecture (SRA) we previously proposed for web-based sharing of electrocardiogram (ECG) facilitates the search and retrieval of ECG across hospitals via the Internet. The SRA has a triangle-like configuration including an ECG metadata registry, an ECG provider and an ECG querist. In this paper, we present a framework for ECG information publishing of an ECG provider. We also introduce a prototype of this framework, which was developed for an experimental scenario for assessment test based on MFER, an IEEE standard proposed from Japan. The assessment shows that the prototype of the framework can effectively publish the ECGs in a group of emulated MFER-conformant electrocardiographs, and the published ECGs can be successfully discovered and retrieved via the Internet.


Laparoscopic abdominal staging in locally advanced cervical cancer.
In order to determine patterns of peritoneal spread in locally advanced cervical cancer, 59 patients with previously untreated stages IB and IIA tumor size > 4 cm, IIB, III and IVA cervical cancer were considered for laparoscopic abdominal staging. Fifty-six patients (95%) were considered suitable and underwent laparoscopy. Peritoneal spread was found in 15 (27%) patients. The location was pelvic in nine (17%), extra-pelvic in one (2%), both pelvic and extra-pelvic in four (8%). Peritoneal washing was positive in five (9%) patients, being the unique site of peritoneal spread in one. Overall, 16 (29%) patients had evidence of abdominal disease. The median number of positive sites was one (range 1-4); uterine serosa was positive in nine (17%) patients, pre-vesical peritoneum in seven (13%), Douglas peritoneum in five (10%), paracolic gutter in three (6%), adnexa and omentum in two (4%), and sigmoid serosa in one (2%) patient. One operative complication occurred and all patients were discharged the day after the procedure. To date, with a median follow-up of 27 months (range 7-38), no metastasis has been detected at the trocar insertion sites. To summarize, laparoscopic staging in locally advanced cervical cancer is a safe, feasible and simple technique which is able to accurately detect abdominal disease.


[Study of the hydratation of the double-helical poly A-poly U complex by IR-spectroscopy and piezogravimetry methods].
IR-spectra of a double-helical poly A-poly U complex and coiled poly U were studied at various r.h. in a 900-3800 cm-1 region. By the method of piezomicrobalance hydration isotherms for these polynucleotides were obtained. It is concluded that, as in the DNA case, simultaneous hydration of nucleic bases the backbone of polynucleotides occurs at lower r.h., and that the poly A-poly U hydration level is higher than poly A and poly U ones separately. Drastic changes in spectral parameters of poly A poly U uracil and adenine in-ring and out-of-ring absorption bands observed in 44-76% r.h. region were interpreted as a transition to helical conformation of the complex. Calculation of resonance frequencies for these normal vibrations in the dipole-dipole approximation agrees with the experimental data.


Conversion of T4 gene 46 mutant deoxyribonucleic acid into nonviable bacteriophage particles.
The T4 amber mutant N130 in gene 46 produces, upon infection of a nonpermissive host, about 10 times more phage equivalent of deoxyribonucleic acid (DNA) than viable phage particles. Intracellular DNA is shorter than its mature phage counterpart and is converted into nonviable, light phage particles (287S) with about half of the normal DNA content, incapable either of adsorbing to or killing host bacteria. This DNA is highly unstable and breaks down, upon extraction, into subunits one-tenth the normal length of T4 DNA. Electron micrographs showed that the nonviable particles consist of normal-sized capsids of various degrees of fullness. Abnormal tail structures (naked cores with the sheaths missing) were also observed. Under conditions of arrested DNA synthesis, late messenger ribonucleic acid is produced, but some species are rare or missing, resulting in uneven transcription of the late genes. Our findings indicate that continuous DNA replication is necessary for normal transcription in T4.


CanScript, an 18-Base pair DNA sequence, boosts tumor cell-specific promoter activity: implications for targeted gene therapy.
Gene therapy protocols for the treatment of cancer often employ gene promoter sequences that are known to be over-expressed in specific tumor cell types relative to normal cells. These promoters, while specific, are often weakly active. It would be desirable to increase the activity of such promoters, while at the same time retain specificity, so that the therapeutic gene is more robustly expressed. Using a luciferase reporter DNA construct in both in vitro cell transfection assays and in vivo mouse tumor models, we have determined that in the absence of any other DNA sequence, a previously identified 18-base pair enhancer sequence called CanScript, lying upstream of the MSLN gene, has ~25% of the promoter activity of CAG, a very strong non-specific promoter/enhancer, in tumor cells in which MSLN is highly expressed. Furthermore, tandem repeat copies of CanScript enhance transcription in a dose-dependent manner and, when coupled with promoter sequences that are active in tumor cells, increase promoter activity. These findings suggest that the incorporation of CanScript into gene constructs may have application in enhancing activity of promoters used in cancer-targeting gene therapy strategies, thereby improving therapeutic efficacy.


A simple method for creating wide-field visual stimulus for electrophysiology: mapping and analyzing receptive fields using a hemispheric display.
Modern neurophysiological and psychophysical studies of vision are typically based on computer-generated stimuli presented on flat screens. While this approach allows precise delivery of stimuli, it suffers from a fundamental limitation in terms of the maximum achievable spatial coverage. This constraint becomes important in studies that require stimulation of large expanses of the visual field, such as those involving the mapping of receptive fields throughout the extent of a cortical area or subcortical nucleus, or those comparing neural response properties across a wide range of eccentricities. Here we describe a simple and highly cost-effective method for the projection of computer-generated stimuli on a hemispheric screen, which combines the advantages of computerized control and wide-field (100° × 75°) delivery, without the requirement of highly specialized hardware. The description of the method includes programming techniques for the generation of stimuli in spherical coordinates and for the quantitative determination of receptive field sizes and shapes. The value of this approach is demonstrated by quantitative electrophysiological data obtained in the far peripheral representations of various cortical areas, including automated mapping of receptive field extents in cortex that underwent plasticity following lesions.


Appraisal of vitreous syphilis antibody as a novel biomarker for the diagnosis of syphilitic uveitis: a prospective case-control study.
To determine the sensitivity and specificity of syphilis antibody tests in vitreous samples and to propose an algorithm using vitreous syphilis antibody as a supplementary test to confirm syphilitic uveitis (SU). A prospective case-control study was conducted at the Retina and Uveitis Clinic from May 2017 to January 2020. Initially, patients were classified based on syphilis serology into group 1 (positive testing) and group 2 (negative testing). Group 1 was further divided into 2 subgroups (group 1A and 1B) depending on their relevant clinical manifestations and clinical improvement. Group 2 served as a control group. Thirty-eight patients were enrolled in the study: 14 in group 1A, 5 in group 1B, and 19 in group 2B. No patient was assigned to group 2A. All patients in group 1A, representing definite SU, completed syphilis test (rapid plasma reagin [RPR], enzyme immunoassay [EIA], and fluorescent treponemal antibody-absorption [FTA-ABS]) for vitreous, and all vitreous samples yielded positive results. Of the 5 subjects in group 1B, 3 cases were considered to be not SU with different conditions, and 2 were indeterminate for SU. They presented with different features not typical of SU, and they had variable and fewer positive syphilis antibody responses. The most sensitive test for detecting syphilis antibodies in vitreous was EIA (90.9%), followed by RPR (80.0%) and FTA-ABS IgG (78.9%). EIA and FTA-ABS had the highest specificity, detecting 100% of the syphilis antibody. Vitreous analysis of syphilis antibody can serve as a supplementary test to confirm SU in selected cases as the proposed algorithm.


Motivations and Experiences Related to Women's First Same-sex Sexual Encounters.
Using women's self-identified sexual identity, the current study compares motivations for first same-sex sexual encounters as well as associated experiential outcomes. We also examine whether relations between sexual motivations and experiential outcomes differ as a function of women's sexual identity status. Participants were women (N=123), ages 18-to-29 (M=21.59, SD=3.33), who self-reported a history of same-sex sexual contact. Approximately 27% of women identified as exclusively heterosexual (i.e., EH), 35% as primarily heterosexual (i.e., 'mostly heterosexual' [MH]), and 38% as exclusively or primarily lesbian/ gay, or bisexual (i.e., LGB). Participants completed an online survey. MH and LGB women reported first same-sex sexual encounters that were more motivated by intimacy and exploration motives, relative to EH women. Compared to MH and LGB women, EH also engaged in fewer sexual activities with their first same-sex partner. Intimacy and exploration motives were related to positive experiential outcomes during first same-sex contact. Associations between motivations and experiential outcomes were not moderated by sexual identity. Findings contribute to understanding motivations and experiences related to women's first same-sex sexual encounters and show that not all women with a history of same-sex sexual contact subsequently identify with a minority sexual identity label.


A steady-state biofilm model for simultaneous reduction of nitrate and perchlorate, part 2: parameter optimization and results and discussion.
Part 1 of this work developed a steady-state, multispecies biofilm model for simultaneous reduction of nitrate and perchlorate in the H(2)-based membrane biofilm reactor (MBfR) and presented a novel method to solve it. In Part 2, the half-maximum-rate concentrations and inhibition coefficients of nitrate and perchlorate are optimized by fitting data from experiments with different combinations of influent nitrate and perchlorate concentrations. The model with optimized parameters is used to quantitatively and systematically explain how three important operating conditions (nitrate loading, perchlorate loading, and H(2) pressure) affect nitrate and perchlorate reduction and biomass distribution in these reducing biofilms. Perchlorate reduction and accumulation of perchlorate-reducing bacteria (PRB) in the biofilm are affected by four promotion or inhibition mechanisms: simultaneous use of nitrate and perchlorate by PRB and competition for H(2), the same resources in PRB, and space in a biofilm. For the hydrogen pressure evaluated experimentally, a low nitrate loading (<0.1 g N/m(2)-d) slightly promotes perchlorate removal, because of the beneficial effect from PRB using both acceptors. However, a nitrate loading of >0.6 g N/m(2)-d begins to inhibit perchlorate removal, as the competition effects become dominant.


