Ingredients, structure and reconstitution properties of instant powder foods and the potential for healthy product development: a comprehensive review.
Instant foods are widely presented in powder forms across different food segments, which potentially can be formulated with functional or beneficial compounds to provide health benefits. Many reconstituted instant powder foods form colloidal suspensions with complex structures. However, designing instant powder food could be challenging due to the structural complexity and high flexibility in formulation. This review proposed a new classification method for instant powder foods according to the solubility of ingredients and the structure of the reconstituted products. Instant powder foods containing insoluble ingredients are discussed. It summarised challenges and current advances in powder treatments, reconstitution improvement, and influences on food texture and structure to facilitate product design in related industries. The characteristics and incorporation of the main ingredients and ingredients with health benefits in product development were reviewed. Different products vary significantly in the ratios of macronutrients. The macronutrients have limited solubility in water. After being reconstituted by water, the insoluble components are dispersed and swell to form colloidal dispersions with complex structures and textures. Soluble components, which dissolve in the continuous phase, may facilitate the dispersing process or influence the solution environment. The structure of reconstituted products and destabilising factors are discussed. Both particle and molecular structuring strategies have been developed to improve wettability and prevent the formation of lumps and, therefore, to improve reconstitution properties. Various types of instant food have been developed based on healthy or functional ingredients and exhibit positive effects on the prevention of non-communicable diseases and overall health. Less processed materials and by-products are often chosen to enhance the contents of dietary fibre and phenolic compounds. The enrichment of phenolic compounds, dietary fibres and/or probiotics tend to be simultaneous in plant-based products. The process of the ingredients and the formulation of products must be tailored to design the desired structure and to improve the reconstitution property.


Anti-HCV, nucleotide inhibitors, repurposing against COVID-19.
A newly emerged Human Coronavirus (HCoV) is reported two months ago in Wuhan, China (COVID-19). Until today >2700 deaths from the 80,000 confirmed cases reported mainly in China and 40 other countries. Human to human transmission is confirmed for COVID-19 by China a month ago. Based on the World Health Organization (WHO) reports, SARS HCoV is responsible for >8000 cases with confirmed 774 deaths. Additionally, MERS HCoV is responsible for 858 deaths out of about 2500 reported cases. The current study aims to test anti-HCV drugs against COVID-19 RNA dependent RNA polymerase (RdRp). In this study, sequence analysis, modeling, and docking are used to build a model for Wuhan COVID-19 RdRp. Additionally, the newly emerged Wuhan HCoV RdRp model is targeted by anti-polymerase drugs, including the approved drugs Sofosbuvir and Ribavirin. The results suggest the effectiveness of Sofosbuvir, IDX-184, Ribavirin, and Remidisvir as potent drugs against the newly emerged HCoV disease. The present study presents a perfect model for COVID-19 RdRp enabling its testing in silico against anti-polymerase drugs. Besides, the study presents some drugs that previously proved its efficiency against the newly emerged viral infection.


Fibrillation of alpha-synuclein triggered by bacterial endotoxin and lipid vesicles is modulated by N-terminal acetylation and familial Parkinson's disease mutations.
It has been hypothesized that Parkinson's disease (PD) may be initiated in the gastrointestinal tract, before manifesting in the central nervous system. In this respect, it was demonstrated that lipopolysaccharide (LPS), an endotoxin from gram-negative bacteria, accelerates the in vitro formation of alpha-synuclein (aSyn) fibrils, whose intracellular deposits is a histological hallmark of the degeneration of dopaminergic neurons in PD. Herein, N-terminal acetylation and missense mutations of aSyn (A30P, A53T, E46K, H50Q and G51D) linked to rare, early-onset forms of familial PD were investigated regarding their effect on aSyn aggregation stimulated by either LPS or small unilamellar lipid vesicles (SUVs). Our findings indicated that LPS as well as SUVs induce the fibrillation of N-terminally acetylated wild-type aSyn (Ac-aSyn-WT) more remarkably than the non-acetylated protein, while the LPS-free protein alone did not undergo fibrillation under our assay conditions. In addition, with the exception of A30P, PD mutations increased the fibrillation of Ac-aSyn in the presence of LPS compared with Ac-aSyn-WT. The most pronounced effect of LPS was noticed for A53T, as observed when either Thioflavin-T or JC-1 were used as fluorescent probes for fibrils. Overall, our results suggest for the first time the existence of a synergy between LPS and PD mutations//N-terminal acetylation toward aSyn fibrillation.


Stress and vascular responses: mitogen-activated protein kinases and activator protein-1 as promising therapeutic targets of vascular remodeling.
Mitogen-activated protein kinases (MAP kinases), including extracellular signal-regulated kinase (ERK), c-Jun NH(2)-terminal kinase (JNK), and p38, play a central role in cellular responses by various stress stimuli such as cell proliferation, apoptosis, migration, or gene expression. Furthermore, activator protein-1 (AP-1), a transcription factor which can be activated by MAP kinases, also is involved in a variety of celllar responses, as well as MAP kinases. MAP kinases and AP-1 are significantly activated in vascular tissues by hypertension, angiotensin II, or balloon injury. We have made dominant negative mutants of MAP kinases or c-Jun, to specifically inhibit in vivo activation of MAP kinases or AP-1. Vascular gene transfer of each dominant negative mutant of MAP kinases or c-Jun prevents intimal hyperplasia after balloon injury, which is associated with the inhibition of smooth muscle cell proliferation in the intima and the media and probably also associated with inhibition of smooth muscle cell migration. However, in vitro findings on cultured vascular smooth muscle cells suggest that the molecular mechanism underlying inhibition of intimal hyperplasia may be different among each dominant negative mutant of MAP kinases and c-Jun. MAP kinases and c-Jun seem to be the promising therapeutic target for vascular remodeling.


Cartilage pressure distributions provide a footprint to define female anterior cruciate ligament injury mechanisms.
Bone bruises located on the lateral femoral condyle and posterolateral tibia are commonly associated with anterior cruciate ligament (ACL) injuries and may contribute to the high risk for knee osteoarthritis after ACL injury. The resultant footprint (location) of a bone bruise after ACL injury provides evidence of the inciting injury mechanism. Purpose/ (1) To analyze tibial and femoral articular cartilage pressure distributions during normal landing and injury simulations, and (2) to evaluate ACL strains for conditions that lead to articular cartilage pressure distributions similar to bone bruise patterns associated with ACL injury. The hypothesis was that combined knee abduction and anterior tibial translation injury simulations would demonstrate peak articular cartilage pressure distributions in the lateral femoral condyle and posterolateral tibia. The corollary hypothesis was that combined knee abduction and anterior tibial translation injury conditions would result in the highest ACL strains. Descriptive laboratory study. Prospective biomechanical data from athletes who subsequently suffered ACL injuries after testing (n = 9) and uninjured teammates (n = 390) were used as baseline input data for finite element model comparisons. Peak articular pressures that occurred on the posterolateral tibia and lateral femoral condyle were demonstrated for injury conditions that had a baseline knee abduction angle of 5°. Combined planar injury conditions of abduction/anterior tibial translation, anterior tibial translation/internal tibial rotation, or anterior tibial translation/external tibial rotation or isolated anterior tibial translation, external tibial rotation, or internal tibial rotation resulted in peak pressures in the posterolateral tibia and lateral femur. The highest ACL strains occurred during the combined abduction/anterior tibial translation condition in the group that had a baseline knee abduction angle of 5°. The results of this study support a valgus collapse as the major ACL injury mechanism that results from tibial abduction rotations combined with anterior tibial translation or external or internal tibial rotations. Reduction of large multiplanar knee motions that include abduction, anterior translation, and internal/external tibial motions may reduce the risk for ACL injuries and associated bone bruises. In particular, prevention of an abduction knee posture during initial contact of the foot with the ground may help prevent ACL injury.


A rational approach toward determining the candidacy of the prostatic cancer patient for cryoimmunotherapy: an interim report.
While receiving only cursory attention compared to that of other malignant neoplasms, studies of patients with prostatic cancer have suggested that host responses, mediated in part by immunobiologic factors, may play a significant role in the pathogenesis and thus, the therapeutic management of this disease. For nearly a decade cryosurgery has gradually attained some degree of acceptance as one of the efficacious treatment modalities available in the therapeutic armamentarium for prostatic cancer. In addition to the localized destruction of the primary tumour, reports of secondary tumour cell destruction, i.e. of distant metastases, have supported the suggestion that cryosurgery, as documented in various animal species, may also be immunopotentiating in man. Although our understanding of immunopotentiation of the host's response to tumour via cryosurgery is far from complete, information derived from experimental studies demonstrating that the developement of an immunologic response following cryosurgery of the prostate may be attributed to androgenically and ontogenically dependent autoantigens may have clinical relevance. This together with knowledge that in the therapeutic management of the patient with prostatic cancer we are confronted with the treatment of an individual who, in the majority of cases, possesses waning immunocompetence, emphasizes the potential significance of evaluating what may be referred to as the 'cryosensitivity' of the prospective cryosurgical patient. At present this 'cryosensitivity' may be said to be dependent upon: (1) concentration of prostatic tissue [secretory(tumour?)]-specific or tumour-associated auto-(neo?) antigen(s); (2) physiologic state (elaboration of androgen), and (3) immunocompetence. It is hopeful that evaluation of these parameters may provide a rational approach toward determining the acceptability of cryosurgery as the treatment of choice in a given patient.


Metabolic rates and the energetic cost of external tag attachment in juvenile blacktip sharks Carcharhinus limbatus.
This study reports on the metabolic rate of the blacktip shark Carcharhinus limbatus and the energetic costs of external tag attachment. Metabolic rates, swimming speed and tail-beat (BT ) frequency were measured in a static respirometer with untagged animals and animals equipped with a small data logger. Tagged sharks showed significantly higher routine oxygen consumption and lower swimming speeds than untagged animals, indicating that tagging significantly affected the swimming efficiency and energetic requirements in these small sharks, and that these effects must be accounted for when interpreting telemetry data from free-ranging individuals.


