Casein and alpha-lactalbumin detection in breast cancer cells by immunocytochemistry.
Casein and alpha-lactalbumin are characteristic proteins produced by normal mammary cells under hormonal stimulation. Specific antisera were used to immunochemically stain for these two proteins in the unstimulated R3230AC transplantable rat mammary tumor as well as in normal lactating rat mammary tissue. Specific staining was observed in the lactating rat alveolar epithelial cells and in the cells lining the intraalveolar ducts with both antisera. In the R3230AC tumor some cells were shown to contain casein or alpha-lactalbumin; the majority of the tumor cells were unstained. These findings indicate that normal mammary cells as well as a small population of cells within the R3230AC tumor are actively synthesizing casein or alpha-lactalbumin. Furthermore, the results suggest that immunocytochemistry may be used to determine the functional heterogeneity of mammary tumors directly.


Predictors of suicide attempts in 3.322 patients with affective disorders and schizophrenia spectrum disorders.
This study explores risk factors for suicide attempts using the electronic health records of 3322 patients with either schizophrenia spectrum disorders or affective disorders who underwent a comprehensive psychiatric evaluation at the Emergency Department at the Long Island Jewish Medical Center or the Hillside Evaluation Center at The Zucker Hillside Hospital from August 3rd 2011 to July 5th 2012. Multivariate regression analyses showed, after adjusting for sex, that previous suicidal attempts and financial or relationship losses were significantly associated with a current suicidal attempt. Additionally, higher odds of having a suicidal attempt were also found in those subjects with a diagnosis of an affective disorder, compared to a schizophrenia spectrum diagnosis, and those patients in the children/adolescent group compared to those in the adult/elderly group. Our study results confirm and expand results from prior studies. Therefore, physicians should be alert for the presence of any or all of these factors upon evaluation of psychiatric patients, and if present, either psychiatric hospitalization or a close psychiatric follow up in collaboration with family and a therapist would be key in reducing the risk of potential suicidal behavior.


New insights into acne pathogenesis: propionibacterium acnes activates the inflammasome.
The precise contribution of the commensal bacterium Propionibacterium acnes (P. acnes) in the inflammatory response associated with acne vulgaris remains controversial. In this issue Qin et al. show that P. acnes induces robust IL-1β secretion in monocytic cells by triggering the activation of the NLRP3 inflammasome. In vivo, the encounter of P. acnes and macrophages in the peri-follicular dermis could locally result in the release of substantial amounts of IL-1β and therefore exacerbate inflammation. Such findings suggest that molecules targeting IL-1β and/or the NLRP3 inflammasome may constitute new treatment possibilities for acne vulgaris.


Association of Mental Health Status With Perception of Nasal Function.
Mental health issues are thought to be overrepresented among patients undergoing rhinoplasty and may be associated with patient presentation prior to surgery. To assess the association of poor mental health with perception of nasal function. A cross-sectional study of patients presenting for airway assessment was performed from December 1, 2011, to October 31, 2015, at 2 tertiary rhinoplasty centers in Sydney, Australia. Mental health was independently defined preoperatively by the Mental Component Summary of the 36-item Short Form Health Survey version 2 (a score of <40 indicated poor mental well-being), the Rosenberg Self-Esteem Scale (a score of <15 indicated low self-esteem), and the Dysmorphic Concerns Questionnaire (a score of >11 indicated above-average dysmorphic concerns). Nasal function was assessed with patient-reported outcome measures, including the Nasal Obstruction Symptom Evaluation Scale, the 22-item Sinonasal Outcome Test, a visual analog scale to rate ease of breathing on the left and right sides, and Likert scales to assess overall function and nasal obstruction. Nasal airflow was assessed by nasal peak inspiratory flow, nasal airway resistance, and minimum cross-sectional area. Among 495 patients in the study (302 women and 193 men; mean [SD] age, 36.5 [13.6] years), compared with patients with good mental health, those with poor mental health had poorer scores in all patient-reported outcome measures, including the visual analog scale for the left side (mean [SD], 51 [25] vs 42 [25]; P = .001), visual analog scale for the right side (mean [SD], 54 [24] vs 45 [26]; P < .001), Nasal Obstruction Symptom Evaluation Scale (mean [SD], 2.64 [0.95] vs 1.96 [1.04]; P < .001), 22-item Sinonasal Outcome Test (mean [SD], 2.14 [0.84] vs 1.33 [0.83]; P < .001), nasal obstruction (58 of 145 [40.2%] vs 83 of 350 [23.7%] with severe or worse obstruction; P < .001), and nasal function (72 of 145 [49.7%] vs 111 of 350 [31.8%] with poor or worse function; P < .001). Subclinical differences in nasal peak inspiratory flow could be demonstrated, but all other nasal airflow measures were similar. Low self-esteem produced a similar pattern, but dysmorphia did not. Poor mental health status is associated with a poorer self-perception of nasal function compared with those who are mentally healthy with clinically similar nasal airflow. Clinicians should be aware that patients with poor mental health reporting obstructed airflow may in part be representing an extension of their negative emotions rather than true obstruction and may require further assessment prior to surgery. NA.


The significance of immune microenvironment in patients with endometriosis.
Endometriosis represents an estrogen-dependent disease of the female reproductive system and intra- and extraperitoneal regions, with chronic feature. Currently, immune cells, such as macrophages and lymphocytes, are considered to play a pivotal role in angiogenesis and invasion of endometriotic cells through matrix remodeling. Additionally, various studies have revealed the role of E-cadherin, β-catenin, along with steroid hormone receptors in endometriosis development. In this context, our study aimed to analyze the relationship between the cellular immune profile and E-cadherin, β-catenin, estrogen receptor alpha (ERα), and progesterone receptor (PR) immunoexpression in endometriosis tissues, along with an analysis of the possible association between serological parameters and immunohistochemical (IHC) markers. The study included 53 patients diagnosed with ovarian or cutaneous abdominal wall endometriosis, which have been investigated by routine histology, immunohistochemistry, and serum analysis. The IHC exam showed an increased density of cluster of differentiation (CD)4+ T-cells, CD8+ T-cells, and CD68+ macrophages, along with variable increased expressions of E-cadherin, β-catenin, ERα, and PR. Statistical analysis revealed an intense positive correlation between CD68 and PR expression (p<0.05), without any other statistically significant correlations between IHC markers or between IHC and serological markers. Our study supports that endometriosis is an immune-dependent disease characterized by an abnormal morphological profile of T-cells and macrophages in endometriotic implants. Our study provides additional data useful in the understanding the immune milieu of endometriosis in the context of its complex pathogenic molecular mechanism. Further research is needed to develop new immunological therapeutic approaches, like immune checkpoint inhibitors administration or T-cell-targeted immunotherapy in these patients.


Dietary phytase increases the true absorption and endogenous fecal excretion of zinc in growing pigs given a corn-soybean meal based diet.
We investigated the effect of dietary phytase on the true absorption and endogenous fecal excretion of zinc (Zn) using (67)Zn in growing pigs given a corn-soybean meal based diet. Ten crossbred barrows were fed the control diet containing 90-mg/kg Zn, 2.3-g/kg phytate-phosphorus and 3.7-g/kg non-phytate-phosphorus or the phytase diet containing similar amounts of Zn and phytate-phosphorus, and 1.4-g/kg non-phytate-phosphorus with 750-PU/kg phytase for 12 h/day. On day 6, the pigs were given 200 g of the corresponding diet labeled by (67)Zn for 2 h. We measured feed intake, fecal Zn concentration and (67)Zn abundance for the determination of apparent absorption, true absorption and endogenous fecal excretion of Zn. Although the apparent absorption of Zn did not significantly differ between the dietary groups, the phytase group had significantly more (P < 0.05) true absorption of Zn than the control group. The endogenous fecal excretion of Zn tended to be more (P = 0.07) in the phytase group than in the control group. These results suggest that dietary phytase improves Zn bioavailability through increasing the true absorption of Zn in growing pigs, which results in stimulating the endogenous fecal excretion of Zn when dietary Zn satisfies its requirement.


Histopathologic evaluation of pulpal tissue response to various adhesive cleanup techniques.
The aim of this prospective in-vivo study was to investigate the possible effects of temperature changes from various adhesive cleanup procedures on pulpal tissue. The materials, consisting of 40 sound maxillary and mandibular premolars to be extracted during orthodontic treatment, were randomly assigned to 4 groups, with 1 group as the control. The teeth in the 3 study groups were etched; brackets were bonded and then debonded. The remaining adhesive was removed with a tungsten carbide bur by using a high-speed hand piece. The teeth in the control group were not etched and bonded. In group 1, the residual adhesive was removed with water cooling, and the teeth were extracted 24 hours later. In group 2, the residual adhesive was removed without water cooling, and the teeth were extracted 24 hours later. In group 3, the residual adhesive was removed without water cooling, and the teeth were extracted 20 days later. The teeth were prepared for histologic examination, and the number of vessels, vessel areas and perimeters, extravasation of red blood cells, vascular congestion, and inflammatory cell infiltration were evaluated to determine pulpal tissue changes. According to the findings from histologic and immunohistochemical evaluations, the coronal pulps of the teeth in groups 1 and 3 were almost similar to the control teeth, but some distinct pathologic changes were observed in group 2. Adhesive removal without water cooling caused some vascular and pulpal tissue alterations, but these were tolerated by the pulpal tissues, so the changes were reversible.


Sustainable irrigation based on co-regulation of soil water supply and atmospheric evaporative demand.
Irrigation is an important adaptation to reduce crop yield loss due to water stress from both soil water deficit (low soil moisture) and atmospheric aridity (high vapor pressure deficit, VPD). Traditionally, irrigation has primarily focused on soil water deficit. Observational evidence demonstrates that stomatal conductance is co-regulated by soil moisture and VPD from water supply and demand aspects. Here we use a validated hydraulically-driven ecosystem model to reproduce the co-regulation pattern. Specifically, we propose a plant-centric irrigation scheme considering water supply-demand dynamics (SDD), and compare it with soil-moisture-based irrigation scheme (management allowable depletion, MAD) for continuous maize cropping systems in Nebraska, United States. We find that, under current climate conditions, the plant-centric SDD irrigation scheme combining soil moisture and VPD, could significantly reduce irrigation water use (-24.0%) while maintaining crop yields, and increase economic profits (+11.2%) and irrigation water productivity (+25.2%) compared with MAD, thus SDD could significantly improve water sustainability.