Gender influence on defecographic abnormalities in patients with posterior pelvic floor disorders.
To compare defecographic abnormalities in symptomatic men and women and to analyze differences between men and age- and symptom-matched women. Sixty-six men (mean age: 55.4 years, range: 20-81 years) who complained of constipation and/or fecal incontinence and/or pelvic pain underwent defecography after intake of a barium meal. Radiographs were analyzed for the diagnosis of rectocele, enterocele, intussusception and perineal descent. They were compared with age- and symptom-matched women (n = 198) who underwent defecography during the same period. Normal defecography was observed in 22.7% of men vs 5.5% of women (P < 0.001). Defecography in men compared with women showed 4.5% vs 44.4% (P < 0.001) rectocele, and 10.6% vs 29.8% (P < 0.001) enterocele, respectively. No difference was observed for the diagnosis of intussusception (57.6% vs 44.9%). Perineal descent at rest was more frequent in women (P < 0.005). For the same complaint, diagnosis of defecographic abnormalities was different in men than in women: rectocele, enterocele and perineal descent at rest were observed less frequently in men than in women.


[Humoral autoimmune responses to neurospecific proteins in rats with experimental allergic encephalomyelitis and immunomodulation with allogeneic nervous tissue].
Possible immunocorrective effect of the embryonic nervous tissue (NT) and using the model of an experimental allergic encephalomyelitis (EAE) in treating patients with demyelinizing diseases is a prospective field of research. EAE was induced by immunization of rats with the mixture of the spinal cord and BCV at paw pillows. Immunocorrective effect of the allogenic NT was studied in the animals with EAE following the intraperitoneal injection of a newborn rat's NT, namely: enriched fractions of glial and neuronal cell populations. Both intact and EAE animals, treated with culture medium, were used as controls. The clinical signs appeared on 12-th day after EAE inducing. The severe clinical course of EAE was accompanied with statistically significant higher titers of the autoantibodies to MBP as compared with the mild clinical course. The results obtained evidence for the possible immunocorrective effect of enriched fractions of the newborn rat's nervous cells on EAE, and for immunosuppressive effect of the neuronal fraction of NT on the immunopathological processes in EAE.


Double oxygen-sensing vector system for robust hypoxia/ischemia-regulated gene induction in cardiac muscle in vitro and in vivo.
High-fidelity genetically encoded bio-sensors that respond to changes in cellular environmental milieu in disease offer great potential in a range of patho-physiological settings. Here a unique hypoxia-regulated vector-based system with double oxygen-sensing transcriptional elements was developed for rapid and robust hypoxia-regulated gene expression in the heart. Hypoxia-responsive cis elements were used in tandem with a single proline-modified oxygen-dependent degradation (ODD) domain of hypoxia-inducible factor-1alpha to form a double oxygen-sensing vector system (DOSVS). In adult cardiac myocytes in vitro, the DOSVS demonstrated a low background expression not different from baseline control in normoxia, and with 100% efficiency, robust, 1,000-fold induction upon hypoxia. In the heart in vivo, hypoxic and ischemic challenges elicited rapid 700-fold induction in living animals, exceeding that obtained by a high-fidelity constitutive cytomegalovirus (CMV) viral promoter. DOSVS also showed high temporal resolution in the heart in response to cyclical bouts of hypoxia in vivo. We propose that DOSVS will be valuable for a range of applications, including bio-sensing and therapeutic gene expression in the heart and other organ systems that are confronted by chronic or episodic hypoxic/ischemic stresses in vivo.


24-h esophageal pH testing in asthmatics: respiratory symptom correlation with esophageal acid events.
Gastroesophageal reflux (GER) may be a trigger for asthma and may be clinically silent. Twenty-four-hour esophageal pH testing accurately diagnoses GER in asthmatics. There are no reports correlating respiratory symptoms with esophageal acid events. This study examines the prevalence and severity of GER in asthmatics with and without reflux symptoms and examines respiratory symptom correlation with esophageal acid. All esophageal manometry and 24-h esophageal pH tests performed were reviewed in asthmatics who met entrance criteria from July 1, 1989, through November 1, 1994. GER was present if esophageal pH tests were abnormal. Results of esophageal tests were compared for asthmatics with reflux symptoms and GER and asthmatics without reflux symptoms and GER. Respiratory symptoms correlated with esophageal acid events if the esophageal pH was < 4 simultaneously with the respiratory event or within 5 min before its onset. Of 199 asthmatics who qualified for analysis, 164 (82%) had reflux symptoms. The results of 24-h esophageal pH tests were abnormal in 118 of 164 asthmatics with reflux symptoms (72%), compared with 10 of 35 asthmatics without reflux symptoms (29%). Among asthmatics with GER, 119 of 151 respiratory symptoms (78.8%) were associated with esophageal acid. Seventy-six of 84 reported coughs (90.5%) were associated with esophageal acid. Theophylline did not alter esophageal parameters. There is a strong correlation between esophageal acid events and respiratory symptoms in asthmatics with GER. Respiratory symptom correlation with esophageal acid events further supports that GER may be a trigger for asthma.


Cilostazol reduces the progression of carotid intima-media thickness without increasing the risk of bleeding in patients with acute coronary syndrome during a 2-year follow-up.
Cilostazol, a phosphodiesterase III inhibitor, is known to have anti-proliferative activity. We investigated the effects of cilostazol 200 mg, in addition to aspirin 100 mg and clopidogrel 75 mg, on carotid intima-media thickness (IMT) progression during a 2-year follow-up period in patients with acute coronary syndrome (ACS) requiring stent implantation. Patients with ACS (n = 130) were randomly assigned to the cilostazol group (n = 64) or the control group (n = 66). Longitudinal images of left and right carotid IMT were measured at baseline, at 6, 12, and 24 months using a 10-MHz linear vascular probe. The primary endpoint was to compare the changes in maximum carotid IMT at 2 years. Other parameters such as inflammatory markers [interleukin (IL)-6, tumor necrosis factor (TNF)-α, C-reactive protein (CRP), and adiponectin] and bleeding risk were also compared. The carotid IMT showed no significant progression from baseline in the cilostazol group compared to significant progression in the control group at 12 months (0.78 ± 0.38 and 0.85 ± 0.41 mm, p = 0.034, respectively) and 24 months (0.82 ± 0.41 and 0.96 ± 0.39 mm, p = 0.022, respectively). Major bleeding (p = 1.00), minor bleeding (p = 0.68), and total bleeding rates (p = 0.74) were similar between the two groups during the 2-year follow-up. Decreases from baseline in IL-6 (-2.79 ± 2.83 and -2.14 ± 3.36 pg/ml, p = 0.010, respectively) and TNF-α (-2.81 ± 1.97 and -2.21 ± 2.68 pg/ml, p = 0.029, respectively) were significantly greater in the cilostazol group than the control group during the follow-up. Cilostazol treatment, with greater anti-inflammatory effect, inhibited the progression of carotid IMT without increasing the risk of bleeding in patients with ACS during the 2-year follow-up.


Four minutes of capacitive and resistive electric transfer therapy increased jump performance.
Capacitive and resistive electric transfer (CRET) therapy can improve flexibility and increase muscle activity and may be useful as a warm-up technique. This study examined the effects of short-time CRET on jump performance. Thirty healthy men (age range, 20-40 years) were randomly divided into passive (n = 15) and active (n = 15) warm-up groups. The participants and statisticians were blinded to the participant allocation. The passive warm-up group underwent 4 min of CRET therapy on their posterior lower legs. The active warm-up group performed stretching and jogging for 4 min. Calf muscle temperature and rebound jump (RJ) index were measured before and after the intervention. The mean (± standard deviation) muscle temperature increased by 2.0 ± 0.5°C and 1.4 ± 0.6°C in the passive and active warm-up groups, respectively (p < 0.05). RJ index increased significantly in both groups (p < 0.05). Therefore, passive warm-up using CRET may help avoid energy loss while increasing the muscle temperature in a short time when compared with traditional active warm-up techniques.


Environmental drivers of ectomycorrhizal communities in Europe's temperate oak forests.
Ectomycorrhizal fungi are major ecological players in temperate forests, but they are rarely used in measures of forest condition because large-scale, high-resolution, standardized and replicated belowground data are scarce. We carried out an analysis of ectomycorrhizas at 22 intensively monitored long-term oak plots, across nine European countries, covering complex natural and anthropogenic environmental gradients. We found that at large scales, mycorrhizal richness and evenness declined with decreasing soil pH and root density, and with increasing atmospheric nitrogen deposition. Shifts in mycorrhizas with different functional traits were detected; mycorrhizas with structures specialized for long-distance transport related differently to most environmental variables than those without. The dominant oak-specialist Lactarius quietus, with limited soil exploration abilities, responds positively to increasing nitrogen inputs and decreasing pH. In contrast, Tricholoma, Cortinarius and Piloderma species, with medium-distance soil exploration abilities, show a consistently negative response. We also determined nitrogen critical loads for moderate (9.5-13.5 kg N/ha/year) and drastic (17 kg N/ha/year) changes in belowground mycorrhizal root communities in temperate oak forests. Overall, we generated the first baseline data for ectomycorrhizal fungi in the oak forests sampled, identified nitrogen pollution as one of their major drivers at large scales and revealed fungi that individually and/or in combination with others can be used as belowground indicators of environmental characteristics.


Thyroid Warthin-Like Cancer Concurrent With Multiple Sclerosis: A Case Report.
Warthin like-papillary thyroid cancer (WL-PTC) is a rare malignancy; it is difficult to distinguish preoperatively a WLPTC from a classic PTC. Often it is associated with Hashimoto thyroiditis (HT) that determines a better prognosis with a very low probability of recurrence. The case concerns a 43-year-old female with a single thyroid nodule suspected for cancer; and she had multiple sclerosis (MS) from the age of 19. Thyroid hormone levels were normal such as thyroid antibodies. Total thyroidectomy with lymphadenectomy of central compartment was performed. Histological examination revealed a Warthin like-PTC without Hashimoto thyroiditis. The mechanisms involved in pathogenesis of thyroid cancer in patients with autoimmune disease are not completely clear. We hypothesized that not only a local autoimmune response, such as HT, could contribute to the determination of this type of cancer but also a systemic autoimmune disease such as MS.