Spinal Arachnoid Webs: Presentation, Natural History, and Outcomes in 38 Patients.
Spinal arachnoid webs are rarely described bands of thickened arachnoid tissue in the dorsal thoracic spine. Much is unknown regarding their origins, risk factors, natural history, and outcomes. To present the single largest case series, detailing presenting symptoms and outcomes amongst operative and nonoperative patients, to better understand the role of intervention. This retrospective chart review identified 38 patients with arachnoid webs. Patient demographics, radiologic signs, symptoms, and surgical history data were extracted from the electronic medical record. Symptoms were divided by location and character. 28 patients were successfully contacted for follow up outcome surveys. 26 patients (68%) underwent surgical intervention, 12 (32%) were managed non-operatively. 15 (39%) patients had undergone a previous unsuccessful surgery at a different site for their symptoms prior to arachnoid web diagnosis. Commonly presenting symptoms included myelopathy (68%), focal thoracic back pain (68%), lower extremity weakness (45%), numbness and sensory changes (58%), and lower extremity radicular pain (42%), upper extremity weakness (24%), and radicular pain (37%). Focal thoracic pain was associated with thoracic level (P < .02). Myelopathic symptoms were less common in postoperative patients. Postoperative patients described significantly more upper extremity (P < .01) and thoracic (P < .01) numbness and paresthesias. Surveyed nonoperative patients universally described their symptoms as either stable or worsening. Spinal arachnoid webs present with thoracic myelopathy and back pain but can also present with upper extremity symptoms. Surgical intervention stabilizes or improves symptoms and is well received. Nonoperative patients do not spontaneously improve.


Building and Breaking Bonds via a Compact S-Propargyl-Cysteine to Chemically Control Enzymes and Modify Proteins.
Analogous to reversible post-translational protein modifications, the ability to attach and subsequently remove modifications on proteins would be valuable for protein and biological research. Although bioorthogonal functionalities have been developed to conjugate or cleave protein modifications, they are introduced into proteins on separate residues and often with bulky side chains, limiting their use to one type of control and primarily protein surface. Here we achieved dual control on one residue by genetically encoding S-propargyl-cysteine (SprC), which has bioorthogonal alkyne and propargyl groups in a compact structure, permitting usage in protein interior in addition to surface. We demonstrated its incorporation at the dimer interface of glutathione transferase for in vivo crosslinking via thiol-yne click chemistry, and at the active site of human rhinovirus 3C protease for masking and then turning on enzyme activity via Pd-cleavage of SprC into Cys. In addition, we installed biotin onto EGFP via Sonogashira coupling of SprC and then tracelessly removed it via Pd cleavage. SprC is small in size, commercially available, nontoxic, and allows for bond building and breaking on a single residue. Genetically encoded SprC will be valuable for chemically controlling proteins with an essential Cys and for reversible protein modifications.


Analysis of the Clinical Outcome of Arthrographic Steroid Injection for the Treatment of Adhesive Capsulitis.
Idiopathic adhesive capsulitis is a condition of uncertain etiology characterized by pain and decreased shoulder range of motion (ROM) that occurs without a known intrinsic disorder. Many treatments have been advocated, yet the best option remains unclear. The purpose of this study was to determine if arthrographic injection of the shoulder joint with steroid and local anesthetic results in decreased pain and increased shoulder function in a cohort of patients with idiopathic adhesive capsulitis. This is a retrospective case series of patients who were treated with an arthrographic steroid and local anesthetic injection for adhesive capsulitis. The injections were all given by the same musculoskeletal radiologist using image guidance, and the patients were all from the same surgeon's practice. Patients were evaluated with the Shoulder Pain and Disability Questionnaire (SPADI) and Constant Shoulder Score and their shoulder ROM was tested. Descriptive statistics in the form of counts, percentages, means and standard deviations were used, as well as parametric and non-parametric tests. Thirty-three shoulders in 25 patients were examined. The average length of follow-up was 17 months. Sixty-nine percent of the cohort continued to complain of some shoulder pain in the post-injection follow-up clinic, however, their pain had improved from 8.8/10 pre-injection to 2.2/10 post-injection (p=0.01). The average score of the SPADI Pain, SPADI Disability and Constant Score were 36, 31 and 64, respectively. All patients who received an arthrographic injection using steroid and local anesthetic reported improved pain and mobility. In addition, those who had tried other treatment modalities felt that the injection had been the most beneficial.


Cylindrical coordinate based TG-43U1 parameters for dose calculation around elongated brachytherapy sources.
In 2001, the use of cylindrical coordinates was demonstrated to be more suitable than was the use of polar coordinates for accurate computer calculations during treatment planning for 192Ir intravascular brachytherapy sources. In the present work, we investigated the applicability of cylindrical coordinate-based TG-43U1 parameters for dosimetric evaluation and dose calculations for RadioCoil 103Pd sources (RadioMed Corporation, Tyngsboro, MA) 1.0-cm to 6.0-cm long. For brevity, only the results for sources 1.0-cm, 3.0-cm, and 5.0-cm long are presented here. Dosimetric characteristics of RadioCoil 103Pd sources were calculated in liquid water using the Monte Carlo simulation technique. To demonstrate the suitability of this methodology, the Monte Carlo-simulated dose profiles for a RadioCoil 103Pd source 5.0-cm long at radial distances of 0.5 cm, 0.9 cm, and 1.25 cm were compared with calculated data using TG-43U1 parameters in the polar and cylindrical coordinate systems. In addition, we also used a source 1.0-cm long parameterized using cylindrical coordinates to investigate the application of a linear segmented source (LSS) model originally introduced by our group. The results indicate that, for dose calculation around elongated brachytherapy sources, cylindrical coordinate-based TG-43U1 parameters more accurately represent the dose distribution around an elongated source than the polar coordinate-based parameters. In addition, the LSS model, in conjunction with the cylindrical coordinate-based parameters for a source 1.0-cm long, can be used to replicate the dose distribution around any integral source length. This process eliminates the need to collect and enter data for multiple source lengths into treatment planning systems.


Reading Behind the Lines: The Factors Affecting the Text Reception Threshold in Hearing Aid Users.
The visual Text Reception Threshold (TRT) test (Zekveld et al., 2007) has been designed to assess modality-general factors relevant for speech perception in noise. In the last decade, the test has been adopted in audiology labs worldwide. The 1st aim of this study was to examine which factors best predict interindividual differences in the TRT. Second, we aimed to assess the relationships between the TRT and the speech reception thresholds (SRTs) estimated in various conditions. First, we reviewed studies reporting relationships between the TRT and the auditory and/or cognitive factors and formulated specific hypotheses regarding the TRT predictors. These hypotheses were tested using a prediction model applied to a rich data set of 180 hearing aid users. In separate association models, we tested the relationships between the TRT and the various SRTs and subjective hearing difficulties, while taking into account potential confounding variables. The results of the prediction model indicate that the TRT is predicted by the ability to fill in missing words in incomplete sentences, by lexical access speed, and by working memory capacity. Furthermore, in line with previous studies, a moderate association between higher age, poorer pure-tone hearing acuity, and poorer TRTs was observed. Better TRTs were associated with better SRTs for the correct perception of 50% of Hagerman matrix sentences in a 4-talker babble, as well as with better subjective ratings of speech perception. Age and pure-tone hearing thresholds significantly confounded these associations. The associations of the TRT with SRTs estimated in other conditions and with subjective qualities of hearing were not statistically significant when adjusting for age and pure-tone average. We conclude that the abilities tapped into by the TRT test include processes relevant for speeded lexical decision making when completing partly masked sentences and that these processes require working memory capacity. Furthermore, the TRT is associated with the SRT of hearing aid users as estimated in a challenging condition that includes informational masking and with experienced difficulties with speech perception in daily-life conditions. The current results underline the value of using the TRT test in studies involving speech perception and aid in the interpretation of findings acquired using the test.


Interleukin-1 beta is involved in the pressor response of amygdaloid neurons excited by sodium glutamate.
The effects of interleukin-1beta in the paraventricular nucleus or caudal arcuate nucleus on hypertensive and tachycardiac responses induced by excitation of the central amygdaloid nucleus were studied. We observed that microinjection of sodium glutamate into central amygdaloid nucleus resulted in hypertension and tachycardia. Microinjection of interleukin-1beta into paraventricular nucleus or caudal arcuate nucleus induced significant pressor and tachycardiac responses while pretreatment with microinjection of rabbit anti-rat interleukin-1beta antibody into bilateral paraventricular nuclei or arcuate nuclei attenuated the hypertensive response induced by microinjection of 40 nmol sodium glutamate into central amygdaloid nucleus. The above results suggest that the pressor effect of central amygdaloid nucleus is mediated by interleukin-1beta in paraventricular nucleus or arcuate nucleus.


Intragranular vesiculotubular compartments are involved in piecemeal degranulation by activated human eosinophils.
Eosinophils, leukocytes involved in allergic, inflammatory and immunoregulatory responses, have a distinct capacity to rapidly secrete preformed granule-stored proteins through piecemeal degranulation (PMD), a secretion process based on vesicular transport of proteins from within granules for extracellular release. Eosinophil-specific granules contain cytokines and cationic proteins, such as major basic protein (MBP). We evaluated structural mechanisms responsible for mobilizing proteins from within eosinophil granules. Human eosinophils stimulated for 30-60 min with eotaxin, regulated on activation, normal, T-cell expressed and secreted (RANTES) or platelet activating factor exhibited ultrastructural features of PMD (e.g. losses of granule contents) and extensive vesiculotubular networks within emptying granules. Brefeldin A inhibited granule emptying and collapsed intragranular vesiculotubular networks. By immunonanogold ultrastructural labelings, CD63, a tetraspanin membrane protein, was localized within granules and on vesicles outside of granules, and mobilization of MBP into vesicles within and extending from granules was demonstrated. Electron tomography with three dimension reconstructions revealed granule internal membranes to constitute an elaborate tubular network able to sequester and relocate granule products upon stimulation. We provide new insights into PMD and identify eosinophil specific granules as organelles whose internal tubulovesicular networks are important for the capacity of eosinophils to secrete, by vesicular transport, their content of preformed and granule-stored cytokines and cationic proteins.


Adoption of Moringa oleifera to combat under-nutrition viewed through the lens of the "Diffusion of innovations" theory.
Moringa oleifera, an edible tree found worldwide in the dry tropics, is increasingly being used for nutritional supplementation. Its nutrient-dense leaves are high in protein quality, leading to its widespread use by doctors, healers, nutritionists and community leaders, to treat under-nutrition and a variety of illnesses. Despite the fact that no rigorous clinical trial has tested its efficacy for treating under-nutrition, the adoption of M. oleifera continues to increase. The "Diffusion of innovations theory" describes well, the evidence for growth and adoption of dietary M. oleifera leaves, and it highlights the need for a scientific consensus on the nutritional benefits.