Multiple Nodulation Genes Are Up-Regulated During Establishment of Reniform Nematode Feeding Sites in Soybean.
The semi-endoparastic reniform nematode (Rotylenchulus reniformis) infects over 300 plant species. Females penetrate host roots and induce formation of complex, multinucleate feeding sites called syncytia. While anatomical changes associated with reniform nematode infection are well documented, little is known about their molecular basis. We grew soybean (Glycine max) in a split-root growth system, inoculated half of each root system with R. reniformis, and quantified gene expression in infected and control root tissue at four dates after inoculation. Over 6,000 genes were differentially expressed between inoculated and control roots on at least one date (false discovery rate [FDR] = 0.01, |log2FC| ≥ 1), and 507 gene sets were significantly enriched or depleted in inoculated roots (FDR = 0.05). Numerous genes up-regulated during syncytium formation had previously been associated with rhizobia nodulation. These included the nodule-initiating transcription factors CYCLOPS, NSP1, NSP2, and NIN, as well as multiple nodulins associated with the plant-derived peribacteroid membrane. Nodulation-related NIP aquaporins and SWEET sugar transporters were induced, as were plant CLAVATA3/ESR-related (CLE) signaling proteins and cell cycle regulators such as CCS52A and E2F. Nodulins and nodule-associated genes may have ancestral functions in normal root development and mycorrhization that have been co-opted by both parasitic nematodes and rhizobial bacteria to promote feeding site and nodule formation.


The coordination of contractions of circular and longitudinal muscle layers of rat uterus in late pregnancy and during delivery.
Although it is well known that, in visceral smooth muscle, gap junctions between muscle fibers of the same muscle layer are responsible for synchronized strong contractions, much less attention has been paid to the interaction of muscle layers. An L-shaped preparation of the myometrium composed of circular and longitudinal muscle layers was adapted to study the relationship between the activities of these two muscle layers of pregnant rats. After careful separation, each of the two arms of the preparation would consist of only one muscle layer, while both layers remained connected at the angle of the L. Thus, contractions of both muscle layers could be monitored simultaneously and their relationship could be investigated. It was found that coordination of the contractions of longitudinal and circular muscles appeared in the morning of gestation day 21 and continued toward delivery. The coordination was expressed as a special sequence of the contractions of the two muscle layers which occurred repetitively for an hour or longer. In most of the cases, pacemaker was located in the longitudinal layer. Dissociation of one of the arms would make both arms contract independently. A few specimens from animals in their early third trimester also showed interaction of the muscle layers. Nevertheless, the correlation was not as strong as that in specimens prepared from animals close to term.


Parasites and a host's sense of smell: reduced chemosensory performance of fathead minnows (Pimephales promelas) infected with a monogenean parasite.
Parasites residing within the central nervous system of their hosts have the potential to reduce various components of host performance, but such effects are rarely evaluated. We assessed the olfactory acuity of fathead minnows (Pimephales promelas) infected experimentally with the monogenean Dactylogyrus olfactorius, the adults of which live within the host's olfactory chambers. Olfactory acuity was compared between infected and uninfected hosts by assessing electro-olfactography (EOG) neural responses to chemical stimuli that indicate the presence of food (L-alanine) or the presence of conspecifics (taurocholic acid). We also compared differences in gross morphology of the olfactory epithelium in infected and uninfected minnows. Differences in EOG responses between infected and uninfected minnows to both cue types were non-significant at 30 days post-exposure. By days 60 and 90, coincident with a two times increase in parasite intensity in the olfactory chambers, the EOG responses of infected minnows were 70-90% lower than controls. When infected fish were treated with a parasiticide (Prazipro), olfactory acuity returned to control levels by day 7 post-treatment. The observed reduction in olfactory acuity is best explained by the reduced density of cilia covering the olfactory chambers of infected fish, or by the concomitant increase in the density of mucous cells that cover the olfactory chambers. These morphological changes are likely due to the direct effects of attachment and feeding by individual worms or by indirect effects associated with host responses. Our results show that infection of a commonly occurring monogenean in fathead minnows reduces olfactory acuity. Parasite-induced interference with olfactory performance may reduce a fish's ability to detect, or respond to, chemical cues originating from food, predators, competitors or mates.


Fracture toughness of water-aged resin composite restorative materials.
The purpose of this study was to assess the effects of aging experimental dimethacrylate resin composites in water at 37 degrees C for periods up to 6 wk by measuring the variations in fracture toughness (K(c)), elastic modulus (E), fracture energy (G(c)), and water sorption. Six experimental resins were formulated from dimethacrylate resins, and were filled to 86 wt% (ca. 70 vol%) with treated inorganic filler to form six experimental composites. The fracture toughness was determined using a double torsion technique, the elastic modulus was measured in flexure, and the fracture energy was calculated from the fracture toughness and elastic modulus. As a result of aging in water, K(c) and the G(c) increased, and the elastic modulus decreased, but all values approached a plateau near 6 wk. Water sorption also occurred during this period, mainly during the first 2 wk. Variations in the mechanical properties are interpreted as being due to plasticization of the resin matrix by water, which appears to lower the yield stress and increase in the size of the plastic zone ahead of the crack, thereby causing the observed increase in G(c) and K(c). After approximately 6 wk, no further changes in properties occurred.


A Noonan-like short stature syndrome with sparse hair.
Noonan's syndrome is a clinically recognisable short stature syndrome with autosomal dominant inheritance. The diagnosis can be difficult as the phenotypic expression is very variable. There has been an attempt to divide this syndrome into type I (in which the facial features, especially ptosis, antimongoloid eye slant, and hypertelorism are prominent) and type II (where cardiological abnormalities are more to the fore), but this has not yet been confirmed by other studies.


[ Structure of the gastroduodenal mucosa after surgical treatment of complicated ulcer disease].
There were examined 35 patients after performance of operations for complicated ulcer disease. To the first group were incorporated 9 (25.7%) patients, in whom palliative operations were done, to the second group--9 (25.7%) patients, to whom vagotomy with gastric drainage operation was performed, and to the third group--17 (48.6%) patients, in whom gastric resections were applied. After palliative operations patholologic mucosal changes were localized predominantly in gastric antrum and duodenal ampulla. In 89% of observations mucosal pathology was associated with H. pylori. In 77.8% of the examined patients of second group the mucosal contamination by H. pylori was revealed. The relation was determined between gastric antrum mucosal inflammation and H. pylori infectioning. After gastric resection more significant signs of mucosal inflammation were observed in gastric stump and anastomotic region in comparison with such in efferent intestinal loop. Studying of gastroduodenal mucosa, using histological investigations after surgical treatment of complicated ulcer disease permits to reveal the character of mucosal pathological changes after performance of various operations and, taking it to account, to elaborate the individualized program of the patients rehabilitation.


LncRNA Tug1 involves in the pulmonary vascular remodeling in mice with hypoxic pulmonary hypertension via the microRNA-374c-mediated Foxc1.
Hypoxic pulmonary hypertension (HPH) is a serious and potentially devastating disorder of the pulmonary circulation with complicated mechanisms. Long non-coding RNA (lncRNA) has been revealed to participate in HPH development. This study aimed to explore how lncRNA Tug1 affected the pulmonary vascular remodeling in HPH. A mouse model of HPH and a pulmonary artery smooth muscle cell (PASMC) model of HPH (HPH-PASMCs) were established, where the expression of lncRNA Tug1 was determined. Then, the interaction among lncRNA Tug1, miR-374c, and Foxc1 was assessed. Finally, in order to determine the effects of lncRNA Tug1 on PASMC activities and pulmonary vascular remodeling after HPH, the expression of lncRNA Tug1 was silenced in HPH-PASMCs and HPH mice, with the proliferation, apoptosis, and migration of PASMCs as well as blood pressure in mice measured, respectively. The obtained data revealed that lncRNA Tug1 was highly expressed in HPH mice and HPH-PASMCs. In addition, lncRNA Tug1 up-regulated the expression of Foxc1 by binding to miR-374c. Notably, silencing of lncRNA Tug1 inhibited the proliferation and migration, but promoted the apoptosis of PASMCs. Moreover, lncRNA Tug1 silencing was observed to attenuate the pulmonary vascular remodeling in HPH mice through the Foxc1-mediated NOTCH signaling pathway. Taken conjointly, silencing of lncRNA Tug1 down-regulated the Foxc1 expression by binding to miR-374c, thereby inhibiting the proliferation and migration, while promoting apoptosis of PASMCs to impede pulmonary vascular remodeling in HPH via the Notch signaling pathway. This study provided novel therapeutic insights for treating HPH.


Replantation of a transmetacarpal amputation. A case report.
A case is presented of a 23-year-old male who sustained a traumatic transmetacarpal amputation of his nondominant hand. The injury consisted of complete severance of structures distal to the midpalm. Microsurgical reconstruction involved the primary repair of arteries, veins, nerves, extensor and flexor tendons, and metacarpal fractures. Skeletal reconstruction also employed a primary Swanson prosthesis for the fifth metacarpophalangeal (MP) joint. Early postoperative range-of-motion exercises were encouraged, with the achievement of a functionally capable replanted extremity. The general management of an amputation injury is also discussed, as it applies to a community hospital environment.


Differences in Bcl-2 expression by T-cell subsets alter their balance after in vivo irradiation to favor CD4+Bcl-2hi NKT cells.
Although it is well known that in vivo radiation depletes immune cells via the Bcl-2 apoptotic pathway, a more nuanced analysis of the changes in the balance of immune-cell subsets is needed to understand the impact of radiation on immune function. We show the balance of T-cell subsets changes after increasing single doses of total body irradiation (TBI) or after fractionated irradiation of the lymphoid tissues (TLI) of mice due to differences in radioresistance and Bcl-2 expression of the NKT-cell and non-NKT subsets to favor CD4(+)Bcl-2(hi) NKT cells. Reduction of the Bcl-2(lo) mature T-cell subsets was at least 100-fold greater than that of the Bcl-2(hi) subsets. CD4(+) NKT cells upregulated Bcl-2 after TBI and TLI and developed a Th2 bias after TLI, whereas non-NKT cells failed to do so. Our previous studies showed TLI protects against graft versus host disease in wild-type, but not in NKT-cell-deficient mice. The present study shows that NKT cells have a protective function even after TBI, and these cells are tenfold more abundant after an equal dose of TLI. In conclusion, differential expression of Bcl-2 contributes to the changes in T-cell subsets and immune function after irradiation.