Cloning and molecular analysis of the Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyalkanoate) biosynthesis genes in Pseudomonas sp. strain 61-3.
Two types of polyhydroxyalkanoate (PHA) biosynthesis gene loci (phb and pha) of Pseudomonas sp. strain 61-3, which produces a blend of poly(3-hydroxybutyrate) [P(3HB)] homopolymer and a random copolymer (poly(3-hydroxybutyrate-co-3-hydroxyalkanoate) [P(3HB-co-3HA]) consisting of 3HA units of 4 to 12 carbon atoms, were cloned and analyzed at the molecular level. In the phb locus, three open reading frames encoding polyhydroxybutyrate (PHB) synthase (PhbCPs), beta-ketothiolase (PhbAPs), and NADPH-dependent acetoacetyl coenzyme A reductase (PhbBPs) were found. The genetic organization showed a putative promoter region, followed by phbBPs-phbAPs-phbCPs. Upstream from phbBPs was found the phbRPs gene, which exhibits significant similarity to members of the AraC/XylS family of transcriptional activators. The phbRPs gene was found to be transcribed in the opposite direction from the three structural genes. Cloning of phbRPs in a relatively high-copy vector in Pseudomonas sp. strain 61-3 elevated the levels of beta-galactosidase activity from a transcriptional phb promoter-lacZ fusion and also enhanced the 3HB fraction in the polyesters synthesized by this strain, suggesting that PhbRPs is a positive regulatory protein controlling the transcription of phbBACPs in this bacterium. In the pha locus, two genes encoding PHA synthases (PhaC1Ps and PhaC2Ps) were flanked by a PHA depolymerase gene (phaZPs), and two adjacent open reading frames (ORF1 and phaDPs), and the gene order was ORF1, phaC1Ps, phaZPs, phaC2Ps, and phaDPs. Heterologous expression of the cloned fragments in PHA-negative mutants of Pseudomonas putida and Ralstonia eutropha revealed that PHB synthase and two PHA synthases of Pseudomonas sp. strain 61-3 were specific for short chain length and both short and medium chain length 3HA units, respectively.


In vitro chemoseparation of leukemic cells from murine bone marrow using VP16-213: importance of stem cell assays.
The use of chemopurified autologous bone marrow (BM) is being explored as a transplant source for patients with leukemia who do not have a HLA-matched donor. Because stem cell assays have not previously been found to predict engraftment after transplantation, the optimal drug(s) and drug concentrations have not been determined. To determine the effectiveness of the podophyllotoxin derivative, VP16, and the value of stem cell assays in chemopurification studies, a murine model using the C57B1/6 mouse and its syngeneic leukemia EL-4 was developed. Kill of committed (CFU-C) and pluripotent (CFU-S) hematopoietic stem cells and tumor (tCFU) stem cells after a 1-h exposure to VP16 was first determined. A marked kill differential of tCFU compared to that of the CFU-C/S populations was found, with no tCFU surviving at VP16 concentrations greater than 30 micrograms/ml. No kill differential of CFU-C versus CFU-S was seen at VP16 doses greater than 10 micrograms/ml. All mice transplanted with a mixture of 25 X 10(6) BM cells and 1 X 10(7) EL-4 cells treated in vitro with 40 micrograms/ml of VP16 died of their tumors. However by reducing the tumor burden to 1 X 10(6) EL-4 and 5 X 10(6) EL-4, nine of ten and four of six of the mice, respectively, survived 90+ days tumor free. On the basis of survival data, it was found that engraftment of VP16-treated BM was directly proportional to the product of the degree of CFU-S inhibition and BM cell inoculum, i.e., the number of viable CFU-S transplanted. The maximum VP16 concentration that led to predictable engraftment at BM doses less than 25 X 10(6) cells was 55 micrograms/ml. Thus, 15 mice were transplanted with 1 X 10(7) BM cells and 1 X 10(7) EL-4 cells, incubated with 55 micrograms/ml of VP16; 13 out of 15 survived tumor free for 90+ days. When VP16 was used as a BM chemopurification agent, up to 50% of contaminating tumor cells were eliminated from BM suspensions without affecting engraftment after transplantation. Because stem cell inhibition predicted engraftment, drug concentrations that maximized tumor cell kill could be chosen.


Adult and fetal human mesangial cells interact with specific laminin domains.
Mesangial cells are centrally located pericytes in the renal glomerulus. They are surrounded by an extracellular matrix and directly contact the glomerular basement membrane in vivo. Because these interactions are critical for renal development and function, we have studied human mesangial cell interactions with laminin, a major adhesive component of basement membranes present in the extracellular matrix of the mesangium. Human fetal and adult mesangial cell attachment was stimulated by both laminin and the laminin-derived synthetic peptides YIGSR-NH2, CQAGTFALRGDNPQG-NH2, and CIKVAVS-NH2. Furthermore, mesangial cells spread on laminin as well as on both the RGD-containing and CIKVAVS peptides. When added in solution, all three peptides inhibited mesangial cell attachment to laminin, and the latter two peptides inhibited mesangial cell spreading on laminin. Laminin affinity column chromatography demonstrated several low-molecular-mass laminin-binding proteins ranging from between 35 and 42 kDa, which predominated in fetal mesangial cells, whereas a higher molecular mass laminin-binding protein of 65 kDa was predominant in adult mesangial cells. Western blot analysis with an anti-32-kDa laminin-binding protein antibody showed increased expression of both 31- and 42-kDa proteins in fetal mesangial cells when compared with the adult. The antisera to the 32-kDa laminin-binding protein also inhibited fetal mesangial spreading on the CIKVAVS peptide. Western blot analysis with an anti-67-kDa laminin-binding protein antibody revealed a 110-kDa protein in adult mesangial cells that was not present in fetal mesangial cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Characterization of a plant-transformation-ready large-insert BIBAC library of Arabidopsis and bombardment transformation of a large-insert BIBAC of the library into tobacco.
Plant-transformation-ready, large-insert binary bacterial artificial chromosome (BIBAC) libraries are of significance for functional and network analysis of large genomic regions, gene clusters, large-spanning genes, and complex loci in the post-genome era. Here, we report the characterization of a plant-transformation-ready BIBAC library of the sequenced Arabidopsis genome for which such a library is not available to the public, the transformation of a large-insert BIBAC of the library into tobacco by biolistic bombardment, and the expression analysis of its containing genes in transgenic plants. The BIBAC library was constructed from nuclear DNA partially digested with BamHI in the BIBAC vector pCLD04541. It contains 6144 clones and has a mean insert size of 108 kb, representing 5.2× equivalents of the Arabidopsis genome or a probability of greater than 99% of obtaining at least one positive clone from the library using a single-copy sequence as a probe. The transformation of the large-insert BIBAC and analyses of the transgenic plants showed that not only did transgenic plants have intact BIBAC DNA, but also could the BIBAC be transmitted stably into progenies and its containing genes be expressed actively. These results suggest that the large-insert BIBAC library, combined with the biolistic bombardment transformation method, could provide a useful tool for large-scale functional analysis of the Arabidopsis genome sequence and applications in plant-molecular breeding.


Characterizing the Late Pleistocene MSA Lithic Technology of Sibudu, KwaZulu-Natal, South Africa.
Studies of the African Middle Stone Age (MSA) have become central for defining the cultural adaptations that accompanied the evolution of modern humans. While much of recent research in South Africa has focused on the Still Bay and Howiesons Poort (HP), periods following these technocomplexes were often neglected. Here we examine lithic assemblages from Sibudu that post-date the HP to further the understanding of MSA cultural variability during the Late Pleistocene. Sibudu preserves an exceptionally thick, rich, and high-resolution archaeological sequence that dates to ∼ 58 ka, which has recently been proposed as type assemblage for the "Sibudan". This study presents a detailed analysis of the six uppermost lithic assemblages from these deposits (BM-BSP) that we excavated from 2011-2013. We define the key elements of the lithic technology and compare our findings to other assemblages post-dating the HP. The six lithic assemblages provide a distinct and robust cultural signal, closely resembling each other in various technological, techno-functional, techno-economic, and typological characteristics. These results refute assertions that modern humans living after the HP possessed an unstructured and unsophisticated MSA lithic technology. While we observed several parallels with other contemporaneous MSA sites, particularly in the eastern part of southern Africa, the lithic assemblages at Sibudu demonstrate a distinct and so far unique combination of techno-typological traits. Our findings support the use of the Sibudan to help structuring this part of the southern African MSA and emphasize the need for further research to identify the spatial and temporal extent of this proposed cultural unit.


Methotrexate accumulates to similar levels in animals transplanted with normal versus drug-resistant transgenic marrow.
Gene transfer and expression of methotrexate (MTX)-resistant variants of dihydrofolate reductase (DHFR) in normal hematopoietic cells is a potential strategy to permit administration of larger doses of MTX by alleviating drug toxicity in normal cells and tissues that are drug sensitive. We have previously demonstrated that transplantation of marrow from transgenic mice expressing drug-resistant DHFRs conferred upon normal recipient animals resistance to MTX at levels that are usually toxic for hematopoietic and gastrointestinal (GI) tissues. One explanation for the observed protection from GI toxicity by drug-resistant marrow is that MTX could be cleared more rapidly in animals maintaining a more healthy hematopoietic system. To evaluate this possibility, we carried out MTX pharmacokinetic studies in mice that received transplanted transgenic marrow expressing either of two different DHFR variants, administering increasing doses of MTX up to 4 mg/kg/day. Animals received i.p. injection precisely every 24 h. Every 4 days, three animals from each group were sacrificed, and their plasma and intestines were assayed for MTX. Animals transplanted with transgenic Arg-22 DHFR drug-resistant marrow maintained hematocrit levels that were about 4-fold higher at 3 weeks after transplant than those of untreated animals or animals that received normal marrow cells. Animals that received normal marrow did not survive beyond 25 days and did not accumulate higher levels of MTX than animals that received a transgenic marrow transplant. Untreated animals exhibited a higher rate of survival (36 days) but again did not accumulate higher levels of MTX than the transgenic marrow recipients. When the experiment was repeated using transgenic Tyr-22 DHFR marrow, the levels of MTX in the plasma or GI tissues did not differ significantly between groups. Intestinal concentrations of MTX in both experiments were about 4-5-fold higher than those in the plasma. These results indicate that protection from MTX toxicity conferred by expression of drug-resistant DHFR activity in the marrow is not the result of a higher rate of MTX clearance from the circulation in comparison with control animals but a true resistance of hematopoietic and GI tissues to MTX. The maintenance of antifolate levels in animals protected from MTX toxicity implies that this procedure should not compromise the antitumor efficacy of MTX.