The disulphide bridges in a cellobiohydrolase and an endoglucanase from Trichoderma reesei.
The positions of the disulphide bridges of the 1,4-beta-glucan cellobiohydrolase (CBH I) of the fungus Trichoderma reesei have been investigated. The results can be summarized as follows. (1) The enzyme contains 12 disulphide bridges and no free cysteine residues. (2) The location of six disulphide bridges have been determined experimentally. (3) The bonding patterns of the two disulphide bridges in the C-terminal region is suggested on the basis of internal homology. (4) The remaining four disulphide bridges are put into two groups, each containing four half-cystine residues where two are adjacent. (5) A repeating bonding pattern is observed along the peptide chain and a non-local disulphide bond with an unusually long separation distance links the N-terminal and the C-terminal region. (6) The disulphide-bonded CNBr peptides of a 1,4-beta-glucan glucanohydrolase (endoglucanase II) from T. reesei have been isolated and a disulphide bonding pattern is suggested on the basis of the sequence homology between the two enzymes.


Identification of three transforming proteins encoded by cottontail rabbit papillomavirus.
Cottontail rabbit papillomavirus (CRPV) provides an animal model for human papillomaviruses associated with a high risk of cancer development. So far, nothing is known about the transforming functions of CRPV genes because of the lack of an assay system. We have recently developed two systems to assay for CRPV transforming functions. One is based on the finding that transformation of NIH 3T3 cells by CRPV is considerably increased by deleting sequences in open reading frame L2. The second one is based on the use of a cottontail rabbit skin epithelial cell line, sf1Ep (C. Meyers and F. O. Wettstein, Virology 181:637-646, 1991). Mutations were introduced which abolished expression of the full-length E6 protein (LE6), the short E6 protein (SE6) initiated at the second ATG of E6, the E7 protein, or the E5 protein. Mutations affecting LE6 or E7, but not SE6, reduced transformation of NIH 3T3 and sf1Ep cells. Transformed NIH 3T3 cell lines with mutations in LE6 and E7 did not grow in soft agar, while those with mutations in SE6 and E5 grew with a reduced efficiency. The cell lines with mutations in LE6, SE6, or E7 still did induce tumors in nude mice. These mutations, however, abolished the ability to induce papillomas in rabbits. When expressed individually with a retroviral vector, LE6, SE6, or E7, but not E5, conferred anchorage-independent growth. The level of viral protein expression in these cell lines was generally low, and a comparison of the abundance of virus-specific mRNA showed that cell lines contained 20 to 50 times less mRNA than a cottontail rabbit papilloma. These data demonstrate that CRPV encodes at least three transforming proteins.


Stakeholder views of rural community-based medical education: a narrative review of the international literature.
Rural community-based medical education (RCBME), in which medical student learning activities take place within a rural community, requires students, clinical teachers, patients, community members and representatives of health and government sectors to actively contribute to the educational process. Therefore, academics seeking to develop RCBME need to understand the rural context, and the views and needs of local stakeholders. The aim of this review is to examine stakeholder experiences of RCBME programmes internationally. This narrative literature review of original research articles published after 1970 utilises Worley's symbiosis model of medical education as an analysis framework. This model proposes that students experience RCBME through their intersection with multiple clinical, social and institutional relationships. This model seeks to provide a framework for considering the intersecting relationships in which RCBME programmes are situated. Thirty RCBME programmes are described in 52 articles, representing a wide range of rural clinical placements. One-year longitudinal integrated clerkships for penultimate-year students in Anglosphere countries were most common. Such RCBME enables students to engage in work-integrated learning in a feasible manner that is acceptable to many rural clinicians and patients. Academic results are not compromised, and a few papers demonstrate quality improvement for rural health services engaged in RCBME. These programmes have delivered some rural medical workforce outcomes to communities and governments. Medical students also provide social capital to rural communities. However, these programmes have significant financial cost and risk student social and educational isolation. Rural community-based medical education programmes are seen as academically acceptable and can facilitate symbiotic relationships among students, rural clinicians, patients and community stakeholders. These relationships can influence students' clinical competency and professional identity, increase graduates' interest in rural careers, and potentially improve rural health service stability. Formal prospective stakeholder consultations should be published in the literature.


Development of a prolonged calcium plateau in hippocampal neurons in rats surviving status epilepticus induced by the organophosphate diisopropylfluorophosphate.
Organophosphate (OP) compounds are among the most lethal chemical weapons ever developed and are irreversible inhibitors of acetylcholinesterase. Exposure to majority of OP produces status epilepticus (SE) and severe cholinergic symptoms that if left untreated are fatal. Survivors of OP intoxication often suffer from irreversible brain damage and chronic neurological disorders. Although pilocarpine has been used to model SE following OP exposure, there is a need to establish a SE model that uses an OP compound in order to realistically mimic both acute and long-term effects of nerve agent intoxication. Here we describe the development of a rat model of OP-induced SE using diisopropylfluorophosphate (DFP). The mortality, behavioral manifestations, and electroencephalogram (EEG) profile for DFP-induced SE (4 mg/kg, sc) were identical to those reported for nerve agents. However, significantly higher survival rates were achieved with an improved dose regimen of DFP and treatment with pralidoxime chloride (25 mg/kg, im), atropine (2 mg/kg, ip), and diazepam (5 mg/kg, ip) making this model ideal to study chronic effects of OP exposure. Further, DFP treatment produced N-methyl-D-aspartate (NMDA) receptor-mediated significant elevation in hippocampal neuronal [Ca(2+)](i) that lasted for weeks after the initial SE. These results provided direct evidence that DFP-induced SE altered Ca(2+) dynamics that could underlie some of the long-term plasticity changes associated with OP toxicity. This model is ideally suited to test effective countermeasures for OP exposure and study molecular mechanisms underlying neurological disorders following OP intoxication.


Survival of Patients on Hemodialysis with Erectile Dysfunction.
Background and objectives: In patients on hemodialysis, erectile dysfunction is an independent mortality factor. This study aimed to determine the risk factors that affect the survival of hemodialysis patients with erectile dysfunction. Materials and Methods: During a seven-year period, erectile dysfunction was identified among the fatalities reported in patients receiving chronic hemodialysis, on the basis of the International Index of Erectile Function questionnaire. The study covered 70 patients of mean age 57 ± 6.7 years. During the examined period, 42 (60%) patients died at the mean age 57 ± 6.8 years. The study was completed by 28 (40%) patients, aged 57 ± 6.55 years. Laboratory, demographic, anthropometric, and clinical characteristics were recorded using standard methods. Results: Statistically significant differences between the two groups of respondents were found concerning dialysis duration (p < 0.001), number of leukocytes (p = 0.003), adequacy of hemodialysis (p = 0.004), intima media thickness of the carotid artery (p < 0.001), presence of cardiovascular disease (p = 0.03), residual diuresis (p = 0.04), and hemodiafiltration (p < 0.001). Hemodialysis adequacy (B = -9.634; p = 0.017), intima media thickness (B = 0.022; p = 0.003), residual diuresis (B = -0.060; p = 0.007), and lower rates of cardiovascular disease (B = 0.176; p = 0.034) were significant survival predictors among our patients with erectile dysfunction. Conclusions: Risk factors that are associated with improved survival of patients on hemodialysis with erectile dysfunction in our study are: preserved diuresis, high-quality hemodialysis, lower incidence of cardiovascular disease, and less thickening of the intima media of the carotid arteries.


Diverse calcium channel types are present in the human placental syncytiotrophoblast basal membrane.
The functional expression of calcium channels has been scarcely studied in human placental syncytiotrophoblast. We have presently sought to characterize Ca(2+) currents of the healthy syncytiotrophoblast basal membrane using purified basal membranes reconstituted in giant liposomes subjected to patch-clamp recordings. We detected presence of channels with high permeability to Ca(2+) (relative PCa/PK up to 99.5) using K(+) solutions in symmetric conditions. Recordings performed in Ba(2+) gradients showed Ba(2+)-conducting channels in 100% of experiments. Ba(2+) total patch currents were consistently blocked by addition of NiCl(2), Nifedipine (L-type voltage-gated calcium channel blocker) or Ruthenium Red (TRPV5-TRPV6 channel blocker); Nifedipine and Ruthenium Red exerted a synergic blocking effect on Ba(2+) total patch currents. Immunohistochemistry of placental villi sections evidenced presence of alpha(1) subunit of voltage-gated calcium channels and TRPV5-TRPV6 channels in basal and apical syncytiotrophoblast plasma membranes; these three calcium channels were also detected in purified basal and apical fractions using Western blot. These results show the presence of three types of calcium channels in the syncytiotrophoblast basal membrane by both functional and molecular means. These basal membrane calcium channels would not be directly involved in mother-to-fetus Ca(2+) transport, but could participate in other relevant trophoblast processes, such as exocytosis and Ca(2+) transport regulation.


Sclerosing stromal tumor of the ovary: a clinicopathologic, immunohistochemical, ultrastructural, and cytogenetic analysis with special reference to its vasculature.
Sclerosing stromal tumor (SST) is a rare ovarian neoplasm occurring predominantly in young women and is histologically characterized by cellular heterogeneity, prominent vasculature, and a pseudolobular appearance composed of cellular and hypocellular areas. In the current study, three cases of SST were ultrastructurally examined and the tumors were found to be composed of several kinds of cells, i.e., luteinized thecalike cells, spindle-shaped fibroblastic cells, and primitive mesenchymal cells. These findings thus supported the ovarian stromal origin of SST. Twelve cases of SST also were analyzed immunohistochemically and demonstrated an expression of vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) in the luteinized thecalike cells and its receptor, fms-like tyrosine kinase 1 (flt-1), in capillary to medium-sized blood vessels. Reverse transcription-polymerase chain reaction (RT-PCR) also showed an expression of VPF/VEGF messenger RNA in SSTs. Accordingly, the characteristic vasculature and edema of SSTs were considered to be associated with the expression of VPF/VEGF. In addition, a fluorescence in situ hybridization (FISH) analysis also showed cells with three copy number of chromosome 12 in 13-21% of all examined SST cells, which suggested the presence of chromosome 12 trisomy in SSTs as well as in other ovarian stromal tumors.