The rules for symbiont community assembly change along a mutualism-parasitism continuum.
Symbiont community assembly is driven by host-symbiont and symbiont-symbiont interactions. The effects that symbionts exert on their hosts are often context-dependent, and existing theoretical frameworks of symbiont community assembly do not consider the implications of variable outcomes to assembly processes. We hypothesized that symbiont-symbiont interactions become increasingly important along a parasitism/mutualism continuum because; (i) negative outcomes favour host resistance which in turn reduces symbiont colonization and subsequently reduce symbiont-symbiont interactions, whereas (ii) positive host outcomes favour tolerance and consequently higher symbiont colonization rates, leading to stronger interactions among symbionts. We found support for this hypothesis in the cleaning symbiosis between crayfish and ectosymbiotic branchiobdellidan worms. The symbiosis between crayfish and their worms can shift from parasitism/commensalism to mutualism as crayfish age. Here, field surveys identified changes in worm density, diversity and composition that were concomitant to changing symbiosis outcomes. We conducted several laboratory experiments and behavioural assays to relate patterns from the field to their likely causal processes. Young crayfish typically hosted only two relatively small worm species. Older crayfish hosted two additional larger species. In laboratory experiments, young crayfish exhibited a directed grooming response to all worm species, but were unable to remove small species. Conversely, adult crayfish did not exhibit grooming responses to any worm species. Relaxed grooming allowed the colonization of large worm species and initiated symbiont-symbiont intraguild predation that reduced the abundance and altered the behaviour of small worm species. Thus, the dominant processes of symbiont community assembly shifted from host resistance to symbiont-symbiont interactions through host ontogeny and a concomitant transition towards mutualism. This work shows that host resistance can have a prevailing influence over symbiont community assembly when symbiosis is disadvantageous to the host. However, when symbiosis is advantageous and resistance is relaxed, symbiont colonization rate and consequently abundance and diversity increases and interactions among symbionts become increasingly important to symbiont community assembly.


Human bone marrow stromal cell colonies: response to hydrocortisone and dependence on platelet-derived growth factor.
Long-term bone marrow cultures are dependent on the formation in vitro of an adherent cell layer that supports hematopoiesis. We have grown bone-marrow-adherent cells, termed stromal colony-forming units, or CFU-ST, as isolated adherent colonies, and examined some of their growth requirements. Bone marrow mononuclear cells separated from aspirates by density centrifugation and cultured in medium supplemented with fetal calf serum or human plasma gave rise to adherent colonies (CFU-ST). An average of 23.4 +/- 2.1 (mean +/- SEM, n = 19) CFU-ST were produced by 10(5) bone marrow mononuclear cells. CFU-ST could not be cultured from similarly prepared peripheral blood mononuclear cells. The colonies were composed of spindle cells, flat cells, and fat-containing cells, with all three types often present in the same colony, suggesting derivation from a common progenitor. Cells were negative for nonspecific esterase and factor VIII antigen. Hydrocortisone added to the cultures at concentrations of 10(-7) M induced the formation of adipose cells in the center of one-third to one-half of the colonies but did not affect CFU-ST number. Human platelet-poor plasma and platelet-rich plasma were substituted for fetal calf serum in the medium. When all determinations for four experiments were averaged, platelet-rich plasma gave 17.8 +/- 1.2 (mean +/- SEM, n = 16) colonies, whereas platelet-poor plasma gave only 0.2 +/- 0.1 colonies (n = 15). When purified platelet-derived growth factor (PDGF) was added to platelet-poor plasma, growth of CFU-ST was enhanced, and a dose-response relationship was found between size of colonies and concentration of added PDGF. Granulocyte-macrophage colony stimulating factor added to cultures had no effect on the growth of CFU-ST.


Sequence hydropathy dominates membrane protein response to detergent solubilization.
The ability to predict from amino acid sequence how membrane protein structures will respond to detergent solubilization would significantly facilitate experimental characterization of these molecules. Here we have investigated and compared the response to solubilization by the "mild" n-dodecyl-β-D-maltoside (DDM) and "harsh" sodium dodecyl sulfate (SDS) of wild-type and point mutant "hairpin" (helix-loop-helix) membrane proteins derived from the third and fourth TM segments of the human cystic fibrosis transmembrane conductance regulator (CFTR) and the intervening extracellular loop. Circular dichroism spectroscopy, size-exclusion chromatography, and pyrene fluorescence spectroscopy were used to evaluate the secondary structures, hairpin-detergent complex excluded volumes, and hairpin compactness of the detergent-solubilized sequences. Sequence hydrophobicity is found to be the dominant factor dictating membrane protein response to detergent solubilization by DDM and SDS, with hairpin secondary structure exquisitely sensitive to mutation when DDM is used for solubilization. DDM and SDS differ principally in their ability to promote approach of TM segment ends, although hairpin compactness remains sensitive to point mutations. Our overall findings suggest that protein-protein and protein-detergent interactions are determined concomitantly, with the net hydropathy of residues exposed to detergent dominating the observed properties of the solubilized protein.


Formative assessment and academic achievement in pre-graduate students of health sciences.
Although educational experts recommend the use of formative assessment, there is a dearth of empirical studies on its impact on academic achievement. In this research the authors analyse to what extent participation and performance in formative assessment are associated with positive academic outcomes of pre-graduate students of health sciences. A total of 548 students from three health science degrees (Medicine, Psychology and Biology) from four Spanish universities were involved in this study. The students who carried out mid-term formative assessment got better marks and had higher success rates in final summative assessment that the students who did not participate. In addition, success in formative assessment tests was associated with better summative marks. Interestingly, participation in formative assessment was a better predictor of final outcome than success in formative assessment, a result that supports the key role of feedback in formative assessment. Students who took the mid-term examination, irrespective of their success, obtained feedback about their achievement and probably this determined their greater involvement in the learning process. Although causal relationships between formative and summative assessment cannot be established from this research, the generalized benefits of formative assessments found here encourage the practice of them in health sciences education.


Metabolic and non-metabolic liver zonation is established non-synchronously and requires sinusoidal Wnts.
The distribution of complementary metabolic functions in hepatocytes along a portocentral axis is called liver zonation. Endothelial secreted Wnt ligands maintain metabolic zonation in the adult murine liver but whether those ligands are necessary to initiate zonation in the immature liver has been only partially explored. Also, numerous non-metabolic proteins display zonated expression in the adult liver but it is not entirely clear if their localization requires endothelial Wnts. Here we used a novel transgenic mouse model to compare the spatial distribution of zonated non-metabolic proteins with that of typical zonated metabolic enzymes during liver maturation and after acute injury induced by carbon tetrachloride (CCl4). We also investigated how preventing Wnt ligand secretion from endothelial cells affects zonation patterns under homeostasis and after acute injury. Our study demonstrates that metabolic and non-metabolic zonation are established non-synchronously during maturation and regeneration and require multiple endothelial Wnt sources.


Time to detection of Mycobacterium tuberculosis in sputum culture correlates with outcome in patients receiving treatment for pulmonary tuberculosis.
The purpose of this study was to determine whether the time to detection (TTD) of Mycobacterium tuberculosis in sputum culture correlates with the response to antituberculous treatment in patients with pulmonary tuberculosis. Twenty-six consecutive patients were studied who had active pulmonary tuberculosis and sufficient sputum cultures and clinical follow-up to allow adequate assessment. Following initiation of antituberculous therapy, 13 patients (group 1, responders) had a complete response to treatment, and the TTD of M tuberculosis using the mycobacterial growth indicator tube increased steadily. The remaining 13 patients (group 2, nonresponders) had persistent evidence of active disease and demonstrated little or no increase in the TTD with treatment unless an additional therapeutic intervention was implemented (surgery, improved compliance with medications, or a change in medications). The presence of HIV infection, intravenous drug use, multidrug resistance, treatment with second-line therapy, extensive radiographic involvement, and cavitary disease were associated with a delayed increase in the TTD. The TTD was superior to clinical, radiographic, or conventional bacteriologic evaluation in determining treatment outcome. The TTD closely correlates with the overall response to treatment for pulmonary tuberculosis and may represent a useful adjunct to predict outcome in these patients.


Water extraction and root traits in Oryza sativa × Oryza glaberrima introgression lines under different soil moisture regimes.
Drought is a major constraint to rainfed rice production in some parts of sub Saharan Africa (SSA) because of irregular and low rainfall. Improving root water uptake during progressive soil drying could contribute to terminal drought stress resistance. Two lysimeter experiments were conducted using 200 introgression lines (ILs) derived from Oryza sativa L.×Oryza glaberrima Steud. crosses (IR64/RAM54 and IR64/RAM90) to characterise water uptake and root traits under progressive soil drying and to assess genetic differences among these lines in relation to drought stress. The lysimeter facility setup and protocols for simultaneous monitoring of water uptake rates and leaf area are described. Significant genetic variability for water uptake and root traits among the ILs was observed under drought stress. Water uptake in the drought stress treatments was correlated with root length density (RLD) and root dry matter (RDM) at lower soil depths. The highest RLD values at depth were observed among the RAM90 ILs and genetic differences were observed between RAM90 and RAM54 populations, implying that accession RAM90 could be used as a source for improving drought avoidance in lowland rice. The ILs with greater water uptake identified in this study will be used in future research for improving drought resistance of rice in SSA.


PedSplit: pedigree management for stratified analysis.
PedSplit facilitates pedigree management for gene-gene interaction and sex specific tests for increased homogeneity within subgroups. PedSplit also provides a simple approach for calculating haplotype relative risk and generating internal 'controls'. Executables, C++ source code and documentation for PedSplit can be downloaded from http://www.pharmacogenetics.ca in the links and software section.


Biochemical compartmentation of fish tissues. Hexokinase in the brain.
Differential distribution of gross, particulate and soluble forms of hexokinase in the cerebrum, cerebellum, pituitary, hypothalamus and thalamus and medulla oblongata was studied in 9 inland teleosts of different phylogenetic age to clarify the compartmentation of glucose metabolism. Enzyme activity (gross) was maximum in the cerebrum and minimum in the medulla oblongata, suggesting that glucose metabolism is more intensive in the former structure. The activity in the particulate fraction was highest in the cerebellum followed by the cerebrum, midbrain and medulla oblongata. Gross, particulate and soluble fraction of the enzyme was higher in major carp species than in cat fish or snake head species. Total particular and soluble fraction was highest in the cerebrum, cerebellum and midbrain. The distributional trend of the enzyme was similar in the 9 fishes.