A base-excision DNA-repair protein finds intrahelical lesion bases by fast sliding in contact with DNA.
A central mystery in the function of site-specific DNA-binding proteins is the detailed mechanism for rapid location and binding of target sites in DNA. Human oxoguanine DNA glycosylase 1 (hOgg1), for example, must search out rare 8-oxoguanine lesions to prevent transversion mutations arising from oxidative stress. Here we report high-speed imaging of single hOgg1 enzyme molecules diffusing along DNA stretched by shear flow. Salt-concentration-dependent measurements reveal that such diffusion occurs as hOgg1 slides in persistent contact with DNA. At near-physiologic pH and salt concentration, hOgg1 has a subsecond DNA-binding time and slides with a diffusion constant as high as 5 x 10(6) bp(2)/s. Such a value approaches the theoretical upper limit for one-dimensional diffusion and indicates an activation barrier for sliding of only 0.5 kcal/mol (1 kcal = 4.2 kJ). This nearly barrierless Brownian sliding indicates that DNA glycosylases locate lesion bases by a massively redundant search in which the enzyme selectively binds 8-oxoguanine under kinetic control.


Peptide identification by database search of mixture tandem mass spectra.
In high-throughput proteomics the development of computational methods and novel experimental strategies often rely on each other. In certain areas, mass spectrometry methods for data acquisition are ahead of computational methods to interpret the resulting tandem mass spectra. Particularly, although there are numerous situations in which a mixture tandem mass spectrum can contain fragment ions from two or more peptides, nearly all database search tools still make the assumption that each tandem mass spectrum comes from one peptide. Common examples include mixture spectra from co-eluting peptides in complex samples, spectra generated from data-independent acquisition methods, and spectra from peptides with complex post-translational modifications. We propose a new database search tool (MixDB) that is able to identify mixture tandem mass spectra from more than one peptide. We show that peptides can be reliably identified with up to 95% accuracy from mixture spectra while considering only a 0.01% of all possible peptide pairs (four orders of magnitude speedup). Comparison with current database search methods indicates that our approach has better or comparable sensitivity and precision at identifying single-peptide spectra while simultaneously being able to identify 38% more peptides from mixture spectra at significantly higher precision.


Incidence of Shoulder Labral Pathology in United States Military Members.
Shoulder injuries account for approximately 8% to 24% of all musculoskeletal injuries in the military. Recently, a change was made to service-specific physical fitness tests. Knowledge of relative shoulder labral injury rates before and after this change would help guide future directions and preventive strategies. However, we found no previous literature evaluating the rates of labral injury among United States Military branch personnel by enlistment status (enlisted versus officer), gender, age, or race. In this retrospective epidemiological study, we queried the Defense Medical Epidemiology Database for International Classification of Disease, Tenth Revision (ICD-10) code S43.43 (superior glenoid labrum lesion) to determine the total number of patients with a shoulder labral injury from 2016 to 2019. Results were assessed for demographic associations of shoulder labral injury with branch, service occupation, rank, gender, race, and age. Queries were limited to first-time occurrences and ambulatory data only. Overall, our study found the incidence of shoulder labral injuries to be largely conserved each year from 2016 to 2019 (3.22-3.35/1000/year). Incidence of labral injury was highest in males, White service members, the junior enlisted, the Army service branch, ages 20 to 29, and enlisted non-combat personnel. With knowledge of injury patterns in specific military populations, initiatives may be taken to identify at-risk service members with the goal of informing future preventive strategies.


Barriers to Obtaining a Timely Diagnosis in Human Papillomavirus-Associated Oropharynx Cancer.
Failure to recognize symptoms of human papillomavirus-associated oropharyngeal squamous cell carcinoma (HPV(+)OPSCC) at presentation can delay diagnosis and treatment. This study aims to identify patient factors and provider patterns that contribute to delayed diagnosis. Retrospective case series. Tertiary care center. Patients with HPV(+)OPSCC receiving intent-to-cure treatment from 2006 to 2016. Clinical data, workup, and care timelines were abstracted. Univariate and multivariable linear regressions were performed to determine associations. Of 703 included patients, 627 (89%) were male, and mean (SD) age at diagnosis was 59 (9) years. The mean (SD) delay to diagnosis was 148.8 (243.51) days, with an average delay of 63 (154.91) days from symptom onset to first presentation and 82.8 (194.25) days from first presentation to diagnosis. Most patients visited at least 2 providers (n = 546, 78%) before diagnosis and saw their primary care physician at first presentation (n = 496, 71%). The most common imaging and biopsy obtained before diagnosis was neck computed tomography (n = 391, 56%) and neck fine-needle aspiration (n = 423, 60%), respectively. On multivariable linear regression, being a homemaker, being a current smoker, seeing 3 or more providers, and getting a magnetic resonance imaging scan were associated with significant delays in diagnosis (P < .01, all). Treatment with antibiotics and a suspicion for HPV(+)OPSCC at first presentation were associated with decreased delays in diagnosis (P < .01, both). Patient delays in seeking medical attention and provider delays in recognizing the appropriate diagnosis both contribute to delays of care in HPV(+)OPSCC. Improved patient and provider education is necessary to expedite the diagnosis of HPV(+)OPSCC.


Intrathyroidal ectopic thymic tissue mimicking thyroid cancer in children.
To increase awareness of the important differential diagnosis between thyroid cancer and intrathyroidal ectopic thymic tissue when performing neck ultrasound (US) in pediatric patients. We report 2 cases of intrathyrodal thymic tissue that were initially referred as possible papillary thyroid cancer (PTC). US and cytologic features of these lesions are described, and the distinguishing US characteristics of intrathyroidal thymic tissue versus PTC are discussed. Two pediatric patients who underwent thyroid US were diagnosed with thyroid nodules suggestive of PTC. The suspected lesions in both patients exhibited an echo texture identical to the normal thymus gland. Fine-needle aspirate (FNA) biopsy of the nodule in the first case revealed numerous lymphocytes with positive immunocytochemical staining for CD3, confirming a T cell phenotype. The second patient underwent follow-up US imaging 8 months later that showed stability of the intrathyroidal nodule. Intrathyroidal ectopic thymic tissue can be mistaken for PTC as both conditions have similar US features. Increased awareness of this unique mimicry may help to avoid unnecessary invasive investigations and surgery in these young patients.


Lithium protection of phencyclidine-induced neurotoxicity in developing brain: the role of phosphatidylinositol-3 kinase/Akt and mitogen-activated protein kinase kinase/extracellular signal-regulated kinase signaling pathways.
Phencyclidine (PCP) and other N-methyl-D-aspartate (NMDA) receptor antagonists have been shown to be neurotoxic to developing brains and to result in schizophrenia-like behaviors later in development. Prevention of both effects by antischizophrenic drugs suggests the validity of PCP neurodevelopmental toxicity as a heuristic model of schizophrenia. Lithium is used for the treatment of bipolar and schizoaffective disorders and has recently been shown to have neuroprotective properties. The present study used organotypic corticostriatal slices taken from postnatal day 2 rat pups to investigate the protective effect of lithium and the role of the phosphatidylinositol-3 kinase (PI-3K)/Akt and mitogen-activated protein kinase kinase/extracellular signal-regulated kinase (MEK/ERK) pathways in PCP-induced cell death. Lithium pretreatment dose-dependently reduced PCP-induced caspase-3 activation and DNA fragmentation in layers II to IV of the cortex. PCP elicited time-dependent inhibition of the MEK/ERK and PI-3K/Akt pathways, as indicated by dephosphorylation of ERK1/2 and Akt. The proapoptotic factor glycogen synthase kinase (GSK)-3beta was also dephosphorylated at serine 9 and thus activated. Lithium prevented PCP-induced inhibition of the two pathways and activation of GSK-3beta. Furthermore, blocking either PI-3K/Akt or MEK/ERK pathway abolished the protective effect of lithium, whereas inhibiting GSK-3beta activity mimicked the protective effect of lithium. However, no cross-talk between the two pathways was found. Finally, specific GSK-3beta inhibition did not prevent PCP-induced dephosphorylation of Akt and ERK. These data strongly suggest that the protective effect of lithium against PCP-induced neuroapoptosis is mediated through independent stimulation of the PI-3K/Akt and ERK pathways and suppression of GSK-3beta activity.


Targeting AhR as a Novel Therapeutic Modality against Inflammatory Diseases.
For decades, activation of Aryl Hydrocarbon Receptor (AhR) was excluded from consideration as a therapeutic approach due to the potential toxic effects of AhR ligands and the induction of the cytochrome P450 enzyme, Cyp1a1, following AhR activation. However, it is now understood that AhR activation not only serves as an environmental sensor that regulates the effects of environmental toxins, but also as a key immunomodulator where ligands induce a variety of cellular and epigenetic mechanisms to attenuate inflammation. Thus, the emergence of further in-depth research into diverse groups of compounds capable of activating this receptor has prompted reconsideration of its use therapeutically. The aim of this review is to summarize the body of research surrounding AhR and its role in regulating inflammation. Specifically, evidence supporting the potential of targeting this receptor to modulate the immune response in inflammatory and autoimmune diseases will be highlighted. Additionally, the opportunities and challenges of developing AhR-based therapies to suppress inflammation will be discussed.