[Familial partial epilepsies].
To investigate the clinical and genetic characteristics of familial partial epilepsies. Family history of seizures was questioned in all patients followed in our epilepsy clinics, from October 1997 to December 1998. Those with positive family history were further investigated and detailed pedigrees were obtained. All possibly affected individuals available underwent clinical evaluation. Seizures and epilepsy syndromes were classified according to the ILAE recommendations. Whenever possible, EEG and MRI were performed. Positive family history was identified in 32 unrelated patients. A total of 213 possibly affected individuals were identified, 161 of whom have been evaluated. The number of affected subjects per family ranged from two to 23. Temporal lobe epilepsy (TLE) was identified in 22 families (68%), frontal lobe epilepsy in one family (3%), partial epilepsy with centrotemporal spikes in five families (15%), and other benign partial epilepsies of childhood in four families (12%). Most of the affected individuals in the TLE families (69%) had clinical and/or EEG characteristics of typical TLE. However, the severity of epilepsy was variable, with 76% of patients with spontaneous seizure remission or good control with medication and 24% with refractory seizures, including 7 patients that underwent surgical treatment. In the other 10 families, we identified 39 possibly affected subjects, 23 of whom were evaluated. All had good seizure control (with or without medication) except for one patient with frontal lobe epilepsy. Pedigree analysis suggested autosomal dominant inheritance with incomplete penetrance in all families. Family history of seizures is frequent among patients with partial epilepsies. The majority of our families had TLE and its expression was not different from that observed in sporadic cases. The identification of genes involved in partial epilepsies may be usefull in classification of syndromes, to stablish prognosis and optimal treatment.


Increased Plasma Levels of Chemoresistance-Inducing Fatty Acid 16:4(n-3) After Consumption of Fish and Fish Oil.
Our research group previously identified specific endogenous platinum-induced fatty acids (PIFAs) that, in picomolar quantities, activate splenic macrophages leading to resistance to chemotherapy in mouse models. Fish oil was shown to contain the PIFA 16:4(n-3) (hexadeca-4,7,10,13-tetraenoic acid) and when administered to mice neutralized chemotherapy activity. Because patients with cancer frequently use fish oil supplements, we set out to determine exposure to 16:4(n-3) after intake of fish or fish oil. (1) In November 2011, 400 patients with cancer undergoing treatment at the University Medical Center Utrecht were surveyed to determine their use of fish oil supplements; 118 patients responded to the questionnaire (30%); (2) pharmacokinetic analysis of the 16:4(n-3) content of 6 fish oils and 4 fishes was carried out; (3) from April through November 2012, a healthy volunteer study was performed to determine 16:4(n-3) plasma levels after intake of 3 different brands of fish oil or 4 different fish species. Thirty healthy volunteers were randomly selected for the fish oil study; 20 were randomly selected for the fish study. These studies were supported by preclinical tumor experiments in mice to determine chemoresistance conducted between September 2011 and December 2012. (1) Rate of use of fish oil supplements among patients undergoing cancer treatment at our institution; (2) levels of 16:4(n-3) present in 3 brands of fish oil and 4 species of fish; and (3) plasma levels of 16:4(n-3) present in healthy volunteers after consuming fish oil or fish. Eleven percent of respondents reported using omega-3 supplements. All fish oils tested contained relevant amounts of 16:4(n-3), from 0.2 to 5.7 µM. Mouse experiments showed that addition of 1 µL of fish oil to cisplatin was sufficient to induce chemoresistance, treatment having no impact on the growth rate of tumors compared with vehicle-treated controls (estimated tumor volume difference, 44.1 mm3; P > .99). When the recommended daily amount of 10 mL of fish oil was administered to healthy volunteers, rises in plasma 16:4(n-3) levels were observed, reaching up to 20 times the baseline levels. Herring and mackerel contained high levels of 16:4(n-3) in contrast to salmon and tuna. Consumption of fish with high levels of 16:4(n-3) also resulted in elevated plasma levels of 16:4(n-3). All tested fish oils and herring and mackerel fishes contained relevant levels of fatty acid 16:4(n-3), a fatty acid with chemotherapy-negating effects in preclinical models. After ingestion of these fish oils or fishes, 16:4(n-3) was rapidly taken up in the plasma of human volunteers. Until further data become available, fish oil and fish containing high levels of 16:4(n-3) may best be avoided on the days surrounding chemotherapy.


Temporal and contextual consistency of leadership in homing pigeon flocks.
Organized flight of homing pigeons (Columba livia) was previously shown to rely on simple leadership rules between flock mates, yet the stability of this social structuring over time and across different contexts remains unclear. We quantified the repeatability of leadership-based flock structures within a flight and across multiple flights conducted with the same animals. We compared two contexts of flock composition: flocks of birds of the same age and flight experience; and, flocks of birds of different ages and flight experience. All flocks displayed consistent leadership-based structures over time, showing that individuals have stable roles in the navigational decisions of the flock. However, flocks of balanced age and flight experience exhibited reduced leadership stability, indicating that these factors promote flock structuring. Our study empirically demonstrates that leadership and followership are consistent behaviours in homing pigeon flocks, but such consistency is affected by the heterogeneity of individual flight experiences and/or age. Similar evidence from other species suggests leadership as an important mechanism for coordinated motion in small groups of animals with strong social bonds.


Lack of efficacy of light reduction in preventing retinopathy of prematurity. Light Reduction in Retinopathy of Prematurity (LIGHT-ROP) Cooperative Group.
Hospital-nursery lighting has been suggested as a factor in causing retinopathy of prematurity. Despite ongoing debate, a causal relation has not been established. We conducted a prospective, randomized, multicenter study of the effects of light reduction on 409 premature infants with birth weights of less than 1251 g and gestational ages of less than 31 weeks. Two hundred five infants were exposed to reduced light, and 204 to typical nursery lighting. The amount of light reaching the infants' eyes was reduced within 24 hours after birth by placing goggles on the infants that reduced visible-light exposure by 97 percent and ultraviolet-light exposure by 100 percent. The babies wore the goggles until 31 weeks' postconceptional age or 4 weeks after birth, whichever was longer. Once the goggles were removed, ophthalmologists masked to the treatment assignments assessed the infants for retinopathy of prematurity at least biweekly for up to 13 weeks. There were 188 infants in the group that wore goggles and 173 in the control group who survived and were available for follow-up. The mean birth weights were 906 g in the goggles group and 914 g in the control group; the mean gestational ages were 27.4 weeks and 27.2 weeks, respectively. The mean ambient-light level adjacent to the infants' faces was 399 lux for the goggles group and 447 lux for the control group. Retinopathy of prematurity was diagnosed in 102 infants (54 percent) in the goggles group and 100 (58 percent) in the control group (relative risk, 0.9; 95 percent confidence interval, 0.8 to 1.1; P=0.50). A reduction in ambient-light exposure does not alter the incidence of retinopathy of prematurity.


Can black tea influence plasma total homocysteine concentrations?
Polyphenols can act as acceptors of methyl groups during the metabolism of methionine to homocysteine. This may result in elevations in plasma total homocysteine (tHcy) concentrations after ingestion of polyphenol-rich beverages such as tea. Our major objective was to determine whether regular, moderate-to-high intakes of black tea alter tHcy concentrations. We also assessed the relation between the degree of O-methylation of tea-derived polyphenols and the change in tHcy with regular ingestion of tea. Twenty-two subjects completed a randomized, controlled crossover study. Subjects consumed 1250 mL black tea/d (5 cups each containing 2 g tea leaves in 250 mL boiled water) and 1250 mL hot water/d for 4 wk each. Fasting tHcy concentrations and 24-h urinary excretion of 4-O-methylgallic acid (4OMGA, the major O-methylated metabolite of gallic acid) were measured at the end of each period. 4OMGA was used as a marker of overall O-methylation of tea-derived polyphenols. Black tea did not significantly alter mean (+/- SEM) tHcy concentrations (9.9 +/- 0.5 and 10.0 +/- 0.5 micro mol/L for the hot water and black tea periods, respectively). However, the increased excretion of 4OMGA as a consequence of black tea consumption was positively associated with the change in tHcy from the hot water period to the black tea period (r = 0.55, P = 0.008). Subjects in the bottom quartile of increase in 4OMGA excretion had a significant decrease in tHcy (-0.28 +/- 0.10 micro mol/L; P = 0.046), and those in the top quartile had a significant increase in tHcy (0.78 +/- 0.16 micro mol/L; P = 0.005). Overall, regular ingestion of black tea did not alter mean tHcy concentrations. However, individual differences in O-methylation of polyphenolic compounds may influence the ultimate effects of black tea on tHcy.


The interplay between trait anxiety, cognitive biases and attentional control in healthy individuals with psychotic-like experiences.
Although both anxiety and cognitive biases contribute to psychosis, it is still unclear whether these factors interact in their influence on psychotic symptoms. The aim of the study was to examine the interactions between trait anxiety, cognitive biases and delusion-like experiences (PLEs) in a non-clinical sample; the moderational role of attentional control on this relationship was also considered in the study. Two subgroups of participants, 92 individuals (M=24.76; SD=6.33) with heightened (HA) trait anxiety and 78 individuals (M=23.09; SD=5.66) with lowered (LA) trait anxiety took part in the study. Anxiety, cognitive biases and attentional control were measured using self-report questionnaires. Regression analyses and moderation analyses were performed. External attribution bias predicted psychotic-like experiences in both groups, whereas attention to threat bias and belief inflexibility predicted PLEs within LA group. Further moderation analyses revealed that in LA group attention to threat bias increases PLEs only among individuals with a high and moderate ability to focus attention. The results indicated that trait anxiety is an important factor influencing the relationship between cognitive biases and PLEs, and that the ability to voluntarily focus attention is a significant moderator of the linkage between attention to threat bias and delusion-like experiences among low-anxious healthy individuals.


Closed-loop subcutaneous insulin infusion algorithm with a short-acting insulin analog for long-term clinical application of a wearable artificial endocrine pancreas.
Considering the management and safety of the insulin delivery route when a wearable artificial endocrine pancreas is applied to ambulatory diabetic patients on a long-term basis, we developed a s.c. insulin infusion algorithm by analyzing the dynamics of a s.c. injected short-acting insulin analog (Insulin Lispro) by a three-compartment model. Principally the insulin infusion algorithm was developed as a transfer function with the first-order delay in both proportional and derivative actions to blood glucose concentrations. The parameters for this algorithm were calculated to simulate a physiological plasma insulin profile as closely as possible. By applying this algorithm with regular insulin, diabetic patients showed a 2 h postprandial hyperglycemia and a delayed hyperinsulinemia, followed by hypoglycemic episodes 4-5 h after oral glucose load, just as observed in the computer simulation study. However, using Insulin Lispro, a near-physiological glycemic control (postprandial blood glucose of 153.1 +/- 8.3 mg/100 ml at 60 min and 90.3 +/- 7.1 mg/100 ml at 180 min, respectively) could be achieved without showing any delayed hyperinsulinemia or hypoglycemia. Daily glycemic excursions were also controlled near-physiologically and although the daily insulin requirement (731.7 +/- 160.5 mU/kg/day) was slightly higher, it was not significantly different from that with i.v. insulin infusion (622.3 +/- 142.6 mU/kg/day). These results indicate that the application of s.c. insulin infusion algorithm with Insulin Lispro is feasible for long-term glycemic control with a wearable artificial endocrine pancreas in ambulatory diabetic patients.