Preconditioning stimuli induce autophagy via sphingosine kinase 2 in mouse cortical neurons.
Sphingosine kinase 2 (SPK2) and autophagy are both involved in brain preconditioning, but whether preconditioning-induced SPK2 up-regulation and autophagy activation are linked mechanistically remains to be elucidated. In this study, we used in vitro and in vivo models to explore the role of SPK2-mediated autophagy in isoflurane and hypoxic preconditioning. In primary mouse cortical neurons, both isoflurane and hypoxic preconditioning induced autophagy. Isoflurane and hypoxic preconditioning protected against subsequent oxygen glucose deprivation or glutamate injury, whereas pretreatment with autophagy inhibitors (3-methyladenine or KU55933) abolished preconditioning-induced tolerance. Pretreatment with SPK2 inhibitors (ABC294640 and SKI-II) or SPK2 knockdown prevented preconditioning-induced autophagy. Isoflurane also induced autophagy in mouse in vivo as shown by Western blots for LC3 and p62, LC3 immunostaining, and electron microscopy. Isoflurane-induced autophagy in mice lacking the SPK1 isoform (SPK1(-/-)), but not in SPK2(-/-)mice. Sphingosine 1-phosphate and the sphingosine 1-phosphate receptor agonist FTY720 did not protect against oxygen glucose deprivation in cultured neurons and did not alter the expression of LC3 and p62, suggesting that SPK2-mediated autophagy and protections are not S1P-dependent. Beclin 1 knockdown abolished preconditioning-induced autophagy, and SPK2 inhibitors abolished isoflurane-induced disruption of the Beclin 1/Bcl-2 association. These results strongly indicate that autophagy is involved in isoflurane preconditioning both in vivo and in vitro and that SPK2 contributes to preconditioning-induced autophagy, possibly by disrupting the Beclin 1/Bcl-2 interaction.


Epidemiological and clinical characteristics of the chikungunya outbreak in Ruili City, Yunnan Province, China.
Chikungunya fever is an acute infectious disease caused by the chikungunya virus (CHIKV) that is characterized by fever, rash, and joint pain. CHIKV has infected millions of people in Africa, Asia, America, and Europe since it re-emerged in the Indian Ocean region in 2004. Here, we report an outbreak of Chikungunya fever that occurred in Ruili of Yunnan Province, a city located on the border between China and Myanmar, in September 2019. The outbreak lasted for three months from September to December. Overall, 112 cases were confirmed by a real-time reverse-transcription polymerase chain reaction in the Ruili People's Hospital, and they showed apparent temporal, spatial, and population aggregation. Among them, 91 were local cases distributed in 19 communities of Ruili City, and 21 were imported cases. The number of female patients was higher than that of male patients, and most patients were between 20 and 60 years old. The main clinical manifestations included joint pain (91.96%), fever (86.61%), fatigue (58.04%), chills (57.14%), rash (48.21%), headache (39.29%), and so forth. Biochemical indexes revealed increased C-reactive protein (63.39%), lymphopenia (57.17%), increased hemoglobin (33.04%), neutrophilia (28.57%), and thrombocytopenia (16.07%). Phylogenetic analysis of the complete sequences indicated that the CHIKV strains in this outbreak belonged to the Indian Ocean clade of the East/Central/South African genotype. We speculated that this chikungunya outbreak might be caused by CHIKV-infected persons returning from Myanmar, and provided a reference for the formulation of effective treatment and prevention measures.


Coevolution of white matter hyperintensities and cognition in the elderly.
To investigate the effects of baseline white matter hyperintensity (WMH) and rates of WMH extension and emergence on rate of change in cognition (episodic memory and executive function). A total of 150 individuals including cognitively normal elderly individuals and those with Alzheimer disease and mild cognitive impairment completed serial episodic memory and executive function evaluations and serial MRI scans sufficient for longitudinal measurement of WMH (mean delay 4.0 years). Incident WMH voxels were categorized as extended (baseline WMH that grew larger) or emergent (newly formed WMH). We used a stepwise regression approach to investigate the effects of baseline WMH and rates of WMH extension and emergence on rate of change in cognition (episodic memory and executive function). WMH burden significantly increased over time, and approximately 80% of incident WMH voxels represented extensions of existing lesions. Each 1 mL/y increase in WMH extension was associated with an additional 0.70 SD/y of subsequent episodic memory decrease (p = 0.0053) and an additional 0.55 SD/y of subsequent executive function decrease (p = 0.022). Emergent WMHs were not found to be associated with a change in cognitive measures. Aging-associated WMHs evolve significantly over a 4-year period. Most of this evolution represents worsening injury to the already compromised surround of existing lesions. Increasing WMH was also significantly associated with declining episodic memory and executive function. This finding supports the view that white matter disease is an insidious and continuously evolving process whose progression has clinically relevant cognitive consequences.


3,4-Ethylenedioxythiophene (EDOT) End-Group Functionalized Poly-ε-caprolactone (PCL): Self-Assembly in Organic Solvents and Its Coincidentally Observed Peculiar Behavior in Thin Film and Protonated Media.
End-group functionalization of homopolymers is a valuable way to produce high-fidelity nanostructured and functional soft materials when the structures obtained have the capacity for self-assembly (SA) encoded in their structural details. Herein, an end-functionalized PCL with a π-conjugated EDOT moiety, (EDOT-PCL), designed exclusively from hydrophobic domains, as a functional "hydrophobic amphiphile", was synthesized in the bulk ROP of ε-caprolactone. The experimental results obtained by spectroscopic methods, including NMR, UV-vis, and fluorescence, using DLS and by AFM, confirm that in solvents with extremely different polarities (chloroform and acetonitrile), EDOT-PCL presents an interaction- and structure-based bias, which is strong and selective enough to exert control over supramolecular packing, both in dispersions and in the film state. This leads to the diversity of SA structures, including spheroidal, straight, and helical rods, as well as orthorhombic single crystals, with solvent-dependent shapes and sizes, confirming that EDOT-PCL behaves as a "block-molecule". According to the results from AFM imaging, an unexpected transformation of micelle-type nanostructures into single 2D lamellar crystals, through breakout crystallization, took place by simple acetonitrile evaporation during the formation of the film on the mica support at room temperature. Moreover, EDOT-PCL's propensity for spontaneous oxidant-free oligomerization in acidic media was proposed as a presumptive answer for the unexpected appearance of blue color during its dissolution in CDCl3 at a high concentration. FT-IR, UV-vis, and fluorescence techniques were used to support this claim. Besides being intriguing and unforeseen, the experimental findings concerning EDOT-PCL have raised new and interesting questions that deserve to be addressed in future research.


[Long Noncoding RNAs MEG3, TUG1, and hsa-miR-21-3p Are Potential Diagnostic Biomarkers for Coronary Artery Disease].
Peripheral blood biomarkers are of particular importance to diagnose certain diseases including coronary artery disease (CAD) due to their non-invasiveness. Investigating the expression of noncoding RNAs (ncRNAs) paves the way to early disease diagnosis, prognosis, and treatment. Consequently, in this research, we aimed to investigate a panel of ncRNAs as potential biomarkers in patients with coronary artery disease. Two different groups have been designed (control and CAD). All participants were subjected to interviews and clinical examinations. Peripheral blood samples were collected, and plasma was extracted. At the same time, target ncRNAs have been selected based on literature review and bioinformatic analysis, and later they underwent investigation using quantitative real-time PCR. The selected panel encompassed the long non-coding RNAs (lncRNAs) MEG3, TUG1, and SRA1, and one related microRNA (miRNA): hsa-miR-21-3p. We observed statistically significant upregulation in MEG3, TUG1, and hsa-miR21-3p in CAD patients compared to control participants (p-value < 0.01). Nevertheless, SRA1 exhibited downregulation with no statistical significance (p-value > 0.05). All ncRNAs under study displayed a significantly strong correlation with disease incidence, age, and smoking. Network construction revealed a strong relationship between MEG3 and TUG1. ROC analysis indicated high potentiality for hsa-miR-21-3p to be a promising biomarker for CAD. Moreover, MEG3 and TUG1 displayed distinguished diagnostic discrimination but less than hsa-miR-21-3p, all of them exhibited strong statistical significance differences between CAD and control groups. Conclusively, this research pinpointed that MEG3, TUG1, and hsa-miR-21-3p are potential biomarkers of CAD incidence and diagnosis.


Factors Hindering Practice of Day Care Surgery in a Tertiary Care Centre in Southern India: A Patient's Perspective.
Day care surgery offers cost containment, effective usage of hospital beds, reduced incidence of nosocomial infection and early recovery in home environment. In developing countries like India, there are various factors that influence the success of day care surgery. To assess the factors hindering the practice of day care surgery in a tertiary care centre in South India. This observational study was conducted in the Department of General Surgery, JIPMER, Puducherry, India, from January 2013 to March 2014. All male patients with uncomplicated inguinal hernia who were admitted for elective surgery under one particular surgery unit and who were found fit for discharge on Postoperative Day 1 (POD1) based on clinical fitness were included in the study. A questionnaire containing the patient's acceptance decision, VAS (Visual Analogue Scale) pain score and the reason for non acceptance if any was used for assessment. Among the 89 patients who were fit for discharge on POD1, the decision for discharge was accepted by 57 patients. 32 patients were not satisfied of the decision for discharge on POD1. The common reasons for dissatisfaction with the decision were persistent pain at operated site (13 patients with mean VAS score 8.3), non availability of health care resources in their locality (12 patients) and unwillingness to travel on POD1 (four patients). A comprehensive and well presented preoperative counselling along with an effective primary health service would help in promoting day care surgery in developing countries.