The impact of educational and environmental interventions in Dutch worksite cafeterias.
Environmental interventions such as labeling and an increased availability of healthy foods may help consumers to meet guidelines for a healthy diet. This article describes a study into the effectiveness of two environmental programs to be used in worksite cafeterias along with an educational program. The aim of the interventions was to reduce fat intake, and to increase fruit and vegetable intake. In the labeling program, low-fat products were labeled. The food supply program comprised an increased availability of low-fat products and fruits and vegetables in worksite cafeterias. The educational program consisted of information about healthy nutrition through brochures, table tents, a self-help manual and posters. The design consisted of a pre-test-post-test experimental control group design, with four conditions: the educational program; the food supply program plus educational program; the labeling program plus educational program; and a control group. Seventeen worksites were randomly assigned to one of the four research conditions. Total fat, fruit and vegetable intake was measured with a quantitative, self-administered food frequency questionnaire (35 questions). Intake during lunch was measured by asking respondents to write down which food items they had purchased during their last lunch in the cafeteria. Furthermore, sales data for some targeted product categories were collected (milk, butter, cheese, meat products, desserts). For the whole study population, no significant effects on consumption data were found for any of the programs. The data showed a beneficial and significant treatment effect of the labeling program on total fat intake for respondents who believed they ate a high-fat diet. Sales data revealed a significant effect of the labeling program on desserts, but not for the other products.


Identifying solubility-promoting buffers for intrinsically disordered proteins prior to purification.
Intrinsically disordered proteins are anticipated to be more prone to aggregation than folded, stable proteins. Chemical additives included in the buffer can help maintain proteins in a soluble, monomeric state. However, the array of chemicals that impact protein solubility is staggering, precluding iterative testing of chemical conditions during purification. Herein, we describe a filter-based aggregation assay to rapidly identify chemical additives that maintain solubility for a protein of interest. A hierarchical approach to buffer selection is provided, in which the type of chemical which best improves solubility is first determined, followed by identifying the optimal chemical and its most effective concentration. Finally, combinations of chemical additives can be assessed if necessary. Although this assay can be applied to purified protein, partially purified protein, or aggregated protein, this protocol specifically details the use of this assay for crude cell lysate. This approach allows identification of solubility-promoting buffers prior to the initial protein purification.


General practitioners and work in the Third World.
In recent years the number of general practitioners who have worked in the third world before entering general practice has fallen. The reasons for this are not clear but may include worries about future career prospects. Ninety four doctors who had entered general practice since 1984, after previously working in the third world, completed a questionnaire about their career experience and views about the value of such work. They were generally widely experienced and well-qualified and work abroad had not apparently harmed their careers, rather, many believed it had enhanced it. Work in the usually arduous conditions of poor countries was often considered by the respondents to lead to a wider perspective, increased maturity, confidence, self-reliance, adaptability and initiative. Doctors who are interested and suitable for work in the third world prior to entering general practice should be encouraged to pursue this possibility.


Gazing down increases standing and walking postural steadiness.
When walking on an uneven surface or complex terrain, humans tend to gaze downward. This behaviour is usually interpreted as an attempt to acquire useful information to guide locomotion. Visual information, however, is not used exclusively for guiding locomotion; it is also useful for postural control. Both locomotive and postural control have been shown to be sensitive to the visual flow arising from the respective motion of the individual and the three-dimensional environment. This flow changes when a person gazes downward and may present information that is more appropriate for postural control. To investigate whether downward gazing can be used for postural control, rather than exclusively for guiding locomotion, we quantified the dynamics of standing and walking posture in healthy adults, under several visual conditions. Through these experiments we were able to demonstrate that gazing downward, just a few steps ahead, resulted in a steadier standing and walking posture. These experiments indicate that gazing downward may serve more than one purpose and provide sufficient evidence of the possible interplay between the visual information used for guiding locomotion and that used for postural control. These findings contribute to our understanding of the control mechanism/s underlying gait and posture and have possible clinical implications.


First report of a leopard (Panthera pardus)-bonobo (Pan paniscus) encounter at the LuiKotale study site, Democratic Republic of the Congo.
Predation is a major cause of mortality in non-human primates, and considered a selective force in the evolution of primate societies. Although larger body size is considered as protection against predation, evidence for predation on great apes by carnivores comes from chimpanzees (Pan troglodytes), gorillas (Gorilla gorilla), and orangutans (Pongo spp.). Here, we describe the first encounter between wild bonobos (Pan paniscus) and a leopard (Panthera pardus). A single leopard was confronted by a group of habituated bonobos for three hours. Two adult males and one adolescent female bonobo actively harassed the leopard, which remained still for most of the encounter and reacted only to close approaches by bonobos. While no predation was observed, their behaviours confirm that bonobos perceive leopards as potential predators. Our report adds novel information to descriptions from other African ape species, and sheds light on the behavioural repertoire of bonobos' anti-predation strategies. For future investigations, we suggest tagging leopards to remotely monitor their movements and allow assessment of encounter rates as one of several factors influencing predation pressure.


[Source of light-evoked cortical activity after destruction of the main visual pathway].
Electrical responses to flashes and single electrical stimuli applied to optical nerve were recorded from the visual and sensomotor cortex, the lateral geniculate nucleus, the hypothalamus and the caudate nucleus in normal cats and after bilateral lesion of the specific visual pathways. The light-imduced EPs were preserved after transection of the optical tracts and after lesion of the lateral geniculate nuclei. The EPs to flashes were absent after transection of the optical nerves. After lesion of the lateral geniculate nuclei, sparing all optical tract connections to the colliculi superior, activation of the EEG was possible with strong light flashes. The significance of the retino-hypothalamic and retino-collicular connection for formation of the light-induced cortical responses, is discussed.


Vitamin B12 may inhibit RNA-dependent-RNA polymerase activity of nsp12 from the SARS-CoV-2 virus.
SARS-CoV-2 is the causative agent for the ongoing COVID19 pandemic, and this virus belongs to the Coronaviridae family. Like other members of this family, the virus possesses a positive-sense single-stranded RNA genome. The genome encodes for the nsp12 protein, which houses the RNA-dependent-RNA polymerase (RdRP) activity responsible for the replication of the viral genome. A homology model of nsp12 was prepared using the structure of the SARS nsp12 (6NUR) as a model. The model was used to carry out in silico screening to identify molecules among natural products, or Food and Drug Administration-approved drugs that can potentially inhibit the activity of nsp12. This exercise showed that vitamin B12 (methylcobalamin) may bind to the active site of the nsp12 protein. A model of the nsp12 in complex with substrate RNA and incoming NTP showed that vitamin B12 binding site overlaps with that of the incoming nucleotide. A comparison of the calculated energies of binding for RNA plus NTP and methylcobalamin suggested that the vitamin may bind to the active site of nsp12 with significant affinity. It is, therefore, possible that methylcobalamin binding may prevent association with RNA and NTP and thus inhibit the RdRP activity of nsp12. Overall, our computational studies suggest that methylcobalamin form of vitamin B12 may serve as an effective inhibitor of the nsp12 protein.


Links between Adolescents' Closeness to Adoptive Parents and Attachment Style in Young Adulthood.
This study examined whether adolescents' closeness to adoptive parents (APs) predicted attachment styles in close relationships outside their family during young adulthood. In a longitudinal study of domestic infant adoptions, closeness to adoptive mother and adoptive father was assessed in 156 adolescents (M = 15.7 years). Approximately nine years later (M = 25.0 years), closeness to parents was assessed again as well as attachment style in their close relationships. Multilevel modeling was used to predict attachment style in young adulthood from the average and discrepancy of closeness to adolescents' adoptive mothers and fathers and the change over time in closeness to APs. Less avoidant attachment style was predicted by stronger closeness to both APs during adolescence. Increased closeness to APs over time was related to less anxiety in close relationships. Higher closeness over time to either AP was related to less avoidance and anxiety in close relationships.


Mangroves are an overlooked hotspot of insect diversity despite low plant diversity.
The world's fast disappearing mangrove forests have low plant diversity and are often assumed to also have a species-poor insect fauna. We here compare the tropical arthropod fauna across a freshwater swamp and six different forest types (rain-, swamp, dry-coastal, urban, freshwater swamp, mangroves) based on 140,000 barcoded specimens belonging to ca. 8500 species. We find that the globally imperiled habitat "mangroves" is an overlooked hotspot for insect diversity. Our study reveals a species-rich mangrove insect fauna (>3000 species in Singapore alone) that is distinct (>50% of species are mangrove-specific) and has high species turnover across Southeast and East Asia. For most habitats, plant diversity is a good predictor of insect diversity, but mangroves are an exception and compensate for a comparatively low number of phytophagous and fungivorous insect species by supporting an unusually rich community of predators whose larvae feed in the productive mudflats. For the remaining tropical habitats, the insect communities have diversity patterns that are largely congruent across guilds. The discovery of such a sizeable and distinct insect fauna in a globally threatened habitat underlines how little is known about global insect biodiversity. We here show how such knowledge gaps can be closed quickly with new cost-effective NGS barcoding techniques.


Modeling the lagged impacts of hourly weather and speed variation factors on the segment crash risk of rural interstate freeways: Applying a space-time-stratified case-crossover design.
In the realm of traditional roadway crash studies, cross-sectional modeling methods have been commonly employed to investigate the intricate relationship between the crash risk of roadway segments and variables including roadway geometrics, weather conditions, and speed distribution. However, these methodologies assume that the explanatory variables and target variable are only associated within the same time period. Although this assumption is well-founded for static factors like roadway geometrics, it proves inadequate when dealing with highly time-varying variables related to weather conditions and speed variation. Recent investigations have unveiled that these time-varying variables may exhibit lagged impacts on segment crash risk, necessitating the adoption of more comprehensive time-series modeling methods. This study employs two interpretable statistical methods, namely the distributed lag model (DLM) and the distributed lag nonlinear model (DLNM), to elucidate meaningful and interpretable patterns of the lagged impacts of weather and speed variation factors on segment crash risk. Empirical evidence based on crash data collected from rural interstate freeways in the state of Texas demonstrates coherent and interpretable lagged impact patterns of these variables. This study's results serve as strong support for the existence of lagged impacts on roadway segment-level crash risk, emphasizing the need for considering time-series effects in future crash modeling research. Furthermore, these findings could offer practical implications for the design of real-time crash warning systems and the effective implementation of variable speed limits to enhance road safety.