[Endogenous interaction of neurotransmitters and aging: studies on the basal ganglia of awake rats].
The interaction of glutamate and dopamine in the basal ganglia of the awake rat during the normal process of aging is revised. Using a novel approach, that of blocking the reuptake of glutamate, the effects of increasing concentrations of endogenous glutamate on the extracellular concentrations of dopamine in striatum and nucleus accumbens in the young rat were investigated. It was found that increasing concentrations of glutamate correlated significantly with increasing concentrations of dopamine in striatum and nucleus accumbens. Moreover the increase of dopamine in both structures was significantly reduced after blockade of NMDA and AMPA/kainate glutamate receptors, suggesting that the increase of dopamine was mediated by glutamate. The interaction glutamate/dopamine expressed by its ratio showed a significant age-related decrease in nucleus accumbens but not in neostriatum, so that to a given amount of glutamate less increase of dopamine is produced. It is suggested that the interaction glutamate-dopamine represents a balanced input to the GABA neuron in the basal ganglia and that during aging this balance is disrupted. It is speculated on the significance of this glutamate-dopamine disruption in relation to the changes in motor behavior found with age.


The number of lymph nodes involved with metastatic disease does not affect outcome in melanoma patients as long as all disease is confined to the sentinel lymph node.
In melanoma, a direct relationship exists between the number of nodes involved with metastatic disease and prognosis. This study was undertaken to determine whether an individual with metastatic disease confined to the sentinel lymph nodes (SLNs) would have a better prognosis than individuals with metastatic disease that has spread to the non-SLNs, regardless of the number of nodes involved. The study group consists of 229 melanoma patients with a positive SLN who underwent regional nodal dissection. Cox proportional hazard regression models were used to assess association of the number of SLNs and non-SLNs involved with disease with overall survival (OS) and disease-free survival (DFS). DFS and OS were unchanged regardless of how many SLNs were positive, as long as all disease was confined to SLNs. Among 183 patients without involvement of non-SLNs, OS remained the same despite an increasing number of SLNs involved (P = .59). This was true after controlling for ulceration, Breslow depth, age, sex, and adjuvant treatment. Once disease was present beyond the SLN, DFS and OS were negatively affected. Among patients with involvement of non-SLNs, there was no statistically significant association between the number of positive SLNs and survival. The risk of mortality increased with the number of non-SLNs involved with metastatic disease (P < .001). The number of regional nodes involved with metastatic disease does not affect DFS and OS if disease is confined to the SLNs. Consideration should be given to specifying SLN versus non-SLN involvement in the American Joint Committee on Cancer staging manual.


Blood transfusion administration in seriously ill patients: an evaluation of SUPPORT data. Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments.
Administration of blood transfusion in seriously ill patients is highly variable. Limited data are available to guide transfusion decisions. To explore characteristics of patients who received blood transfusions and decisions to forego transfusions in the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT). Prospective study of preferences, decision-making, and outcomes. Five teaching hospitals. 9105 patients aged 18 years and older meeting defined diagnostic and illness severity criteria. Data included blood transfusions, demographic characteristics, diagnoses, comorbid conditions, acute physiology score (APS), nutritional support, and functional status before hospitalization. A total of 2863 patients (31.4%) received blood transfusions, usually early in their hospitalization. Transfused patients were more likely male (57.3 %; P = .008), with a younger mean age (56 vs 64 years), significantly higher APS (P < .001), and significantly lower 2- and 6-month survival predictions (P < .001). The patients with acute respiratory failure or multiorgan system failure with sepsis (1714; 59.9% of all patients receiving transfusions), multiorgan system failure with malignancy (480, 16.8%), and cirrhosis (248, 8.7%) were more likely to receive blood than those with other diseases. Few patients made a decision not to receive blood before (5; 0.05%) or after (126; 1.4%) study entry. Most patients with decisions to forego transfusions also had decisions against trying resuscitation. Most transfusions occurred in patients with acute respiratory or multiorgan system failure. Few patients decide to forego transfusions. Additional investigation is necessary to evaluate blood transfusion practices in seriously ill patients.


Short-term improvement of masticatory function after implant restoration.
Dental implants present several advantages over other tooth replacement options. However, there has been little research on masticatory function in relation to implant treatment. Therefore, the aim of the present study was to evaluate the improvement of masticatory function two weeks after implant restoration. Masticatory ability was evaluated with the subjective food intake ability (FIA) and objective mixing ability index (MAI) methods. Fifty-four subjects with first and second missing molars completed the study. The subjects were asked to complete a self-reported questionnaire about 30 different food items, and to chew wax samples 10 times both before and two weeks after implant restoration. A total of 108 waxes were analyzed with an image analysis program. Dental implant restoration for lost molar teeth on one side increased the FIA score by 9.0% (P<0.0001). The MAI score also increased, by 14.3% after implant restoration (P<0.0001). Comparison between the good and poor mastication groups, which were subdivided based on the median MAI score before implant restoration, showed that the FIA score of the poor group was enhanced 1.1-fold while its MAI score was enhanced 2.0-fold two weeks after an implant surgery. Using the FIA and MAI assessment methods, this study showed that masticatory function was improved two weeks after implant restoration. In particular, the enhancement of masticatory function by implant restoration was greater in patients with relatively poor initial mastication than in those with good initial mastication.


An in vitro autoradiographic analysis of mu and delta opioid binding in the hippocampal formation of kindled rats.
Recent studies have shown that opioid peptide levels are altered in hippocampal formation of kindled animals. We therefore studied the distributions of mu and delta opioid binding sites in hippocampal formation of kindled and control rats using quantitative in vitro autoradiography. Animals received daily stimulations of the amygdala until they experienced 3 class 5 seizures. Paired control animals underwent implantation of electrodes but were not stimulated. Mu binding sites were labeled with 125I-FK-33824. Twenty-four hours after the last kindled seizure, mu binding was decreased by 32% in stratum pyramidale of CA1 and stratum radiatum of CA2 and by 17-27% throughout most of the rest of CA1, CA2, and CA3. Few, if any, differences were seen between kindled and control animals at 7 or 28 days after the last kindled seizure. Delta binding sites were labeled with 125I-[D-Ala2,D-Leu5]enkephalin in the presence of the morphiceptin analog PL-032. Twenty-four hours after the last kindled seizure, delta binding was decreased only in stratum moleculare of the dentate gyrus. Seven days after the last kindled seizure, delta binding was decreased by 11-17% throughout CA1, CA3, and the dentate gyrus. At 28 days after the last seizure, however, no differences were found between kindled and control animals. Since the decreases in mu and delta opioid binding are transient, they are unlikely to be the molecular basis of the permanent kindling phenomenon. Rather, these changes in opioid binding may represent responses to repeated seizures.


Maternal weight and infections in early childhood: a cohort study.
The aim of this study was to examine this association between maternal weight during pregnancy and the incidence of hospitalisations for infectious diseases during early childhood. A population-based cohort study. A national cohort was created by combining data from the Swedish Medical Birth Register, the National Inpatient Register, the Cause of Death Register, the Total Population Register and the Longitudinal integration database for health insurance and labour market studies. 693 007 children born in Sweden between 1998 and 2006. Number of hospitalisations for infectious diseases during the first 5 years of life, overall and for categories of infectious diseases (lower respiratory, enteric, upper respiratory, genitourinary, perinatal, skin and soft tissue, neurological and eye, digestive tract, bloodstream and other infections). Overweight (body mass index (BMI) 25.0-29.9) and obesity (BMI≥30) during pregnancy were associated with a higher overall incidence of hospitalisations for infectious diseases, adjusted incidence rate ratio (IRR) 1.05 (95% CI 1.03 to 1.06) and adjusted IRR 1.18 (95% CI 1.16 to 1.21). Overweight and obesity during pregnancy were strongly associated with perinatal infections, adjusted IRR 1.34 (95% CI 1.25 to 1.44) and adjusted IRR 1.72 (95% CI 1.57 to 1.88). In contrast, we found no association between maternal weight during pregnancy and infections of skin and soft tissue, the nervous system, the digestive tract or the bloodstream. We observed an association between overweight and obesity during pregnancy, and hospitalisations for infectious diseases during early childhood.


Endoscope-Assisted Extreme-Lateral Interbody Fusion: Preliminary Experience and Technical Note.
The extreme lateral lumbar interbody fusion (XLIF) technique is safe and effective; however, the deep and tight surgical corridor makes visual identification of important landmark structures, as well as sufficient endplate and contralateral preparation, challenging. In the present study, we analyzed the safety and feasibility of endoscope-assisted (EA) XLIF procedures. This was a retrospective single-center study on consecutive patients undergoing XLIF procedures between February 2014 and July 2016. EA-XLIF and conventional XLIF (c-XLIF) procedures were compared in terms of the duration of surgery, estimated blood loss (EBL), perioperative and postoperative complications, and postoperative outcomes. A total of 41 patients (mean age, 66.7 years ± 10.0 years; 22 males [53.7%]) underwent a XLIF procedure, including 6 (14.6%) who underwent EA-XLIF. EA-XLIF did not increase the duration of surgery or EBL. No perioperative or postoperative complications were observed in any of the EA-XLIF procedures. Clinical and radiologic outcomes at 6 weeks postsurgery and at the last follow-up (mean, 8.0 ± 5.8 months postsurgery) were similar for patients in the EA-XLIF and c-XLIF groups. The EA-XLIF technique was considered particularly helpful for checking the lumbar plexus anatomy on the psoas surface, identifying the relationship between the peritoneum and the psoas muscle, positioning the shim into the disc space, removing the disk, and checking the quality of contralateral release and endplate preparation. The EA-XLIF technique is safe and may be considered as an adjunct procedure, offering improved visualization to guide the surgeon in key steps of the XLIF procedure.