Biological effects of air pollution on the function of human skin equivalents.
The World Health Organization reports that 99% of the global population are exposed to pollution levels higher than the recommended air quality guidelines. Pollution-induced changes in the skin have begun to surface; however, the effects require further investigation so that effective protective strategies can be developed. This study aimed to investigate some of the aging-associated effects caused by ozone and particulate matter (PM) on human skin equivalents. Full-thickness skin equivalents were exposed to 0.01 μg/μL PM, 0.05 μg/μL PM, 0.3 ppm ozone, or a combination of 0.01 μg/μL PM and 0.3 ppm ozone, before skin equivalents and culture medium were harvested for histological/immunohistochemical staining, gene and protein expression analysis using qPCR, Western blotting, and ELISA. Markers include MMP-1, MMP-3, COL1A1, collagen-I, 4-HNE, HMGCR, and PGE2. PM was observed to induce a decrease in epidermal thickness and an enhanced matrix building phenotype, with increases in COL1A1 and an increase in collagen-I protein expression. By contrast, ozone induced an increase in epidermal thickness and was found to induce a matrix-degrading phenotype, with decreases in collagen-I gene/protein expression and increases in MMP-1 and MMP-3 gene/protein expression. Ozone was also found to induce changes in lipid homeostasis and inflammation induction. Some synergistic damage was also observed when combining ozone and 0.01 μg/μL PM. The results presented in this study identify distinct pollutant-induced effects and show how pollutants may act synergistically to augment damage; given individuals are rarely only exposed to one pollutant type, exposure to multiple pollutant types should be considered to develop effective protective interventions.


Fetal cystic hygroma and Turner's syndrome.
Large nuchal cystic hygromas were observed in five second-trimester aborted fetuses at autopsy. Two female fetuses with generalized edema were karyotyped as 45,X. One of these was the twin of a 46,XX normal female sibling. The association of generalized edema with large nuchal cystic hygromas was seen only in these two fetuses and represents strong phenotypic evidence of Turner's syndrome. However, the absence of hydrops was not a reliable indicator of normal karyotype. One fetus without generalized edema was karyotyped as 47,XY, +21, inv(9). The remaining cases had normal karyotypes. Placental histology was not useful in discriminating monosomy X from other conditions, but placental tissue culture was important in obtaining a cytogenetic diagnosis. Karyotyping is recommended in all cases of fetal cystic hygroma.


Hannaella siamensis sp. nov. and Hannaella phetchabunensis sp. nov., two new anamorphic basidiomycetous yeast species isolated from plants.
Eight strains, representing two novel anamorphic yeast species, consisted of five strains isolated from the external surfaces of rice leaves (DMKU-RP72(T), DMKU-RP109, DMKU-RP119, YE-124 and YE-156) and one from a corn leaf (DMKU-CP430(T))4 collected in Thailand, and one strain isolated from each of a composite flower (11-1114) and a fallen dead leaf (12-301); the latter two were collected in Belize. On the basis of sequence analysis of the D1/D2 region of the large subunit rRNA gene and the internal transcribed spacer (ITS) region, they were suggested to be two novel species of the genus Hannaella. Seven strains (DMKU-RP72(T), DMKU-RP109, DMKU-RP119, YE-124, YE-156, 11-1114 and 12-301) differed from each other by 0-3 nt substitutions in the D1/D2 region and by 0-1 nt substitutions in the ITS region. In terms of pairwise sequence similarities of the D1/D2 region these seven strains were closest to Hannaella zeae, but with 1.2-1.7% (7-9) nucleotide substitutions. The sequences of the ITS region of these seven strains differed from H. zeae by 3.7-3.9% (16-17) nucleotide substitutions. Therefore, they were assigned to a single novel species and the name Hannaella siamensis sp. nov. has been proposed. The type strain is DMKU-RP72(T) ( = BCC 69493(T) = NBRC 110425(T) = CBS 13533(T)). Strain DMKU-CP430(T) represents the second novel species and was also most closely related to H. zeae, but with 1.0% (6) nucleotide substitutions in the D1/D2 region and 3.2% (14) nucleotide substitutions in the ITS region. It was assigned to the proposed novel species, Hannaella phetchabunensis sp. nov. (type strain DMKU-CP430(T) = BCC 69492(T) = NBRC 110424(T) = CBS 13386(T)).


The development of the antitumour benzothiazole prodrug, Phortress, as a clinical candidate.
This review traces the development of a series of potent and selective antitumour benzothiazoles from the discovery of the initial lead compound, 2-(4-amino-3-methylphenyl)benzothiazole (DF 203) in 1995 to the identification of a clinical candidate, Phortress, scheduled to enter Phase 1 trials in Q1 2004 under the auspices of Cancer Research U.K. Advances in our understanding of the mechanism of action of this unique series of agents are described and can be summarised as follows: selective uptake into sensitive cells followed by Arylhydrocarbon Receptor (AhR) binding and translocation into the nucleus, induction of the cytochrome P450 isoform (CYP) 1A1, conversion of the drug into an electrophilic reactive intermediate and formation of extensive DNA adducts resulting in cell death. Our understanding of this mechanistic scenario has played a crucial role in the drug development process, most notably in the synthesis of fluorinated DF 203 analogues to thwart deactivating oxidative metabolism (5F 203) and water-soluble prodrug design for parenteral administration. Aspects of mechanism of action studies, in vitro and in vivo screening, synthetic chemistry and pharmacokinetics are reviewed here.


How to treat the untreated: effectiveness of a self-help metacognitive training program (myMCT) for obsessive-compulsive disorder.
Despite advances in the understanding and treatment of obsessive-compulsive disorder (OCD), many patients undergoing interventions display incomplete symptom reduction, Our research group has developed a self-help manual entitled "My Metacognitive Training for OCD" (myMCT) aimed at raising patients' awareness about cognitive biases that seem to subserve OCD. The training is particularly intended for patients currently unable or unwilling to attend standard therapy, or in cases where such a treatment option is not available. For the present study, 86 individuals suffering from OCD were recruited over the Internet. Following the initial assessment, participants were either immediately emailed the myMCT manual or allocated to a waitlist group. After 4 weeks, a second assessment was performed, The myMCT group showed significantly greater improvement for OCD symptoms according to the Y-BOCS total score compared with the waitlist group (d = .63), particularly for obsessions (d= .69). Medium to strong differences emerged for the OCI-R (d = .70) and the BDI-SF (d = .50). The investigation provides the first evidence for the effectiveness of the myMCT for OCD.


The Grass Carp Genomic Visualization Database (GCGVD): an informational platform for genome biology of grass carp.
With the release of the draft genome of the grass carp, researches on the grass carp from the genetic level and the further molecular mechanisms of economically valuable physiological behaviors have gained great attention. In this paper, we integrated a large number of genomic, genetic and some other data resources and established a web-based grass carp genomic visualization database (GCGVD). To view these data more effectively, we visualized grass carp and zebrafish gene collinearity and genetic linkage map using Scalable Vector Graphics (SVG) format in the browser, and genomic annotations by JBrowse. Furthermore, we carried out some preliminary study on a whole-genome alternative splicing (AS)of the grass carp. The RNA-seq reads of 15 samples were aligned to the reference genome of the grass carp by Bowtie2 software. RNA-seq reads of each sample and density map of reads were also exhibited in JBrowse. Additionally, we designed a universal grass carp genome annotation data model to improve the retrieval speed and scalability. Compared with the published database GCGD previously, we newly added the visualization of some more genomic annotations, conserved domain and RNA-seq reads aligned to the reference genome. GCGVD can be accessed at http://220.127.116.11.


The art and design of genetic screens: caenorhabditis elegans.
The nematode Caenorhabditis elegans was chosen as a model genetic organism because its attributes, chiefly its hermaphroditic lifestyle and rapid generation time, make it suitable for the isolation and characterization of genetic mutants. The most important challenge for the geneticist is to design a genetic screen that will identify mutations that specifically disrupt the biological process of interest. Since 1974, when Sydney Brenner published his pioneering genetic screen, researchers have developed increasingly powerful methods for identifying genes and genetic pathways in C. elegans.


Combined liver-kidney transplants: allosensitization and recipient outcomes.
A pretransplant positive crossmatch in combined liver kidney transplants (CLK) is not considered a contraindication based on the reported immunoprotection conferred by the liver allograft. However, antibody-mediated rejection of the kidney in CLK has been reported recently. This prompted our study to investigate the impact of presensitization on CLK recipient outcomes. We examined kidney allograft and patient survival by indication of sensitization using Scientific Registry of Transplant Recipients data on CLK performed from 1995 to 2008. We defined sensitization as panel reactive antibody (PRA) more than 10% or a positive T-cell crossmatch (TXM). Among 2484 CLK recipients with available PRA or TXM information, 30% had positive TXM or PRA more than 10%. Among those with TXM information, 12% had a positive crossmatch (n=234). In univariate analyses, patient (P=0.002) and overall kidney graft survival (P=0.015) were significantly diminished among sensitized patients. Differences in patient survival translated to estimated half-lives of 10.3 years among nonsensitized recipients versus 7.8 years among sensitized recipients, In multivariable Cox models, allosensitization was independently associated with patient death (adjusted hazard ratio=1.22, 95% CI, 1.04-1.43) and overall kidney graft loss (adjusted hazard ratio=1.16, 95% CI, 1.00-1.36). These results suggest a negative impact of presensitization on patient and overall renal allograft survival in CLK. Accordingly, presensitization may need to be considered in risk stratification and clinical management of CLK.


Emodin ameliorates tubulointerstitial fibrosis in obstructed kidneys by inhibiting EZH2.
Emodin, a major component of Chinese herbal rhubarb, delays the progression of chronic renal failure. However, the effect and working mechanisms of Emodin on renal tubulointerstitial fibrosis remains elusive. We hypothesized that emodin inhibits renal tubulointerstitial fibrosis through EZH2, a histone methyltransferase. Our in vivo and in vitro studies demonstrate that emodin reduced extracellular collagen deposition and inhibited Smad3 and CTGF pro-fibrotic signaling pathways, which were correlated with the down-regulation of EZH2 and reduced trimethylation of histone H3 on lysine 27 (H3k27me3) in NRK-49F fibrotic cells and UUO kidneys. Inhibition of EZH2 by 3-DZNeP blocked or attenuated the anti-fibrotic effect of emodin in UUO kidneys and NRK-49F cells. These data indicate that emodin inhibits renal tubulointerstitial fibrosis in obstructed kidneys and this effect is mediated through EZH2.


Brain-on-a-Chip: A Human 3D Model for Clinical Application.
The main goal of this research is to design, develop and implement an efficient protocol to generate 3D neural cultures derived from human induced Pluripotent Stem Cells (hiPSCs) coupled to Micro Electrode Arrays (MEA) in order to obtain an engineered and controlled brain-on-a-chip model. The use of patient specific iPSCs may offer novel insights into the pathophysiology of a large variety of disorders, including numerous neurodevelopmental and late-onset neurodegenerative conditions. With these in vitro patient specific models, we may have the possibility to test drugs and find ad hoc therapies in the direction of precision medicine.