Further phenotypic characterization of Kaufman oculocerebrofacial syndrome: report of five new cases and literature review.
Kaufman oculocerebrofacial syndrome is a rare autosomal recessive disorder caused by biallelic variants in UBE3B. Kaufman oculocerebrofacial syndrome is characterized by a recognizable pattern of malformations including moderate to severe intellectual disability, growth deficiency, microcephaly and a distinctive facial gestalt. Common craniofacial features include short upslanting palpebral fissures, blepharophimosis or ptosis, ear anomalies, hearing loss, palate anomalies and stridor/laryngomalacia. The aim of this study was to describe the phenotypic features and the genotype of five new individuals from three unrelated families, and to review systematically the published information of 26 cases. The main features are summarized contributing to further characterize the natural history of the disease. Novel phenotypic features and two novel pathogenic variants in UBE3B are reported: A splice site variant (c.2569-1G > C) and a nonsense variant (c.518C > A, p.Ser173Ter). Kaufman oculocerebrofacial syndrome is likely an underdiagnosed disorder which can be clinically recognized based on its distinctive facial gestalt and relatively homogenous natural history.


Association of thyroid functional disease with mortality in a national cohort of incident hemodialysis patients.
Hypothyroidism is a common condition that disproportionately affects hemodialysis patients. In the general population, hypothyroidism is associated with higher mortality, particularly in populations with underlying cardiovascular risk. Despite their heightened cardiovascular mortality, the impact of hypothyroidism on the survival of hemodialysis patients remains uncertain. To examine whether hypothyroidism is independently associated with higher mortality in hemodialysis patients. Among 8840 incident hemodialysis patients receiving care from a large national dialysis provider from January 2007 to December 2011, we examined the association of hypothyroidism (TSH >5.0 mIU/L) with mortality. Associations between baseline and time-dependent hypothyroidism with all-cause mortality were determined using case-mix adjusted Cox models. In secondary analyses, we examined the impact of low-normal, upper-normal, subclinical range, and overt range TSH levels (TSH ≥ 0.5-3.0, >3.0-5.0, >5.0-10.0, and >10.0 mIU/L, respectively) on mortality risk. The study population consisted of 1928 (22%) hypothyroid and 6912 (78%) euthyroid patients. Baseline and time-dependent hypothyroidism were associated with higher mortality: adjusted hazard ratios (95% confidence intervals) were 1.47 (1.34-1.61) and 1.62 (1.45-1.80), respectively. Compared to low-normal TSH, upper-normal, subclinical hypothyroid, and overt hypothyroid TSH levels were associated with incrementally higher adjusted death risk in baseline and time-dependent analyses. In time-dependent analyses, the hypothyroidism-mortality association was increasingly stronger across higher body mass index strata. Hypothyroidism as well as upper-normal TSH levels are associated with higher mortality in hemodialysis patients. Further studies are needed to determine whether restoration of TSH to low-normal levels with thyroid hormone replacement therapy ameliorates adverse outcomes in hemodialysis patients.


Structure-Interaction Relationship of Polymyxins with Lung Surfactant.
Multidrug-resistant Gram-negative bacteria present an urgent and formidable threat to the global public health. Polymyxins have emerged as a last-resort therapy against these 'superbugs'; however, their efficacy against pulmonary infection is poor. In this study, we integrated chemical biology and molecular dynamics simulations to examine how the alveolar lung surfactant significantly reduces polymyxin antibacterial activity. We discovered that lung surfactant is a phospholipid-based permeability barrier against polymyxins, compromising their efficacy against target bacteria. Next, we unraveled the structure-interaction relationship between polymyxins and lung surfactant, elucidating the thermodynamics that govern the penetration of polymyxins through this critical surfactant layer. Moreover, we developed a novel analog, FADDI-235, which exhibited potent activity against Gram-negative bacteria, both in the presence and absence of lung surfactant. These findings shed new light on the sequestration mechanism of lung surfactant on polymyxins and importantly pave the way for the rational design of new-generation lipopeptide antibiotics to effectively treat Gram-negative bacterial pneumonia.


A mechanistic marker-based screening tool to predict clinical immunogenicity of biologics.
The efficacy and safety of therapeutic proteins are undermined by immunogenicity driven by anti-drug antibodies. Immunogenicity risk assessment is critically necessary during drug development, but current methods lack predictive power and mechanistic insight into antigen uptake and processing leading to immune response. A key mechanistic step in T-cell-dependent immune responses is the migration of mature dendritic cells to T-cell areas of lymphoid compartments, and this phenomenon is most pronounced in the immune response toward subcutaneously delivered proteins. The migratory potential of monocyte-derived dendritic cells is proposed to be a mechanistic marker for immunogenicity screening. Following exposure to therapeutic protein in vitro, dendritic cells are analyzed for changes in activation markers (CD40 and IL-12) in combination with levels of the chemokine receptor CXCR4 to represent migratory potential. Then a transwell assay captures the intensity of dendritic cell migration in the presence of a gradient of therapeutic protein and chemokine ligands. Here, we show that an increased ability of the therapeutic protein to induce dendritic cell migration along a gradient of chemokine CCL21 and CXCL12 predicts higher immunogenic potential. Expression of the chemokine receptor CXCR4 on human monocyte-derived dendritic cells, in combination with activation markers CD40 and IL-12, strongly correlates with clinical anti-drug antibody incidence. Mechanistic understanding of processes driving immunogenicity led to the development of a predictive tool for immunogenicity risk assessment of therapeutic proteins. These predictive markers could be adapted for immunogenicity screening of other biological modalities.


Self-Reported Non-Celiac Wheat Sensitivity in High School Students: Demographic and Clinical Characteristics.
Non-Celiac Wheat Sensitivity (NCWS) has recently been included among the gluten-related disorders. As no biomarkers of this disease exist, its frequency has been estimated based on self-reported symptoms, but to date no data are available about self-reported NCWS in teenagers. To explore the prevalence of self-reported NCWS in a group of high school students and to study their demographic and clinical characteristics. The study was performed between April 2015 and January 2016 in two high schools of a coastal town in the south of Sicily (Italy). A total of 555 students (mean age 17 years, 191 male, 364 female) completed a modified validated questionnaire for self-reported NCWS. The subjects who self-reported NCWS were then compared with all the others. Seven individuals (1.26%) had an established diagnosis of CD. The prevalence of self-reported NCWS was 12.2%, and 2.9% were following a gluten-free diet (GFD). Only 15 out of 68 (23%) NCWS self-reporters had consulted a doctor for this problem and only nine (14%) had undergone serological tests for celiac disease. The NCWS self-reporters very often had IBS symptoms (44%). Self-reported NCWS was found to be common in teenagers, with a frequency of 12.2%; the frequency of GFD use was 2.9%, which was much higher than the percentage of known CD in the same population (1.26%). A greater awareness of the possible implications on the part of the subjects involved, and a more thorough medical approach to the study of self-reported wheat-induced symptoms are required.


Morphological disparity in the cambrian.
An analysis of the range of morphology among arthropods demonstrates that disparity among living arthropods is similar to that in Cambrian arthropods. The range of morphological design resulting from the Cambrian "explosion" has been overestimated, reflecting a tendency to separate as "problematic" taxa that cannot be accommodated in the classification on the basis of the living biota. Problematic taxa are largely an artifact of an inadequate taxonomy. Special evolutionary processes may not be necessary to explain the early radiation of the metazoans.


Long COVID: Rapid Evidence Review.
Postacute sequelae of COVID-19, also known as long COVID, affects approximately 10% to 30% of the hundreds of millions of people who have had acute COVID-19. The Centers for Disease Control and Prevention defines long COVID as the presence of new, returning, or ongoing symptoms associated with acute COVID-19 that persist beyond 28 days. The diagnosis of long COVID can be based on a previous clinical diagnosis of COVID-19 and does not require a prior positive polymerase chain reaction or antigen test result to confirm infection. Patients with long COVID report a broad range of symptoms, including abdominal pain, anosmia, chest pain, cognitive impairment (brain fog), dizziness, dyspnea, fatigue, headache, insomnia, mood changes, palpitations, paresthesias, and postexertional malaise. The presentation is variable, and symptoms can fluctuate or persist and relapse and remit. The diagnostic approach is to differentiate long COVID from acute sequelae of COVID-19, previous comorbidities, unmasking of preexisting health conditions, reinfections, new acute concerns, and complications of prolonged illness, hospitalization, or isolation. Many presenting symptoms of long COVID are commonly seen in a primary care practice, and management can be improved by using established treatment paradigms and supportive care. Although several medications have been suggested for the treatment of fatigue related to long COVID, the evidence for their use is currently lacking. Holistic treatment strategies for long COVID include discussion of pacing and energy conservation; individualized, symptom-guided, phased return to activity programs; maintaining adequate hydration and a healthy diet; and treatment of underlying medical conditions.


Vascular endothelial growth factor (VEGF)-like protein from orf virus NZ2 binds to VEGFR2 and neuropilin-1.
Orf virus, a member of the poxvirus family, produces a pustular dermatitis in sheep, goats, and humans. The lesions induced after infection with orf virus show extensive proliferation of vascular endothelial cells, dilation of blood vessels and dermal swelling. An explanation for the nature of these lesions may lie in the discovery that orf virus encodes an apparent homolog of the mammalian vascular endothelial growth factor (VEGF) family of molecules. These molecules mediate endothelial cell proliferation, vascular permeability, angiogenesis, and lymphangiogenesis via the endothelial cell receptors VEGFR-1 (Flt1), VEGFR-2 (KDR/Flk1), and VEGFR-3 (Flt4). The VEGF-like protein of orf virus strain NZ2 (ORFV2-VEGF) is most closely related in primary structure to VEGF. In this study we examined the biological activities and receptor specificity of the ORFV2-VEGF protein. ORFV2-VEGF was found to be a disulfide-linked homodimer with a subunit of approximately 25 kDa. ORFV2-VEGF showed mitogenic activity on bovine aortic and human microvascular endothelial cells and induced vascular permeability. ORFV2-VEGF was found to bind and induce autophosphorylation of VEGFR-2 and was unable to bind or activate VEGFR-1 and VEGFR-3, but bound the newly identified VEGF165 receptor neuropilin-1. These results indicate that, from a functional viewpoint, ORFV2-VEGF is indeed a member of the VEGF family of molecules, but is unique, however, in that it utilizes only VEGFR-2 and neuropilin-1.