Characterization of murine erythroid progenitors with high erythropoietin sensitivity in vitro.
The microplasma clot culture system was employed to study the characteristics of late murine erythroid progenitor cells. Erythroid cluster and colony-forming cells (CFUE) were compared with regard to erythropoietin sensitivity, cell-cycle activity, response to hypertransfusion-induced plethora, average cell size, temporal kinetics of in vitro cluster or colony formation, and relative frequency in murine bone marrow. Results indicate that erythroid cluster-forming cells represent a stage of differentiation which is further along the erythroid pathway than CFUE and are nearly 6-fold more sensitive to the hormone.


The effects of anxiety and strategic planning on visual search behaviour.
The past decade has witnessed increased interest in the visual search behaviour of athletes. Little is known, however, about the relationship between anxiety and eye movements in sport performers or about the extent to which athletes' planned and actual visual search strategies correspond. To address these issues, we conducted two studies. In Study 1, eight expert female gymnasts were presented with three digital slides of a model performing a skill that is known to be anxiety-provoking in this sport--namely, the 'back flip' on the beam. By varying the height of the beam and the presence or absence of safety mats, the slides differed in the amount of anxiety that they elicited vicariously in the viewer. In the study, the gymnasts were asked to imagine themselves in the position of the depicted model and to describe the anxiety that they felt. As they viewed the slides, their eye movements were recorded. As predicted, anxiety was associated with an increase in the number of fixations to peripheral areas. In addition, the more 'threatening' slides elicited significantly more fixations than the less feared images. In Study 2, the plans of 15 equestrian performers (5 expert, 5 intermediate and 5 novice) were elicited as they engaged in a virtual 'walk' around a computerized show-jumping course. Contrary to expectations, the congruence between intended and actual search behaviour was not significantly greater for expert riders than for the less skilled groups. Also, the fact that the top riders allocated more fixations to slides than the less skilled performers challenged the prediction that expertise would be associated with economy of visual search. Finally, as expected, the expert riders were significantly less dependent on the overall 'course plan' than the intermediate and novice equestrian performers when inspecting the fences.


Ultrasound biomicroscopy of globes from young adult pigs.
To determine anterior chamber ocular measurements of adult porcine globes without histologic fixation by use of ultrasound biomicroscopy scanning. 25 porcine globes obtained at an abattoir. Globes were packed on ice for transport. In the laboratory, globes were trimmed, rinsed with antibiotic solution, secured on a single gauze-fold in a latex holder, then were ultrasonogrammed unreformed. Ultrasound biomicroscopy scanning was done, using a 50-MHz transducer, 17-mm cup, and 2% methyl cellulose. Average young adult pig external ocular measurements were: nasal-temporal corneal diameter, 16.61 mm; superior-inferior corneal diameter, 14.00 mm; nasal-temporal globe diameter, 25.48 mm; superior-inferior globe diameter, 24.48 mm; and axial length 21.64 mm. Ultrasound biomicroscopy anterior chamber measurements were: iris sulcus, 30.45 degrees; ciliary sulcus, 18.89 degrees; central corneal thickness, 0.98 mm; corneal thickness at limbus, 1.19 mm; central iris thickness, 0.58 mm; iris tip to ciliary apex, 1.73 mm; iris tip to iris sulcus origin, 3.83 mm; iris tip to ciliary sulcus origin, 2.98 mm; anterior chamber depth from iris tip to cornea, 2.21 mm; central anterior chamber depth, 2.47 mm; ciliary process mid-thickness, 0.65 mm; ciliary process apex to origin of iris sulcus, 2.32 mm; ciliary process apex to origin of ciliary sulcus, 1.34 mm; zonular bundle diameter, 0.10 mm; and interzonular bundle space, 0.11 mm. Anatomic anterior chamber measurements and relations in porcine globes can be used to describe trauma, confirm existence of lesions, and help explain theory. Ultrasound biomicroscopy is a clinical decision aid facilitating noninvasive anatomic or pathologic description without histologic fixation.


Enzymatic Reconstitution and Biosynthetic Investigation of the Lasso Peptide Fusilassin.
Lasso peptides are a class of ribosomally synthesized and post-translationally modified natural product which possess a unique lariat knot conformation. The low entropy "threaded" conformation endows lasso peptides with considerable resistance to heat and proteolytic degradation, which are attractive properties for the development of peptide-based therapeutics. Despite their discovery nearly 30 years ago, the molecular mechanism underlying lasso peptide biosynthesis remains poorly characterized due to the low stability of the purified biosynthetic enzymes. Here, we report the biosynthetic reconstitution of a lasso peptide derived from Thermobifida fusca, termed fusilassin. Beyond robust catalytic activity, the fusilassin enzymes demonstrate extraordinary substrate tolerance during heterologous expression in E. coli and upon purification in cell-free biosynthetic reconstitution reactions. We provide evidence that the fusilassin biosynthetic enzymes are not capable of forming branched-cyclic products but can produce entirely unrelated lasso peptides. Finally, we leveraged our bioinformatic survey of all lasso peptides identified in GenBank to perform coevolutionary analysis of two requisite biosynthetic proteins. This effort correctly identified residues governing an important protein-protein interaction, illustrating how genomic insight can accelerate the characterization of natural product biosynthetic pathways. The fusilassin enzymes described within represent a model system for both designing future lasso peptides of biomedical importance and also for elucidating the molecular mechanisms that govern lasso peptide biosynthesis.


Mechanism-based selection of stabilization strategy for amorphous formulations: Insights into crystallization pathways.
We developed a step-by-step experimental protocol using differential scanning calorimetry (DSC), dynamic vapour sorption (DVS), polarized light microscopy (PLM) and a small-scale dissolution apparatus (μDISS Profiler) to investigate the mechanism (solid-to-solid or solution-mediated) by which crystallization of amorphous drugs occurs upon dissolution. This protocol then guided how to stabilize the amorphous formulation. Indapamide, metolazone, glibenclamide and glipizide were selected as model drugs and HPMC (Pharmacoat 606) and PVP (K30) as stabilizing polymers. Spray-dried amorphous indapamide, metolazone and glibenclamide crystallized via solution-mediated nucleation while glipizide suffered from solid-to-solid crystallization. The addition of 0.001%-0.01% (w/v) HPMC into the dissolution medium successfully prevented the crystallization of supersaturated solutions of indapamide and metolazone whereas it only reduced the crystallization rate for glibenclamide. Amorphous solid dispersion (ASD) formulation of glipizide and PVP K30, at a ratio of 50:50% (w/w) reduced but did not completely eliminate the solid-to-solid crystallization of glipizide even though the overall dissolution rate was enhanced both in the absence and presence of HPMC. Raman spectroscopy indicated the formation of a glipizide polymorph in the dissolution medium with higher solubility than the stable polymorph. As a complementary technique, molecular dynamics (MD) simulations of indapamide and glibenclamide with HPMC was performed. It was revealed that hydrogen bonding patterns of the two drugs with HPMC differed significantly, suggesting that hydrogen bonding may play a role in the greater stabilizing effect on supersaturation of indapamide, compared to glibenclamide.


Ionic conductances of squid giant fiber lobe neurons.
The cell bodies of the neurons in the giant fiber lobe (GFL) of the squid stellate ganglion give rise to axons that fuse and thereby form the third-order giant axon, whose initial portion functions as the postsynaptic element of the squid giant synapse. We have developed a preparation of dissociated, cultured cells from this lobe and have studied the voltage-dependent conductances using patch-clamp techniques. This system offers a unique opportunity for comparing the properties and regional differentiation of ionic channels in somatic and axonal membranes within the same cell. Some of these cells contain a small inward Na current which resembles that found in axon with respect to tetrodotoxin sensitivity, voltage dependence, and inactivation. More prominent is a macroscopic inward current, carried by Ca2+, which is likely to be the result of at least two kinetically distinct types of channels. These Ca channels differ in their closing kinetics, voltage range and time course of activation, and the extent to which their conductance inactivates. The dominant current in these GFL neurons is outward and is carried by K+. It can be accounted for by a single type of voltage-dependent channel. This conductance resembles the K conductance of the axon, except that it partially inactivates during relatively short depolarizations. Ensemble fluctuation analysis of K currents obtained from excised outside-out patches is consistent with a single type of K channel and yields estimates for the single channel conductance of approximately 13 pS, independently of membrane potential. A preliminary analysis of single channel data supports the conclusion that there is a single type of voltage-dependent, inactivating K channel in the GFL neurons.


The Impact of Epigenetics on Mesenchymal Stem Cell Biology.
Changes in epigenetic marks are known to be important regulatory factors in stem cell fate determination and differentiation. In the past years, the investigation of the epigenetic regulation of stem cell biology has largely focused on embryonic stem cells (ESCs). Contrarily, less is known about the epigenetic control of gene expression during differentiation of adult stem cells (AdSCs). Among AdSCs, mesenchymal stem cells (MSCs) are the most investigated stem cell population because of their enormous potential for therapeutic applications in regenerative medicine and tissue engineering. In this review, we analyze the main studies addressing the epigenetic changes in MSC landscape during in vitro cultivation and replicative senescence, as well as follow osteocyte, chondrocyte, and adipocyte differentiation. In these studies, histone acetylation, DNA methylation, and miRNA expression are among the most investigated phenomena. We describe also epigenetic changes that are associated with in vitro MSC trans-differentiation. Although at the at initial stage, the epigenetics of MSCs promise to have profound implications for stem cell basic and applied research. J. Cell. Physiol. 231: 2393-2401, 2016. © 2016 Wiley Periodicals, Inc.


Water absorption of poly(methyl methacrylate) measured by vertical interference microscopy.
PMMA (poly(methyl methacrylate)) is widely used to prepare orthopedic cements. They are in direct contact with cells and body fluids. PMMA, despite its hydrophobic nature, can absorb ~2% w/w water. We have evaluated by vertical interference microscopy if water absorption can produce a significant swelling in different types of PMMA blocks: pure, with a plasticizer, with a cross-linker, and in two types of commercial bone cements. Graphite rods which do not swell in water were used as internal standard. Hardness, indentation modulus, plastic, and elastic works were determined by nanoindentation under a 25mN fixed force. Vertical interference microscopy was used to image the polymer in the dry state and hydrated states (after 24 h in distilled water). On the surface of the polished polymers (before and after hydration), we measured roughness by the fractal dimension, the swelling in the vertical and the lateral directions. For each polymer block, four images were obtained and values were averaged. Comparison and standardization of the images in the dry and hydrated states were done with Matlab software. The average value measured on the graphite rod between the two images (dried and hydrated) was used for standardization of the images which were visualized in 3D. After grinding, a small retraction was noticeable between the surface of the rod and the polymers. A retraction ring was also visible around the graphite rod. After hydration, only the pure PMMA and bone cements had a significant swelling in the vertical direction. The presence of polymer beads in the cements limited the swelling in the lateral direction. Swelling parameters correlated with the nanoindentation data. PMMA can swell by absorbing a small amount of water and this induces a swelling that varies with the polymer composition and particle inclusions.