Tip-localized Ca2+ -permeable channels control pollen tube growth via kinase-dependent R- and S-type anion channel regulation.
Pollen tubes (PTs) are characterized by having tip-focused cytosolic calcium ion (Ca2+ ) concentration ([Ca2+ ]cyt ) gradients, which are believed to control PT growth. However, the mechanisms by which the apical [Ca2+ ]cyt orchestrates PT growth are not well understood. Here, we aimed to identify these mechanisms by combining reverse genetics, cell biology, electrophysiology, and live-cell Ca2+ and anion imaging. We triggered Ca2+ -channel activation by applying hyperpolarizing voltage pulses and observed that the evoked [Ca2+ ]cyt increases were paralleled by high anion channel activity and a decrease in the cytosolic anion concentration at the PT tip. We confirmed a functional correlation between these patterns by showing that inhibition of Ca2+ -permeable channels eliminated the [Ca2+ ]cyt increase, resulting in the abrogation of anion channel activity via Ca2+ -dependent protein kinases (CPKs). Functional characterization of CPK and anion-channel mutants revealed a CPK2/20/6-dependent activation of SLAH3 and ALMT12/13/14 anion channels. The impaired growth phenotypes of anion channel and CPK mutants support the physiological significance of a kinase- and Ca2+ -dependent pathway to control PT growth via anion channel activation. Other than unveiling this functional link, our membrane hyperpolarization method allows for unprecedented manipulation of the [Ca2+ ]cyt gradient or oscillations in the PT tips and opens an array of opportunities for channel screenings.


[Polyspermism and antipodal fertilization in Lilium (Tourn.) L. species].
It has been shown that several pollen tubes can penetrate into the embryo sac on the source side of the antipodal apparatus. One of the pair of sperms of additional pollen tube copulates with the upper antipodal, the second sperm copulates with the lower antipodal or rarely penetrates in the central cell. The process of fertilization was accomplished by the phase of nuclei morphological similarity characteristic ofsyngamy (by postmitotic type of fertilization according to Gerasimova-Navashina). A directional growth of additional pollen tubes involves a specifically differentiation of antipodal cells that imitates the egg cell.


Sulfated polysaccharides of brown seaweed Cystoseira canariensis bind to serum myostatin protein.
Natural sulfated polysaccharides (SPs) derived from brown seaweed comprise a complex group of macromolecules with a wide range of important physiological properties. SPs have been shown to bind and directly regulate the bioactivity of growth factors and cytokines such as basic fibroblast growth factor, interferon, various enzymes and transforming growth factor. Myostatin is a member of the transforming growth factor-beta (TGF-beta) family that acts as a negative regulator of skeletal muscle mass. In this work we demonstrated that SPs isolated from the brown seaweed Cystoseira canariensis bind to the myostatin protein in serum.


Synthesis of benzophenone-containing analogues of phosphatidylcholine.
As part of a collaborative study of cellular efflux of cholesterol and phospholipids, photoactivable analogues 4-8 of phosphatidylcholine (PC) having benzophenone groups in the choline moiety and at the end of the C2 and C1 alkyl chains have been synthesized. The efficient preparation via Suzuki coupling of the appropriate long-chain benzophenone-containing carboxylic acid and alcohol and their incorporation by adaptation of known approaches into the acyl- and ether-linked PC analogues 6-8 are described. Development of a method for radiolabeling these PC analogues, via hydrogenation of a double bond in modified side chains, is also described.


Increased prevalence of group A β-hemolytic streptococcus among an ethnic population in Kyrgyzstan detected by the rapid antigen detection test.
The incidence of rheumatic fever (RF) has markedly increased in the last 10 years in Kyrgyzstan. Therefore, investigating the prevalence of group A β-hemolytic streptococcus (GABHS), which is the cause of RF, in the Kyrgyzstan population is crucial. We studied 189 subjects: 59 children [29 with RF and/or rheumatic heart disease (RHD)] and 130 adults (15 with RHD). The average age of the subjects was 41.0±10.0 years (range 8 months to 72 years). A general clinical examination and medical history including eating habits was carried out. The prevalence of GABHS was tested using the highly sensitive rapid antigen detection test (RADT) to detect the outcrop of streptococcus antigen in smears taken from the mucosal surface of the tonsils or the back of the throat. GABHS antigen was positive in 70 of a total 189 subjects [37.0%; 22/59 children (37.2%), 48/130 adults (36.9%)]. In patients with RF/RHD (n=44), GABHS was positive in 14 subjects [31.8%; 8/29 children (27.6%), 6/15 adults (40.0%)]. Thirty-two subjects with RF/RHD had frequent episodes of tonsillopharyngitis. In subjects without RF/RHD (n=145), GABHS was positive in 56 subjects [38.6%; 14/30 children (46.6%), 42/115 adults (36.5%)]. Thirty of these subjects had frequent episodes of tonsillopharyngitis. Of the 130 adults, the most-consumed dairy products included yoghurt (n=115; 88.4%), milk kasha (n=75; 57.7%) and milk (n=40; 30.7%). Of the 115 subjects in the yoghurt-consuming group, 44 (38.2%) had positive results for GABHS. In the non-yoghurt-consuming group, 4/15 subjects (26.6%) had positive results for GABHS. Using RADT for GABHS, a high prevalence of GABHS antigen was detected not only in patients with RF/RHD, but also in the healthy population (without RF/RHD). The low GABHS prevalence in children with RF/RHD (27.6%) was probably due to corresponding antibiotic therapy. In conclusion, the high prevalence of GABHS is one of the main reasons for the rapid increase in RF/RHD in Kyrgyzstan, and RADT would be an effective tool for its detection.


Divergent molecular pathways of productive and latent infection with a virulent strain of herpes simplex virus type 1.
Mutants of herpes simplex virus (HSV) have been used to show that a variety of key genes associated with initiation of lytic infection or replication of viral DNA are not essential for establishment of latency. These observations are extended in the present study, in which a virulent strain of HSV type 1 that is not compromised in its ability to productively infect neurons under favorable conditions was used to demonstrate early divergence of molecular pathways leading to productive and latent infection. Our experimental strategy made unique use of the segmental innervation of the vertebrate trunk to study the spread of virus throughout the peripheral nervous system after inoculation of mouse flanks. Evidence of viral gene expression, including that of immediate-early genes, was transient, confined to ganglia directly innervating the inoculated skin (8th through 12th thoracic segments), and seen only at sites from which infectious virus could be recovered. In contrast, neurons containing latency-associated transcripts and reactivatable virus were more widely distributed (sixth thoracic through first lumbar segments), from which we conclude that replication-competent HSV type 1 can establish latency without initiating productive infection.


Methylmercury inhibits the in vitro uptake of the glutathione precursor, cystine, in astrocytes, but not in neurons.
Maintenance of adequate intracellular glutathione (GSH) levels is vital for intracellular defense against oxidative damage. The toxic effects of methylmercury (MeHg) are attributable, at least in part, to elevated levels of reactive oxygen species, and thus decreases in GSH synthesis may increase methylmercury toxicity. Astrocytes have recently been proposed to play an essential role in providing GSH precursors to neurons. Therefore, cystine transport, a prerequisite to GSH production, was characterized in cultured astrocytes and neurons, and the effects of methylmercury on this transport were assessed. Astrocytes and neurons both possessed temperature dependent transport systems for cystine. Astrocytes accumulated cystine by Na+-independent (X(C)-) and -dependent (X(AG)-) systems while neurons used exclusively Na+-independent systems. Inhibition of the X(AG)- transport system decreased cystine transport in astrocytes to levels equivalent to those in sodium-depleted conditions, suggesting that cystine is carried by a glutamate/aspartate transporter in astrocytes. Inhibition of the multifunction ectoenzyme/amino acid transporter gamma-glutamyltranspeptidase (GGT) decreased cystine transport in both neurons and astrocytes. Inhibition of System X(C)- with quisqualate also decreased cystine uptake in both astrocytes and neurons. These data demonstrate that cultured astrocytes accumulate cystine via three independent mechanisms, System X(AG)-, System X(C)-, and GGT, while cultured hippocampal neurons use System X(C)- and GGT exclusively. Inhibition of cystine uptake in astrocytes by methylmercury appears to be due to actions on the System X(AG)- transporter.


Laminar airflow protection in bone marrow transplantation.
A laminar airflow room was used to provide a low-pathogen environment for a child with lymphopenic immune deficiency transplanted with paternal bone marrow. Comparison of flora from the patient, personnel, and the environment indicated that no colonization with exogenous organisms occurred in the patient during the 45-day period of study. The number of organisms recovered from the laminar airflow room was exceedingly small. Conventional hospital isolation rooms contained more bacteria and fungi than the laminar airflow room, even when strict aseptic procedures were followed in the former. Patients with lymphopenic immune deficiency and agranulocytosis admitted to conventional isolation rooms were colonized with exogenous organisms within 1 week. Each developed infection with these strains, and one patient died. Laminar airflow isolation seems at present the best means to prevent exogenous infection during hospitalization of patients with lymphopenic and other severe immune-deficiency diseases and may be essential when bone marrow transplantation is performed to treat their immunological defect.


Crotalaria-induced pulmonary hypertension. Uptake of 3H-thymidine by the cells of the pulmonary circulation and alveolar walls.
Feeding with Crotalaria spectabilis seeds induces structural changes in the pulmonary arterial circulation characteristic of pulmonary hypertension: increased medial and adventitial thickness, the appearance of muscle in smaller arteries than normal, and reduction in the number of peripheral arteries. By autoradiographic techniques, after injection of 3H-thymidine into rats fed Crotalaria for 3, 7, 14, 21, 28, or 35 days, the contribution of hyperplasia to these changes has been assessed at two levels of the pulmonary artery--the hilum and the periphery. In the hilar pulmonary artery, a biphasic increase in labeling index (LI) is seen in each cell type. After 3 days of feeding, the medial smooth muscle cells show a slight but significant increase (1.5 times the control value), and, after 7 days, so do the adventitial fibroblasts (3 x) and the endothelial cells (EC) (2 x). After 14 days LI for all three cell types is again at control values, but after 21 days (wall thickness is no increased) each cell type shows at least a fivefold increase; by 35 days all are again near control levels. In the intra-acinar region, by 14 days, "newly" muscularized arteries are identified and increase in number and proportion up to 35 days; 3H-thymidine uptake is not evident in this cell type until 35 days have passed. The ECs of these arteries, however, show a striking increase in LI after 14 days as do those of the alveolar capillaries. The ECs of the intra-acinar veins show a biphasic response being increased after 7, 28, and 35 days. The present study has shown that Crotalaria ingestion induces hyperplasia and hypertrophy of pulmonary arterial cells at pre- and intra-acinar levels. The early increase in LI probably represents a response to the original cell injury, the later changes, a response to continuing damage or, in part, adaptation to the pulmonary hypertension now present.