Immunoglobulins on the surface of lymphocytes. II. The bone marrow as the main source of lymphocytes with detectable surface-bound immunoglobulin.
Immunofluorescent studies using live cells from antibody-forming organs and anti-immunoglobulin antibodies demonstrate two populations of small lymphocytes which are differentiated by the presence or absence of Ig on their surface membranes. Most of the lymphocytes with detectable surface Ig appear to derive from cells of the bone marrow, while most of the Ig-negative lymphocytes derive from the thymus. Thus, adult mice thymectomized, lethally irradiated, and transplanted with bone marrow cells showed a normal number of lymphocytes with surface Ig but were depleted of the Ig-negative lymphocytes. Injection of thymocytes into these mice did not result in an increase in the number of lymphocytes with surface Ig in spleen and lymph nodes. Most of the injected thymocytes could be identified by means of histocompatibility markers. Also, the spleen and lymph nodes of neonatally thymectomized mice contained lymphocytes with surface Ig but were depleted of the Ig-negative lymphocytes. Attempts were made to identify light chains on thymocytes by a sensitive radioimmunoassay. In some experiments no light chains were detected and in others a small amount, i.e. no more than 2.6% of the amount present on spleen lymphocytes, could be detected. Whether these low figures are significant or represent a small amount of serum contamination is not clear as yet.


Glatiramer acetate elevates cell production in the mature retinal pigment epithelium.
The retinal-pigmented epithelium (RPE) is critical for visual function. Throughout life, central RPE cells are lost but replenished by peripheral cell production. Glatiramer acetate increases neuronal production in mature brains and is thought to erode age-related deposits in the human retina that are risk factors for macular degeneration. Here, we ask whether this agent also elevates RPE production in mature rat eyes. If so, it may be used to replenish these cells in damaged eyes. Glatiramer acetate was given systemically for 14 days combined with Bromodeoxyuridine (BrdU) to mark cell division. One eye was then processed for the cell cycle marker Ki67 and the other for BrdU. Glatiramer acetate significantly elevated the number of RPE cells in the cell cycle, with more labeled with Ki67. There were also significantly more BrdU-labeled cells over the 14 days, confirming that some cells divided. However, while Ki67 positive cell numbers increased by approximately 100% following examination at one time point, BrdU cell numbers increased by only 3% when averaged per day. Hence, glatiramer acetate induces cells to proliferate, but many may fail either to complete division or to survive. This may have long-term consequences for this tissue.


The effects of semistarvation and parenteral nutrition on the gastric mucosa of rats.
The effects of semistarvation and parenteral nutrition on the gastric mucosa were studied in 24 Wistar rats (250 to 350 grams). The animals were divided into three dietary regimens: Group I-standard rat chow ad libitum; Group II-50 cc. per day of a hyperalimentation solution containing 30% glucose + 5% amino acids; Group III-50 cc. per day of 5% glucose. The animals were fed for a period of 7 days. Gastric mucosal fluxes of Na+, Li+, and H+ then were measured after the gastric instillation of two gastric wash solutions, one primarily an HC1 solution, the other a solution of HC1 plus sodium taurocholate. Gross examination of the gastric mucosal surfaces were recorded. Compared to Group I (oral diet), Groups II and III demonstrated a decrease in volume in gastric secretion during the test period (p less than 0.005); and an increase in net negative hydrogen flux (p less than 0.005). Compared to Group II (hyperalimented), Group III (semistarved) demonstrated an increased net negative H+ flux (p less than 0.01), but no difference in volume of secretion. Only Group III demonstrated a difference in H+ flux after the addition of sodium taurocholate (p less than 0.05). Gastric lesions were significantly increased in Group III, as compared to Groups I and II. Semistarvation renders the gastric mucosa of the rat more susceptible to injury. Adequate intravenous nutrition alone protected against these effects.


Agreement and discrepancy in the evaluation of normal and diabetic oral glucose tolerance test.
Systematic analysis with a five-hour OGTT of 340 subjects representative of people likely to be examined in a center specialized in diabetes detection was performed by multiple discriminant analysis, which provides indices of discrimination for different sets of blood glucose (BG) values. The relative sensitivity and the relative specificity of six different diagnostic methods: Fajans and Conn, Wikerson, WHO, British Diabetic Association, UGDP, and European Study Group of Diabetes Epidemiology were computed, giving a quantitative determination for the degree of discrepancy in the definition of diabetes: only 48 per cent of the subjects are classified in the same way by any of the diagnostic criteria. The time(s) of sampling and the index or indices of OGTT which are the most efficient in screening diabetes were estimated from homogeneous groups of subjects universally recognized as nondiabetic (URND) or as diabetic (URD) according to the different diagnostic methods. Better discriminating power (DP) between URD and URND compared with the maximum DP as measured by D2 of Mahalanobis from the seven BG values of the OGTT is given by the two-hour (70.2 per cent) than by the one-hour (49.5 per cent) BG value when a single value is used; the one-two-hour BG value is the best set of two times (80.7 per cent). The different indices now in use for the classification of the OGTT have been found less effective than the weighted sum of one-two-hour BG values. The difficulty in obtaining highly specific diagnostic tests is discussed in relation to the consequences on a partly automated screening in large populations.


Physician perspectives on incentives to participate in practice-based research: a greater rochester practice-based research network (GR-PBRN) study.
To understand factors associated with primary care physician research participation in a practice-based research network (PBRN) and to compare perspectives by specialty. We surveyed primary care internists, family physicians, and pediatricians in Monroe County, New York, regarding their past experience with research and incentives to participate in practice-based research. We performed descriptive and tabular analyses to assess perceptions and used chi(2) and analysis of variance to compare perceptions across the 3 specialties. The response rate was 33%. The most frequently endorsed aspects of collaboration were the opportunity to enact quality improvement (78%), contribution to clinical knowledge (75%), and intellectual stimulation (65%). Significant differences among the primary care specialties were found in 2 aspects: ((1)) internists were more likely to endorse additional source of income as "important," and family medicine physicians were more likely to cite the opportunity to shape research questions, projects, and journal articles as "important." Physicians across all 3 specialties cited the opportunity to enact quality improvement and contribution to clinical knowledge as important incentives to participating in practice-based research. This supports the importance of strengthening the interface between research and quality improvement in PBRN projects. Further study is needed to assess reasons for differences among specialties if PBRNs are to become successful in research involving adult patients.


Medial and Lateral Plantar Artery Angiosome Rotational Flaps for Transmetatarsal and Lisfranc Amputation in Patients With Compromised Plantar Tissue.
Traditional incision techniques for midfoot amputation might not provide immediate soft tissue coverage of the underlying metatarsal and tarsal bones in the presence of a large plantar soft tissue defect. Patients undergoing transmetatarsal and Lisfranc amputation frequently have compromised plantar tissue in association with neuropathic ulcers, forefoot gangrene, and infection, necessitating wide resection as a part of the amputation procedure. Open amputation will routinely be performed under these circumstances, although secondary healing could be compromised owing to residual bone exposure. Alternatively, the surgeon might elect to perform a more proximal lower extremity amputation, which will allow better soft tissue coverage but compromises function of the lower extremity. A third option for this challenging situation is to modify the plantar flap incision design to incorporate a medial or lateral plantar artery angiosome-based rotational flap, which will provide immediate coverage of the forefoot and midfoot soft tissue defects without excessive shortening of the bone structure. A plantar medial soft tissue defect is treated with the lateral plantar artery angiosome flap, and a plantar lateral defect is treated with the medial plantar artery angiosome flap. Medial and lateral flaps can be combined to cover a central plantar wound defect. Incorporating large rotational flaps requires knowledge of the applicable angiosome anatomy and specific modifications to incision planning and dissection techniques to ensure adequate soft tissue coverage and preservation of the blood supply to the flap. A series of 4 cases with an average follow-up duration of 5.75 years is presented to demonstrate our patient selection criteria, flap design principles, dissection pearls, and surgical staging protocol.


Polygonal deformation of the dural sac in lumbar epidural lipomatosis: anatomic explanation by the presence of meningovertebral ligaments.
In patients with epidural lipomatosis, axial lumbar spine CT and MR images occasionally reveal a geometric, polygonal or stellar, shape of the dural sac. The purpose of this study was to define the anatomic structures responsible for this radiologic appearance. We hypothesized that meningovertebral ligaments could anchor the dura mater to the osteofibrous walls of the spinal canal and account for the geometric deformation of the dural sac. The epidural spaces were examined in 15 adult cadaveric and seven aborted fetal lumbar spines. For macroscopic studies, 70 adult vertebral segments were separated and dissected by removing the extradural fat. For microscopic examination, axial histologic sections were obtained from 35 fetal and five adult undissected vertebral segments. Meningovertebral ligaments were observed in the median, paramedian, and lateral aspects of the anterior and posterior epidural spaces of both adult and fetal lumbar spines. These ligaments anchor the outer surface of the dura mater to the osteofibrous walls of the lumbar canal. They may form an irregular longitudinal septum partitioning the epidural space. Histologic examination demonstrated the fibroelastic composition of these ligaments and suggested their possible perivascular morphogenetic origin. The morphologic and topographic features of the meningovertebral ligaments explain the polygonal, stellar, or Y-shaped deformation of the dural sac observed on axial CT and MR images in patients with lumbar epidural lipomatosis.


Sandy sediment and the bioavailability of 17β-trenbolone to adult female fathead minnows.
Recent studies have detected bioavailable steroids in sediment, however, the mechanism by which these compounds become bioavailable is not completely understood. In this study, two experiments were conducted using a double aquarium system that allowed female fathead minnows to be exposed to sandy sediments without direct contact. In the first experiment, natural sediment from the Elkhorn River (Nebraska, USA) was spiked with 17β-trenbolone. Both the fish in direct contact with the sediment as well as the fish excluded from direct contact experienced significant reductions in the hepatic expression of two estrogen-responsive genes, vitellogenin and estrogen receptor α, indicating molecular defeminization. The natural sediment contained particles ranging in size from sand to clay and it was possible that the fish in experiment 1 were being exposed to trenbolone associated with the very fine particles. The sandy sediment was sieved for experiment 2, and only the particles larger than 250 μm were used. In addition, the experiment was conducted at two different Tb concentrations (1× and 10×). Furthermore nuptial tubercles, a biomarker of exposure to a masculinizing androgen, were also evaluated in the females used in experiment 2. For tubercle number and vtg expression, significant results were obtained from a two-way ANOVA due to Tb concentration, but not tank location or interaction term (location vs. concentration). For ERα expression, results were found in response to Tb concentration and tank location, but not the interaction term. Overall the results from these studies suggest that the primary route of exposure of sediment-associated trenbolone to fish is through ventilation of free compound, rather than ingestion or direct contact with the sediments.