Second-Trimester Ultrasound-Measured Umbilical Cord Insertion-to-Placental Edge Distance: Determining an Outcome-Based Threshold for Identifying Marginal Cord Insertions.
We sought to study potential diagnostic criteria for marginal placental cord insertions as determined by associations with obstetric outcomes. This single-center retrospective cohort investigation included singleton gestations delivering from January 1, 2012, to December 31, 2016, and having a standard or detailed fetal ultrasound examination from 18 weeks to 21 weeks 6 days. Cord insertion-to-placental edge distances were used to create a referent comparison group and 3 patient groups: greater than 3.0 cm (comparison); greater than 2.0 to 3.0 cm (group 1); greater than 1.0 to 2.0 cm (group 2); and 1.0 cm or less (group 3). The primary outcome consisted of any one of spontaneous delivery before 37 weeks, fetal growth restriction, oligohydramnios, placental abruption, or intrauterine fetal demise. The numbers of participants and mean distances ± SDs for the comparison group and groups 1 to 3, respectively, were 628 and 4.68 ± 1.17 cm, 106 and 2.52 ± 0.26 cm, 131 and 1.60 ± 0.29 cm, and 77 and 0.36 ± 0.37 cm, respectively. The primary outcome was significantly associated only with cord insertion-to-placental edge distances of 1.0 cm or less (adjusted odds ratio, 3.05; 95% confidence interval, 1.73-5.38). Marginal cord insertions may be diagnosed when the cord insertion-to-placental edge distance is 1.0 cm or less.


Changes in soft tissue dimensions following three different techniques of stage-two surgery: a case series report.
The aim of this case series report is to compare the results of the increase in keratinized mucosa using three different techniques of stage-two surgery. Thirty-two patients with one to eight dental implants who received prosthetic rehabilitation of the maxilla were included. Patients were divided into three groups based on preoperative anatomical considerations. Stage-two surgery was performed using either the apically repositioned flap (ARF; n = 14), the roll flap (RF; n = 10), or an apically repositioned flap combined with a connective tissue graft (ARFCT; n = 8). The height of the keratinized mucosa and relative tissue thickness were measured preoperatively and postoperatively at 2 weeks and 3, 6, and 12 months after surgery. The mean gains of keratinized mucosa and tissue thickness were calculated from these measurements. After 1 year, the mean gains in tissue thickness and keratinized tissue were 1.37 and 4.63 mm in the ARF group, 2.41 and 1.35 mm in the RF group, and 3.10 and 4.10 mm in the ARFCT group, respectively. There was no significant statistical difference between the 12-month and postoperative measurements (P > .05). In patients with deficient tissue thickness, a roll flap or an apically repositioned flap should be performed, while a lack of keratinized mucosa indicates the use of an apically repositioned flap with or without a connective tissue graft. When an increase in both keratinized mucosa and tissue thickness is necessary, an apically repositioned flap combined with a free connective tissue graft can be recommended. After a 12-month healing period, the obtained results showed excellent stability.


Detecting scaling in the period dynamics of multimodal signals: application to Parkinsonian tremor.
Patients with Parkinson's disease exhibit tremor, involuntary movement of the limbs. The frequency spectrum of tremor typically has broad peaks at "harmonic" frequencies, much like that seen in other physical processes. In general, this type of harmonic structure in the frequency domain may be due to two possible mechanisms: a nonlinear oscillation or a superposition of (multiple) independent modes of oscillation. A broad peak spectrum generally indicates that a signal is semiperiodic with a fluctuating period. These fluctuations may posses intrinsic order that can be quantified using scaling analysis. We propose a method to extract the correlation (scaling) properties in the period dynamics of multimodal oscillations, in order to distinguish between a nonlinear oscillation and a superposition of individual modes of oscillation. The method is based on our finding that the information content of the temporal correlations in a fluctuating period of a single oscillator is contained in a finite frequency band in the power spectrum, allowing for decomposition of modes by bandpass filtering. Our simulations for a nonlinear oscillation show that harmonic modes possess the same scaling properties. In contrast, when the method is applied to tremor records from patients with Parkinson's disease, the first two modes of oscillations yield different scaling patterns, suggesting that these modes may not be simple harmonics, as might be initially assumed.


Meningitis following gunshot wound of the neck.
It is generally assumed that a missile fired from a gun is subjected to sufficient heat to render it sterilized. For this reason, retained bullets are not usually considered a source of infection. The infectious complications associated with gunshot wounds are typically attributed to perforation of a hollow viscus with leakage of gastrointestinal contents causing peritonitis or intra-abdominal abscess. There are several reports of bacterial meningitis involving the spinal cord in gunshot wounds that perforate the intestine prior to involving the thoracic or lumbar vertebral column; however, there are no published reports of cerebral meningitis resulting from a retained projectile in the spinal canal in which there was no injury to the gastrointestinal tract. This manuscript describes a woman who died as a result of unsuspected acute bacterial meningitis which developed secondary to a gunshot wound of the neck. The projectile fractured the first thoracic vertebra, lacerated the dura and contused the spinal cord at the C7-T1 junction. Meningitis developed at the C7-T1 level and ascended along the cervical spinal cord to the brain. The infection caused acute neurologic deterioration and death four days following the initial injury.


Synthesis of bioactive and stabilized cyclic peptides by macrocyclization using C(sp3)-H activation.
Cyclic peptides have attracted increasing attention in recent years due to their ability to inhibit protein-protein interactions. Current strategies to prepare cyclic peptides often rely on functional amino acid side chains or the incorporation of unnatural amino acids, thus limiting their structural diversity. Here, we describe the development of a highly versatile peptide macrocyclization strategy through a palladium-catalyzed C(sp3)-H activation and the synthesis of cyclic peptides featuring unique hydrocarbon linkages between the β-carbon of amino acids and the aromatic side chains of Phe and Trp. We demonstrate that such peptides exhibit improved biological properties compared to their acyclic counterparts. Finally, we applied this method in the synthesis of the natural product celogentin C.


Regression of fibrosis and hypertrophy in failing myocardium following mechanical circulatory support.
The cellular and structural changes that occur during long-term ventricular unloading leading to cardiac recovery are poorly understood. However, we have previously demonstrated that left ventricular assist device (LVAD) support leads to a significant decrease in intracardiac tumor necrosis factor-alpha (TNF-alpha), a protein capable of producing hypertrophy and fibrosis. To further define the beneficial effects of long-term ventricular unloading on cardiac function, we determined the effect of mechanical circulatory support on fibrosis and hypertrophy in paired myocardial samples of 18 patients with end-stage cardiomyopathy obtained at the time of LVAD implantation and removal. We determined total collagen as well as collagen I and III by a semiquantitative analysis of positive immune-stained areas in pre- and post-LVAD myocardial samples. We found that total collagen content was reduced by 72% (p < 0.001), whereas collagen I content decreased by 66% (p < 0.001) and collagen III content was reduced by 62% (p < 0.001). Next, we determined myocyte size by direct analysis of cellular dimensions utilizing a computerized edge detection system in pre- and post-LVAD myocardial samples. We found that myocyte size decreased in all patients studied for an average reduction of 26% (33.1 +/- 1.32 to 24.4 +/- 1.64 microm, p < 0.001). These data demonstrate that long-term mechanical circulatory support significantly reduces collagen content and decreases myocyte size. We suggest that the reduction of fibrosis and hypertrophy observed may in part contribute to the recovery of cardiac function associated with long-term mechanical circulatory support.


Patients' and staff's perceptions of the psychiatric ward environment.
This study examined the extent to which patients' and staff's perceptions of the psychiatric ward atmosphere and the working conditions of staff influence patient and staff satisfaction. A total of 640 staff members on 42 wards completed the Ward Atmosphere Scale (WAS) and the Working Environment Scale-10 (WES-10). A total of 424 patients on the same wards completed the WAS. Both patients and staff members answered three questions about their general satisfaction with the ward. The ward means were used as the unit of analysis. The staff members had significantly higher scores than the patients on nine of the 11 WAS subscales. Patients' and staff's WAS scores were moderately correlated. No significant correlation was found between patients' and staff's scores on the three satisfaction items. Patient satisfaction was strongly correlated with patients' WAS scores and was moderately correlated with staff's WAS scores but was not significantly correlated with staff's WES-10 scores. Staff satisfaction was moderately correlated with staff's WAS and WES-10 scores but was not correlated with patients' WAS scores. Different aspects of the treatment climate seem to be important for patient and staff satisfaction. The ward atmosphere seems to be more important for patient satisfaction than for staff satisfaction. The working environment is strongly related to staff satisfaction but seems to be unrelated to patient satisfaction.


A Stable Genetic Transformation System and Implications of the Type IV Restriction System in the Nitrogen-Fixing Plant Endosymbiont Frankia alni ACN14a.
Genus Frankia is comprised primarily of nitrogen-fixing actinobacteria that form root nodule symbioses with a group of hosts known as the actinorhizal plants. These plants are evolutionarily closely related to the legumes that are nodulated by the rhizobia. Both host groups utilize homologs of nodulation genes for root-nodule symbiosis, derived from common plant ancestors. The corresponding endosymbionts, Frankia and the rhizobia, however, are distantly related groups of bacteria, leading to questions about their symbiotic mechanisms and evolutionary history. To date, a stable system of electrotransformation has been lacking in Frankia despite numerous attempts by research groups worldwide. We have identified type IV methyl-directed restriction systems, highly-expressed in a range of actinobacteria, as a likely barrier to Frankia transformation. Here we report the successful electrotransformation of the model strain F. alni ACN14a with an unmethylated, broad host-range replicating plasmid, expressing chloramphenicol-resistance for selection and GFP as a marker of gene expression. This system circumvented the type IV restriction barrier and allowed the stable maintenance of the plasmid. During nitrogen limitation, Frankia differentiates into two cell types: the vegetative hyphae and nitrogen-fixing vesicles. When the expression of egfp under the control of the nif gene cluster promoter was localized using fluorescence imaging, the expression of nitrogen fixation in nitrogen-limited culture was localized in Frankia vesicles but not in hyphae. The ability to separate gene expression patterns between Frankia hyphae and vesicles will enable deeper comparisons of molecular signaling and metabolic exchange between Frankia-actinorhizal and rhizobia-legume symbioses to be made, and may broaden potential applications in agriculture. Further downstream applications are possible, including gene knock-outs and complementation, to open up a range of experiments in Frankia and its symbioses. Additionally, in the transcriptome of F. alni ACN14a, type IV restriction enzymes were highly expressed in nitrogen-replete culture but their expression strongly decreased during symbiosis. The down-regulation of type IV restriction enzymes in symbiosis suggests that horizontal gene transfer may occur more frequently inside the nodule, with possible new implications for the evolution of Frankia.