Chinese Preschool Children's Socioemotional Development: The Effects of Maternal and Paternal Psychological Control.
The present study examined the relative prediction and joint effects of maternal and paternal psychological control on children's socioemotional development. A total of 325 preschool children between the ages of 34 and 57 months (M = 4 years 2 months) and their parents participated in the study. Fathers and mothers, respectively, reported their levels of psychological control and mothers evaluated the socioemotional development of children using two indicators (i.e., behavioral problems and prosocial behaviors). The results indicated that the relative predictive effects of maternal and paternal psychological control on children's socioemotional development differed. Specifically, maternal psychological control was a significant predictor of children's behavioral problems and prosocial behaviors, whereas the levels of paternal psychological control were unrelated to children's socioemotional development. With regard to the combined effects of maternal and paternal psychological control, the results of ANOVAs and simple slope analysis both indicated that children would be at risk of behavioral problems as long as they had one highly psychologically controlling parent. High levels of paternal psychological control were associated with increased behavioral problems of children only when maternal psychological control was low. However, the association between maternal psychological control and children's behavioral behaviors was significant, despite paternal psychological control.


IFNAR2 Is Required for Anti-influenza Immunity and Alters Susceptibility to Post-influenza Bacterial Superinfections.
Influenza virus infections particularly when followed by bacterial superinfections (BSI) result in significant morbidities and mortalities especially during influenza pandemics. Type I interferons (IFNs) regulate both anti-influenza immunity and host susceptibility to subsequent BSIs. These type I IFNs consisting of, among others, 14 IFN-α's and a single IFN-β, are recognized by and signal through the heterodimeric type I IFN receptor (IFNAR) comprised of IFNAR1 and IFNAR2. However, the individual receptor subunits can bind IFN-β or IFN-α's independently of each other and induce distinct signaling. The role of type I IFN signaling in regulating host susceptibility to both viral infections and BSI has been only examined with respect to IFNAR1 deficiency. Here, we demonstrate that despite some redundancies, IFNAR1 and IFNAR2 have distinct roles in regulating both anti-influenza A virus (IAV) immunity and in shaping host susceptibility to subsequent BSI caused by S. aureus. We found IFNAR2 to be critical for anti-viral immunity. In contrast to Ifnar1-/- mice, IAV-infected Ifnar2-/- mice displayed both increased and accelerated morbidity and mortality compared to WT mice. Furthermore, unlike IFNAR1, IFNAR2 was sufficient to generate protection from lethal IAV infection when stimulated with IFN-β. With regards to BSI, unlike what we found previously in Ifnar1-/- mice, Ifnar2-/- mice were not susceptible to BSI induced on day 3 post-IAV, even though absence of IFNAR2 resulted in increased viral burden and an increased inflammatory environment. The Ifnar2-/- mice similar to what we previously found in Ifnar1-/- mice were less susceptible than WT mice to BSI induced on day 7 post-IAV, indicating that signaling through a complete receptor increases BSI susceptibility late during clinical IAV infection. Thus, our results support a role for IFNAR2 in induction of anti-IAV immune responses that are involved in altering host susceptibility to BSI and are essential for decreasing the morbidity and mortality associated with IAV infection. These results begin to elucidate some of the mechanisms involved in how the individual IFNAR subunits shape the anti-viral immune response. Moreover, our results highlight the importance of examining the contributions of entire receptors, as individual subunits can induce distinct outcomes as shown here.


Nanoscale insight into silk-like protein self-assembly: effect of design and number of repeat units.
By means of replica exchange molecular dynamics simulations we investigate how the length of a silk-like, alternating diblock oligopeptide influences its secondary and quaternary structure. We carry out simulations for two protein sizes consisting of three and five blocks, and study the stability of a single protein, a dimer, a trimer and a tetramer. Initial configurations of our simulations are β-roll and β-sheet structures. We find that for the triblock the secondary and quaternary structures upto and including the tetramer are unstable: the proteins melt into random coil structures and the aggregates disassemble either completely or partially. We attribute this to the competition between conformational entropy of the proteins and the formation of hydrogen bonds and hydrophobic interactions between proteins. This is confirmed by our simulations on the pentablock proteins, where we find that, as the number of monomers in the aggregate increases, individual monomers form more hydrogen bonds whereas their solvent accessible surface area decreases. For the pentablock β-sheet protein, the monomer and the dimer melt as well, although for the β-roll protein only the monomer melts. For both trimers and tetramers remain stable. Apparently, for these the entropy loss of forming β-rolls and β-sheets is compensated for in the free-energy gain due to the hydrogen-bonding and hydrophobic interactions. We also find that the middle monomers in the trimers and tetramers are conformationally much more stable than the ones on the top and the bottom. Interestingly, the latter are more stable on the tetramer than on the trimer, suggesting that as the number of monomers increases protein-protein interactions cooperatively stabilize the assembly. According to our simulations, the β-roll and β-sheet aggregates must be approximately equally stable.


Choosing wisely: the Canadian Rheumatology Association's list of 5 items physicians and patients should question.
To develop a list of 5 tests or treatments used in rheumatology that have evidence indicating that they may be unnecessary and thus should be reevaluated by rheumatology healthcare providers and patients. Using the Delphi method, a committee of 16 rheumatologists from across Canada and an allied health professional generated a list of tests, procedures, or treatments in rheumatology that may be unnecessary, nonspecific, or insensitive. Items with high content agreement and perceived relevance advanced to a survey of Canadian Rheumatology Association (CRA) members. CRA members ranked these top items based on content agreement, effect, and item ranking. A methodology subcommittee discussed the items in light of their relevance to rheumatology, potential effect on patients, and the member survey results. Five candidate items selected were then subjected to a literature review. A group of patient collaborators with rheumatic diseases also reviewed these items. Sixty-four unique items were proposed and after 3 Delphi rounds, this list was narrowed down to 13 items. In the member-wide survey, 172 rheumatologists responded (36% of those contacted). The respondent characteristics were similar to the membership at large in terms of sex and geographical distribution. Five topics (antinuclear antibodies testing, HLA-B27 testing, bone density testing, bone scans, and bisphosphonate use) with high ratings on agreement and effect were chosen for literature review. The list of 5 items has identified starting points to promote discussion about practices that should be questioned to assist rheumatology healthcare providers in delivering high-quality care.


[Dilated cardiomyopathy and panuveitis as presenting symptoms of Lyme disease. General review of one case].
The clinical expression of Lyme disease is highly variable. If a patient presents clinical findings consistent with a systemic Lyme borreliosis, this disease must be considered in an endemic area because of its favorable outcome with adequate treatment. The authors report and discuss the case of a patient with an unusual history of dilated cardiomyopathy and supraventricular fibrillation followed by bilateral panuveitis. Enzyme-linked immunosorbent assay and Western blot were positive for Borrelia burgdorferi antigens. The diagnosis of Lyme disease was made after other infectious, inflammatory and autoimmune disorders were excluded by clinical, instrumental and biological investigations. The treatment by ceftriaxone and amoxicillin resolved the ophthalmologic manifestations and improved the cardiac condition. This report underlines the possibility of an unusual presentation of Lyme disease. Ophthalmologic and cardiac involvement should be known by clinicians.


Microbiological analysis of seed sprouts in Norway.
As part of larger survey of microbial contamination of fruits and vegetables in Norway, four different sprouted seed products were analysed for bacterial and parasitic contaminants (n = 300 for bacterial analyses and n = from 17 to 171 for parasite analyses, depending on parasite). Escherichia coli O157, Salmonella, Listeria monocytogenes, Cyclospora oocysts, Ascaris eggs and other helminth parasites were not detected in any of the sprout samples. Thermotolerant coliform bacteria (TCB) were isolated from approximately 25% of the sprout samples, with the highest percentage of TCB positive samples in alfalfa sprouts. Most TCB were Enterobacter spp. and Klebsiella. E. coli was isolated from 8 of 62 TCB positive mung bean sprout samples. Cryptosporidium oocysts were detected in 8% of the sprout samples and Giardia cysts were detected in 2% of the samples. All the Cryptosporidium positive samples, and most of the Giardia positive samples, were mung bean sprouts. Parasite concentrations in positive samples were low (between 1 and 3 oocysts/cysts per 50 g sprouts). Sprout irrigation water was also analysed for microbial contaminants. E. coli O157 and L. monocytogenes were not detected. TCB were isolated from approximately 40% of the water samples. Salmonella reading was isolated from three samples of spent irrigation water on 3 consecutive days. Cryptosporidium and Giardia were also isolated from spent irrigation water. Additionally, eight samples of unsprouted mung bean seed were analysed for Cryptosporidium oocysts and Giardia cysts. One or both of these parasites were detected in six of the unsprouted seed samples at concentrations of between 1 and 5 oocysts/cysts per 100 g unsprouted seed. Whilst our results support spent irrigation water as the most suitable matrix for testing for bacteria, unsprouted seed is considered the more useful matrix for analysing for parasite contaminants.


Time-resolved serial crystallography to track the dynamics of carbon monoxide in the active site of cytochrome c oxidase.
Cytochrome c oxidase (CcO) is part of the respiratory chain and contributes to the electrochemical membrane gradient in mitochondria as well as in many bacteria, as it uses the energy released in the reduction of oxygen to pump protons across an energy-transducing biological membrane. Here, we use time-resolved serial femtosecond crystallography to study the structural response of the active site upon flash photolysis of carbon monoxide (CO) from the reduced heme a3 of ba3-type CcO. In contrast with the aa3-type enzyme, our data show how CO is stabilized on CuB through interactions with a transiently ordered water molecule. These results offer a structural explanation for the extended lifetime of the CuB-CO complex in ba3-type CcO and, by extension, the extremely high oxygen affinity of the enzyme.