Headache complexity (number of symptom features) differentiates post-traumatic from non-traumatic headaches.
Post-traumatic headaches are a common sequela of mild traumatic brain injury (concussion). It is unclear whether or how these headaches differ phenotypically from primary headaches. Determine whether there is an overarching unobserved latent trait that drives the expression of observed features of post-traumatic headache and other headaches. Data from this post-hoc analysis come from the Warrior Strong Cohort Study conducted from 2010 through 2015. Approximately 25,000 soldiers were screened for concussion history at routine post-deployment health assessments. A random sample was invited to participate, enrolling 1567. Twelve observed headache phenotypic features were used to measure "headache complexity", the latent trait of clinical interest, using single factor confirmatory factor analysis. We compared headache complexity between groups and determined whether headache complexity predicted accessing medical care for headache. Of 1094 soldiers with headaches, 198 were classified as having post-traumatic headache. These headaches were compared to those in the other soldiers (647 without concussion history and 249 with concussion history). Soldiers with post-traumatic headache had greater endorsement of all 12 headache features compared to the soldiers with non-concussive headaches. The confirmatory factor analysis showed good model fit (χ2 (51) = 95.59, p = 0.0002, RMSEA = 0.03, comparative fit index = 0.99, and Tucker-Lewis index = 0.99), providing empirical support for the headache complexity construct. Soldier groups differed in their mean headache complexity level (p < 0.001) such that post-traumatic headache soldiers had greater headache complexity compared to non-concussed soldiers (standardized mean difference = 0.91, 95% confidence interval: 0.72-1.09, p < 0.001 and to concussed soldiers with coincidental headaches standardized mean difference = 0.75, 95% confidence interval: 0.53-0.96, p < 0.001). Increasing headache complexity predicted medical encounters for headache (odds ratio = 1.87, 95% confidence interval: 1.49-2.35, p < 0.001) and migraine (odds ratio = 3.74, 95% confidence interval: 2.33-5.98, p < 0.001) during the year following deployment.Conclusions and relevance: The current study provided support for a single latent trait, characterized by observed headache symptoms, that differentiates between concussive and non-concussive headaches and predicts use of medical care for headache. The single trait confirmatory factor analysis suggests that post-traumatic headaches differ from non-concussive headaches by severity more than kind, based on the symptoms assessed.ClinicalTrials.gov identifier NCT01847040.


Purtscher's retinopathy secondary to pancreatitis. Aspects of the topography of retinal abnormalities.
A 22-year-old man with acute pancreatitis developed sudden bilateral blindness with funduscopic features of Purtscher's retinopathy. The topography of retinal abnormalities has been ascribed to the properties of the retinal capillary network. However, the similarity between the distribution of the retinal abnormalities and the density of the nerve fiber layer suggests that the observed topography may, in fact, result from the anatomical characteristics of the nerve fiber layer.


Dynamic regulation of LFA-1 activation and neutrophil arrest on intercellular adhesion molecule 1 (ICAM-1) in shear flow.
Neutrophil recruitment during acute inflammation is triggered by G-protein-linked chemotactic receptors that in turn activate beta(2) integrin (CD18), deemed a critical step in facilitating cell capture and arrest under the shear force of blood flow. A conformational switch in the I domain allosteric site (IDAS) and in CD18 regulates LFA-1 affinity for endothelial ligands including intercellular adhesion molecule 1 (ICAM-1). We examined the dynamics of CD18 activation in terms of the efficiency of neutrophil capture of ICAM-1, and we correlated this with the membrane topography of 327C, an antibody that recognizes the active conformation of CD18 I-like domain. Adhesion increased in direct proportion to chemotactic stimulus rising 7-fold over a log range of interleukin-8 (IL-8). A threshold dose of approximately 75 pm IL-8, corresponding to ligation of only approximately 10-100 receptors, was sufficient to activate approximately 20,000 CD18 and a rapid boost in the capture efficiency on ICAM-1. This was accompanied by a rapid redistribution of active LFA-1, but not Mac-1, into membrane patches, a necessary component for optimum adhesion efficiency. Shear-resistant arrest on a monolayer of ICAM-1 was reversed within minutes of chemotactic stimulation correlating with a shift from high to low affinity CD18 and dispersal of patches of active CD18. Mobility of active CD18 into high avidity patches was dependent on phosphatidylinositol 3-kinase activity and not F-actin polymerization. The data reveal that the number of chemotactic receptors bound and the topography and lifetime of high affinity LFA-1 tightly regulate the efficiency of neutrophil capture on ICAM-1.


[JAPANESE TRANSLATION AND LINGUISTIC VALIDATION OF THE RECAP OF ATOPIC ECZEMA (RECAP)].
The Recap of atopic eczema (RECAP), a new core outcome of the atopic dermatitis trial, was translated into Japanese and linguistically validated. Translation into Japanese was accomplished according to the ISPOR (International Society for Pharmacoeconomics and Outcome Research) guidelines and the basic guidelines for scale translation. The translation process included two forward translations, reconciliation with native English speakers, third-party back translation, cognitive debriefing, review and harmonization by the original authors. Twenty-seven atopic dermatitis and pediatric specialists from 21 centers in Japan participated in the translation process. Cognitive debriefing was conducted through face-to-face interviews using a think-aloud method with the interview guide including questions about comprehensibility, relevance, comprehensiveness, recall period and suggested improvements, based on the COSMIN methodology. No linguistic or cultural problems were encountered in the translation into Japanese. Cognitive debriefings were conducted with 10 adult patients and 10 parents of pediatric patients. Some minor modifications were made following discussion and approval by the research team and the original authors. The Japanese version of RECAP was considered to be understandable, comprehensive and relevant for adult patients and families of pediatric patients. The Japanese version of the RECAP, which has been validated as linguistically equivalent to the original version, is now available. Further evaluation of the measurement properties is needed in the future.


Strategies used to detect auditory signals in small sets of random maskers.
Detection performance for a masked auditory signal of fixed frequency can be substantially degraded if there is uncertainty about the frequency content of the masker. A quasimolecular psychophysical approach was used to examine response strategies in masker-uncertainty conditions, and to investigate the influence of uncertainty when the number of different masker samples was limited to ten or fewer. The task of the four listeners was to detect a 1000-Hz signal that was presented simultaneously with one of ten ten-tone masker samples. The masker sample was either fixed throughout a block of two-interval forced-choice trials or was randomized across or within trials. The primary results showed that: (1) When the signal level was low and the masker sample differed between the two intervals of a trial, most listeners based their responses more on the presence of specific masker samples than on the signal. (2) The detrimental effect of masker uncertainty was clearly evident when only four maskers were randomly presented, and grew as the size of the masker set was increased from two to ten. (3) The slopes of psychometric functions measured with the same masker samples differed among the fixed and two random-masker conditions. (4) There were large differences in the influence of masker uncertainty across masker samples and listeners. These data demonstrate the great susceptibility of human listeners to the influence of masker uncertainty and the ability of quasimolecular investigations to reveal important aspects of behavior in uncertainty condition.


Demographic, socio-economic, obstetric, and behavioral factors associated with small-and large-for-gestational-age from a prospective, population-based pregnancy cohort in rural Nepal: a secondary data analysis.
In South Asia, a third of babies are born small-for-gestational age (SGA). The risk factors are well described in the literature, but many studies are in high-and-middle income countries or measure SGA on facility births only. There are fewer studies that describe the prevalence of risk factors for large-for-gestational age (LGA) in low-income countries. We aim to describe the factors associated with SGA and LGA in a population-based cohort of pregnant women in rural Nepal. This is a secondary data analysis of community-based trial on neonatal oil massage (22,545 women contributing 39,479 pregnancies). Demographic, socio-economic status (SES), medical/obstetric history, and timing of last menstruation were collected at enrollment. Vital signs, illness symptoms, and antenatal care (ANC) attendance were collected throughout the pregnancy and neonatal weight was measured for live births. We conducted multivariate analysis using multinomial, multilevel logistic regression, reporting the odds ratio (OR) with 95% confidence intervals (CIs). Outcomes were SGA, LGA compared to appropriate-for-gestational age (AGA) and were multiply imputed using birthweight recalibrated to time at delivery. SGA was associated with nulligravida (OR: 2.12 95% CI: 1.93-2.34), gravida/nulliparous (OR: 1.86, 95% CI: 1.26-2.74), interpregnancy intervals less than 18 months (OR: 1.16, 95% CI: 1.07-1.27), and poor appetite/vomiting in the second trimester, (OR: 1.27, 95% CI: 1.19-1.35). Greater wealth (OR: 0.78, 95% CI: 0.69-0.88), swelling of hands/face in the third trimester (OR: 0.81, 95% CI: 0.69-0.94) parity greater than five (OR: 0.77, 95% CI: 0.65-0.92), male fetal sex (OR: 0.91, 95% CI: 0.86-0.98), and increased weight gain (OR: 0.93 per weight kilogram difference between 2nd and 3rd trimester, 95% CI: 0.92-0.95) were protective for SGA. Four or more ANC visits (OR: 0.53, 95% CI: 0.41-0.68) and respiratory symptoms in the third trimester (OR: 0.67, 95% CI: 0.54-0.84) were negatively associated with LGA, and maternal age < 18 years (OR: 1.39, 95% CI: 1.03-1.87) and respiratory symptoms in the second trimester (OR: 1.27, 95% CI: 1.07-1.51) were positively associated with LGA. Our findings are in line with known risk factors for SGA. Because the prevalence and mortality risk of LGA babies is low in this population, it is likely LGA status does not indicate underlaying illness. Improved and equitable access to high quality antenatal care, monitoring for appropriate gestational weight gain and increased monitoring of women with high-risk pregnancies may reduce prevalence and improve outcomes of SGA babies. The study used in this secondary data analysis was registered at Clinicaltrials.gov NCT01177111.