The Prader-Willi/Angelman imprinted domain and its control center.
The present review focuses on the recent advances towards understanding the mode of operation of the imprinting center (IC) within the Prader-Willi/Angelman syndromes (PWS/AS) domain. Special emphasis is put on the elucidation of the functional interaction between the two parts of the center, AS-IC and PWS-IC. The recent studies, on which the review is based, reveal cis-acting elements and trans-acting proteins that constitute the two parts of the IC and presumably provide the molecular mechanism for this interaction. AS-IC acquires the primary imprint during gametogenesis by establishing the maternal epigenotype. The unmethylated maternal allele of the AS-IC binds, very likely, a trans-acting factor that confers methylation on the PWS-IC maternal allele after fertilization. It is assumed that the PWS-IC paternal epigenotype, once established, spreads across the entire PWS/AS domain in the soma.


Prevalence and characteristics of multinucleoside-resistant human immunodeficiency virus type 1 among European patients receiving combinations of nucleoside analogues.
The prevalence and the genotypic and phenotypic characteristics of multinucleoside-resistant (MNR) human immunodeficiency virus type 1 (HIV-1) variants in Europe were investigated in a multicenter study that involved centers in nine European countries. Study samples (n = 363) collected between 1991 and 1997 from patients exposed to two or more nucleoside analogue reverse transcriptase inhibitors (NRTIs) and 274 control samples from patients exposed to no or one NRTI were screened for two marker mutations of multinucleoside resistance (the Q151M mutation and a mutation with a 2-amino-acid insertion at codon 69, T69S-XX). Q151M was identified in six of the study samples (1. 6%), and T69S-XX was identified in two of the study samples (0.5%; both of them T69S-SS), but both patterns were absent among control samples. Non-NRTI (NNRTI)-related changes were observed in viral strains from two patients, which displayed the Q151M resistance pattern, although the patients were NNRTI naive. The patients whose isolates displayed multinucleoside resistance had received treatment with zidovudine and either didanosine, zalcitabine, or stavudine. Both resistance patterns conferred broad cross-resistance to NRTIs in vitro and a poor response to treatment in vivo. MNR HIV-1 is found only among multinucleoside-experienced patients. Its prevalence is low in Europe, but it should be closely monitored since it seriously limits treatment options.


alpha 1-antitrypsin TAQ I polymorphism and alpha 1-antichymotrypsin mutations in patients with obstructive pulmonary disease.
Obstructive pulmonary disease is a multifactorial condition deriving from the interaction of environmental and genetic factors. From biochemical knowledge of the basis of the disease, alpha 1-antitrypsin and alpha 1-antichymotrypsin are considered two likely candidate genes. We therefore designed an association study comprising 232 unrelated Italian individuals divided as follows: 89 individuals with obstructive lung disease (66 with COPD and 23 with disseminated bronchiectasis) and 143 controls (45 patients with non-obstructive lung disease and 98 healthy individuals). We screened for Taq I (G1237A) polymorphism of the alpha 1-antitrypsin gene as well as the rare variants Bonn-1 (Pro229Ala), Bochum-1 (Leu55Pro), Isehara-1 (Met389Val) and Isehara-2 (1258delAA), and the common signal peptide polymorphism Thr-15Ala of the alpha 1-antichymotrypsin gene. The frequencies of Taq I G1237A alleles were 11.7 and 10.8% in obstructed patients and controls, respectively (P = 0.43), while those of signal peptide Thr-15Ala alleles were 51.6 and 50.3% in obstructed patients and controls, respectively (P = 0.42). We conclude that alpha 1-antitrypsin Taq I polymorphism and alpha 1-antichymotrypsin Thr-15Ala mutation are not major genetic risk factors for the development of obstructive lung disease in Italian patients. The alpha 1-antichymotrypsin rare variants were not detected: our results do not exclude the possibility that other alpha 1-antichymotrypsin gene mutations might be present in Italian obstructed patients but, if so, these genetic defects must be rare.


Static Magnetic Field Reduces Intracellular ROS Levels and Protects Cells Against Peroxide-Induced Damage: Suggested Roles for Catalase.
A feature in neurodegenerative disorders is the loss of neurons, caused by several factors including oxidative stress induced by reactive oxygen species (ROS). In this work, static magnetic field (SMF) was applied in vitro to evaluate its effect on the viability, proliferation, and migration of human neuroblastoma SH-SY5Y cells, and on the toxicity induced by hydrogen peroxide (H2O2), tert-butyl hydroperoxide (tBHP), H2O2/sodium azide (NaN3) and photosensitized oxidations by photodynamic therapy (PDT) photosensitizers. The SMF increased almost twofold the cell expression of the proliferation biomarker Ki-67 compared to control cells after 7 days of exposure. Exposure to SMF accelerated the wound healing of scratched cell monolayers and significantly reduced the H2O2-induced and the tBHP-induced cell deaths. Interestingly, SMF was able to revert the effects of NaN3 (a catalase inhibitor), suggesting an increased activity of catalase under the influence of the magnetic field. In agreement with this hypothesis, SMF significantly reduced the oxidation of DCF-H2, indicating a lower level of intracellular ROS. When the redox imbalance was triggered through photosensitized oxidation, no protection was observed. This observation aligns with the proposed role of catalase in cellular proctetion under SMF. Exposition to SMF should be further validated in vitro and in vivo as a potential therapeutic approach for neurodegenerative disorders.


Oncological outcomes and failure patterns of laser cordectomy in recurrent glottic cancer.
Laser cordectomy is a widely accepted treatment modality for selected cases of early glottic cancers, but its role as a salvage treatment remains unclear. We aimed to investigate the oncological outcomes and failure patterns of salvage cordectomy. This was a retrospective cohort study of patients who underwent cordectomy for early glottic cancer between 2013 and 2022 at a tertiary referral center. The main outcome measures were overall survival, larynx-preservation rate, tracheostomy dependency rate, and disease-free survival. A total of 142 patients (mean age: 63 years, interquartile range [IQR]: 45-72, 123[86.9 %] males) were analyzed. There were 38 (26.8 %) recurrences after a mean of 22 months (IQR: 17-26). Among them, 25 (17.6 %) underwent salvage cordectomy, while 13 (9.1 %) received other salvage treatments (11[7.7 %] (chemo)radiotherapy and 2[1.4 %] total laryngectomy). In comparison to the other salvage treatments, salvage cordectomy demonstrated lower tracheostomy rates (0 vs. 31 %, p = 0.05), comparable 5-year disease-free survival (62 % vs. 54 %, p = 0.4), higher 5-year larynx preservation rate (92 % vs. 54 %, p = 0.02), and improved 5-year overall survival rate (84 % vs. 62 %, p = 0.01). Factors associated with salvage cordectomy failure were age >60 years (odds ratio [OR]: 1.3, 95 % confidence interval [CI]: 1.15-1.53), smoking continuation (OR: 3.73, 95 % CI: 3.5-4.4), heavy smoking (OR: 1.24, 95 % CI:1.07-2.15), and pT1b + (OR: 2.26, 95 % CI: 2.1-2.9). Salvage cordectomy offers favorable larynx preservation rates and oncological outcomes for recurrent disease amenable to conservative surgery. Smoking, advanced age, and advanced tumor stages are associated with salvage cordectomy failure.


Increased risk of severe maternal morbidity (near-miss) among immigrant women in Sweden: a population register-based study.
To determine if immigrant women from low-, middle- and high-income countries have an increased risk of severe maternal morbidity (near-miss) when they deliver in Sweden. Population register-based study. Nationwide study including all singleton deliveries (≥28 weeks of gestation) between 1998 and 2007. Women with a near-miss event; all women with a singleton delivery ≥28 weeks of gestation during the same period acted as reference group. Near-miss was defined by a combined clinical and management approach with use of International Classification of Diseases, 10th revision codes for severe maternal morbidity. A woman's country of origin was designated as low-, middle- or high-income according to the World Bank Classification of 2009. Unconditional logistic regression models were used in the analysis. Maternal near-miss frequencies per 1000 deliveries and odds ratios with 95% confidence intervals. There were 914 474 deliveries during the study period and 2655 near-misses (2.9 per 1000 deliveries). In comparison to Swedish-born women, those from low-income countries had an increased risk of near-miss (odds ratio 2.3, 95% confidence interval 1.9-2.8) that was significant in all morbidity groups except for cardiovascular diseases and sepsis. Women from middle- and high-income countries showed no increased risk of near-miss. Women from low-income countries have an increased risk of maternal near-miss morbidity compared with women born in Sweden. Although the rate is low it should alert healthcare providers.


Specific differences in temporal binding aspects of the attentional blink in Chromosome 22q11.2 Deletion Syndrome.
Chromosome 22q11.2 Deletion Syndrome (22q11DS) is a genetic syndrome characterized by a variety of cognitive impairments, including difficulty with attention. 22q11DS is the strongest known genetic risk factor for developing schizophrenia, a disorder characterized by impairments in visual attention and temporal binding processes. Here we examine a specific temporal visual attention phenomenon (the attentional blink; AB) within two rapid serial visual presentation tasks, and compare those with 22q11DS to groups of typically developing individuals matched on chronological (CA) and mental age (MA). Performance of individuals with 22q11DS was sensitive to differing task demands. On a Category Task, individuals with 22q11DS performed similarly to control groups on all measures of the AB, with the exception of lower detection accuracy of the first of two targets. In contrast, on a feature-based Color Task which required temporal binding of stimulus features, individuals with 22q11DS differed from CA and MA matched control groups on all AB performance measures, exhibiting lower target accuracy, more temporal binding errors, and a deeper, more protracted AB. Temporal binding in the visual domain is thought to be dependent on a serial attention mechanism that facilitates simultaneous firing of neurons in multiple areas of the visual cortex, activating short-term working memory for storage of bound features. Given the discrepancy between these two tasks, results suggest that temporal binding processes may be significantly affected in individuals with 22q11DS, a finding that importantly, has been previously demonstrated among individuals with schizophrenia.