Neural correlates of binding features within- or cross-dimensions in visual conjunction search: an fMRI study.
The fMRI technique was used to investigate the functional neuroanatomy of binding features within- or cross-dimension during visual conjunction search. Participants were asked to perform feature search (FS; e.g., search for a vertical bar among tilted bars), within-dimension search (WS; e.g., search for an upright T among non-target oriented Ts and Ls), cross-dimension search (CS; e.g., search for an orange vertical bar among blue vertical bars and orange tilted bars), and complex search combining within- and cross-dimension features (WCS; e.g., search for an orange upright T among orange leftward Ts and blue Ls). Reaction times (RTs) taken to decide whether a target was present or absent were faster in the FS than in the WS, CS, and WCS conditions, but did not differ between the latter three conditions. Neuroimaging results revealed a set of fronto-parietal regions, including frontal eye field and intraparietal sulcus, to be consistently activated in conjunction search (WS, CS, and WCS) relative to feature search, suggesting that these regions play a more prominent role in matching visual input against the target template in conjunction search. Furthermore, left occipito-temporal cortex was more activated in within-dimension conjunction search, and bilateral intraparietal sulci were more activated in cross-dimension conjunction search. This suggests that features from the same dimension are 'bound' at a higher stage of the ventral pathway by conjoining the inputs from lower-level neurons, whereas neurons along the intraparietal sulcus appear to be necessary for discerning the presence of cross-dimensional conjunctions.


Roles of nonclinical and clinical data in prediction of 30-day rehospitalization or death among heart failure patients.
Selecting heart failure (HF) patients for intensive management to reduce readmissions requires effective targeting. However, available prediction scores are only modestly effective. We sought to develop a prediction score for 30-day all-cause rehospitalization or death in HF with the use of nonclinical and clinical data. This statewide data linkage included all patients who survived their 1st HF admission (with either reduced or preserved ejection fraction) to a Tasmanian public hospital during 2009-2012. Nonclinical data (n = 1,537; 49.5% men, median age 80 y) included administrative, socioeconomic, and geomapping data. Clinical data before discharge were available from 977 patients. Prediction models were developed and internally and externally validated. Within 30 days of discharge, 390 patients (25.4%) died or were rehospitalized. The nonclinical model (length of hospital stay, age, living alone, discharge during winter, remoteness index, comorbidities, and sex) had fair discrimination (C-statistic 0.66 [95% confidence interval (CI) 0.63-0.69]). Clinical data (blood urea nitrogen, New York Heart Association functional class, albumin, heart rate, respiratory rate, diuretic use, angiotensin-converting enzyme inhibitor use, arrhythmia, and troponin) provided better discrimination (C-statistic 0.72 [95% CI 0.68-0.76]). Combining both data sources best predicted 30-day rehospitalization or death (C-statistic 0.76 [95% CI 0.72-0.80]). Clinical data are stronger predictors than nonclinical data, but combining both best predicts 30-day rehospitalization or death among HF patients.


Discovery and development of pyrethroid insecticides.
Pyrethroid insecticides contain natural pyrethrins extracted from pyrethrum flowers, and their synthetic derivatives, pyrethroids. The present article provides an overview of the structure of natural pyrethrins, and the discovery and development of pyrethroids with an emphasis on the background of selected compounds. The stereochemical relationships among pyrethroid secondary alcohols, and toxicologic and environmental effects of pyrethroids are also discussed. Finally, the pyrethroid resistance of mosquitoes and future aspects of pyrethroids are addressed.


Are women with severely symptomatic brugada syndrome different from men?
Women with Brugada Syndrome. Spontaneous type-1 ECG has been recognized as a risk factor for sudden cardiac death (SCD) in Brugada syndrome (BrS), but studied populations predominantly consisted of men. We sought to investigate whether a spontaneous type-1 ECG pattern was also associated in women with severely symptomatic BrS. Other known risk factors were also examined for gender specificity. Patients with severely symptomatic BrS, defined as resuscitated SCD and/or appropriate implantable cardioverter-defibrillator (ICD) shock, were included from 11 European centers. Clinical data, investigation of family history, 12-lead ECG, and results of electrophysiological study (EPS) were collected. The average follow-up was 4 +/- 3 years. Fifty-eight patients fulfilled the inclusion criteria (mean age 47 +/- 11 years, 8 women). Thirty-six men (72%) but only two women (25%) had a spontaneous type-1 ECG at baseline (P = 0.02). Maximal ST elevation before or after drug challenge was 3.7 +/- 1.3 mm in men versus 2.4 +/- 0.7 mm in women (P = 0.007). The proportion of patients with a family history of SCD or an SCN5A mutation was not significantly different between both groups. Of those patients with high-risk BrS who underwent EPS, 76%(12/25) of men and 50%(2/4) of women had a positive study. In contrast to men, most women with BrS and resuscitated SCD or appropriate ICD shock do not have a spontaneous type-1 ECG pattern. In addition, the degree of ST elevation is less pronounced in women than men. While women represent a lower-risk group overall, risk factors established from a predominantly male population may not be helpful in identifying high-risk females.


Selective screening for gestational diabetes mellitus. Toronto Trihospital Gestational Diabetes Project Investigators.
The usual approach to detecting gestational diabetes mellitus is to screen all pregnant women by measuring their plasma glucose after a 50-g oral glucose load at 24 to 28 weeks' gestation. Women are referred for an oral glucose-tolerance test if the plasma glucose concentration one hour later is > or = 140 mg per deciliter (7.8 mmol per liter). We hypothesized that the efficiency of screening could be enhanced by considering women's risks of gestational diabetes on the basis of their clinical characteristics. We studied 3131 pregnant women who underwent both the screening and the diagnostic tests. We randomly selected data on half the women and used them to derive new screening strategies. We categorized each woman's risk of gestational diabetes mellitus on the basis of her age, body-mass index before pregnancy, and race. We developed strategies that entailed no screening for low-risk women, usual care for intermediate-risk women, and universal screening with lower thresholds -- plasma glucose values of 130 mg per deciliter (7.2 mmol per liter) or 128 mg per deciliter (7.1 mmol per liter) -- for high-risk women. The strategies were validated with data on the other half of the women. The new strategies allowed a 34.6 percent reduction in the number of screening tests performed (95 percent confidence interval, 32.3 to 37.0) and detected 81.2 to 82.6 percent of the women with gestational diabetes as compared with the 78.3 percent detected through usual care. The percentage of false positive screening tests was significantly reduced, from 17.9 percent with usual care to 16.0 per cent (P=0.02) or 15.4 percent (P<0.001) with the new strategies, depending on the threshold values for high-risk women. Consideration of women's clinical characteristics allows efficient selective screening for gestational diabetes.


Escherichia coli K-12 mutants deficient in uracil-DNA glycosylase.
A new assay specific for uracil-DNA glycosylase is described, Escherichia coli mutants partially and totally deficient in uracil-DNA glycosylase activity have been isolated by using this assay in mass-screening procedures. These have been designated ung mutants. The ung gene maps between tyrA and nadB on the E. coli chromosome. T4 phage containing uracil in their DNA grow on the most glycosylase-deficient hosts but are unable to grow on wild-type bacteria. This provides a simple spot test for the ung genotype. The ung mutants show slightly higher rates of spontaneous mutation to antibiotic resistance. Taken together, these results suggest a central role for uracil-DNA glycosylase in the initiation of an excision repair pathway for the exclusion of uracil from DNA.


[Group behavior of mice in population cages].
A model of a population cage has been proposed, intended for a quantitative and qualitative estimation of the group behaviour of mice, for determining the type of their interrelations and identifying the dominant in the group. In experiments on 120 mice the fundamental types of interaction between mice (peaceful or conflicting) have been singled out in the apparatus, as well as the behaviour phases of each type of interaction. It has been shown that the type and phases of interaction in mice depend on the presence of a dominant in the group, the sex and age composition of the group, the degree of adaptation to one another, and the previous experience (being kept in isolation or in a group). The final effect of group interaction is determined not by a simple sum of individual characteristics of the animals, but is subordinated to the rules of zoosocial behaviour, i. e. the formation of stage by stage dominant- subordinate relationships.


In vitro stimulation of presensitized mouse spleen cells with poliovirus type 1, Mahoney, and enhancement of poliovirus-specific hybridomas.
In vivo immunization of BALB/c mice with poliovirus type 1, strain Mahoney, or with its purified polypeptides resulted in 0.2 to 0.5 antigen-specific hybridoma microcultures per 10(6) spleen cells. Stimulation of spleen cells from mice immunized with poliovirus or with its polypeptides in vitro with poliovirus 6 days prior to fusion with the myeloma cells led to a six- to 20-fold increase in the number of positive microcultures, i.e. after stimulation the yield of poliovirus-specific hybridomas was up to 3.8 antigen-specific microcultures per 10(6) spleen cells. The in vitro stimulation of spleen cells primed in vivo was demonstrated by the detection of poliovirus-specific antibody-producing cells 6 days after in vitro cultivation in the presence of poliovirus as antigen. Only spleen cells stimulated under these conditions in vitro gave rise to specific antibody-producing cells and yielded antigen-specific hybridomas after somatic hybridization.


The development of antibodies to penicilin in rabbits.
A method for the production of antibodies specifically directed against penicillin is described. The inability of this antibody to significantly reduce the antibiotic activity of penicillin is noted. Evidence to show the variability of specificities of various sera, some directed for the most part against the side chain, others against the nucleus is presented. Studies on serum fractions separated electrophorectically indicate that the antibody migrates in the fast gamma-globulin and beta-globulin fractions and requires dextran or albumin to effect agglutination. The inability of penicilloic acid, an hydrolysis product of penicillin to provoke antibody formation despite its ability to inhibit the antibody is shown and the implications of this observations are discussed.


Structure of nuclear pre-mRNA. VIII. Isolation and characterization of complementary sequences.
After the annealing of pre-mRNA at high Cot values, up to 20% of the material becomes resistant to the action of ribonuclease. This has been attributed to the presence of self-complementary sequences (scRNA) in the pre-mRNA. The most important properties of scRNA, which was isolated in preparative amounts, have been studied. The approximate dimensions of the complementary segments are 45-50 nucleotides. Hybridization experiments have shown that scRNA is transcribed from repeated segments of the genome. It may be postulated that the rapidly hybridizing pre-mRNA fraction consists mainly of self-complementary sequences.