[Dyspnea. I. Definition and pathophysiological problems].
An analytical review was made on the pathophysiological mechanisms suspected to be involved in dyspnea, accepting that the sensation of shortness of breathing, and the subjective signs (exertion, anxiety and fear) are essential for the diagnosis of this condition. Bearing in mind the anatomical relations in the central nervous system between the structures involved in the control and regulation of the respiration, and these involved in the emotions, a scheme was presented of the pathophysiological mechanisms, which determine the essential signs of the dyspnea. Three neuronal circuits were assumed for the explanation of the manifestations in dyspnea: subjective signs, symptoms of the thoraco-pulmonary and cardio-vascular systems, and these of the voluntary muscular system that accompanied dyspnea.


The place of newer oral anticoagulants in the treatment of patients with non-valvular atrial fibrillation and coronary artery disease.
With the arrival of the newer oral anticoagulants (NOACs), we now have a more convenient means of providing anticoagulation for patients with non-valvular atrial fibrillation (AF). Are there any particular considerations for the many patients who also have coronary artery disease? For many patients there is an obvious need to use the newer agents but for others, such as those who simply refuse to consider using a vitamin K antagonist (VKA) such as warfarin, the decision may be more problematical.


Leisure Boredom, Timing of Sexual Debut, and Co-Occurring Behaviors among South African Adolescents.
Sex during adolescence is normative; however, there are substantial individual differences in the timing and context of sexual debut. Leisure boredom is an underexplored correlate of sexual behavior that is associated with many adolescent health outcomes. We investigated if and how individual differences in leisure boredom may be associated with timing of sexual debut, and whether individuals engage in safe or risky behaviors at debut. Survival analysis, logistic regression, and Poisson regression were applied to eight-wave longitudinal data obtained from 3,088 South African adolescents (baseline Mage = 13.9 years) to examine associations between leisure boredom and cumulative hazard of sexual debut across adolescence, odds of co-occurring sexual behaviors, and incidence rate of co-occurring sexual risk behaviors at debut. Higher levels of leisure boredom were associated with elevated hazard cumulatively across adolescence. Higher levels of leisure boredom were also associated with lower odds of safe sex and higher odds of substance use during sex and transactional sex at sexual debut, but not casual sex or condom non-use at sexual debut. Although odds of singular risk behaviors were lower for girls than for boys, the association between leisure boredom and the number of risk behaviors at sexual debut was stronger for girls than boys. Higher trait leisure boredom was associated with elevated hazard of sexual debut, greater likelihood that risky behaviors accompanied sexual debut, and greater number of co-occurring risky behaviors at sexual debut. Results support leisure boredom as a potential target for preventing sexual risk behavior among South African adolescents.


Sexually Explicit Internet Media Consumption and Sexual Risk Behaviors Among Chinese Male Sexual Minorities: The Moderating Role of Perceived Realism.
Western research has suggested that viewing sexually explicit internet media (SEIM) may result in sexual risk behaviors (SRBs) among gay, bisexual, and other men who have sex with men. This study explored the relationship between SEIM consumption and SRBs and whether perceived realism could moderate this relationship among Chinese male sexual minorities. We assessed SEIM consumption from two aspects: consumption frequency and preference for specific content (condomless and group sex). This study recruited 360 participants from various online channels. Consistent with the sexual script acquisition, activation, application model (3AM), we found that both SEIM consumption frequency and content preference were related to SRBs, and that perceived realism moderated the relationship between SEIM consumption and SRBs. Preference for group sex SEIM was significantly associated with an increased likelihood of casual sex when perceived realism was higher. SEIM use frequency was significantly associated with an increased likelihood of unprotected anal intercourse when perceived realism was higher. These findings may be used to promote HIV prevention, which necessitates joint effort from multiple stakeholders.


Fungal organic acid uptake of mineral-derived K is dependent on distance from carbon hotspot.
Fungal species are foundational members of soil ecosystems with vital contributions that support interspecies resource translocation. The minute details of these biogeochemical processes are poorly investigated. Here, we addressed this knowledge gap by probing fungal growth in a novel mineral-doped soil micromodel platform using spatially-resolved imaging methodologies. We found that fungi uptake K from K-rich minerals using organic acids exuded in a distance-dependent manner from a carbon-rich hotspot. While identification of specific mechanisms within soil remains challenging, our findings demonstrate the significance of reduced complexity platforms such as the mineral-doped micromodel in probing biogeochemical processes. These findings provide visualization into hyphal uptake and transport of mineral-derived nutrients in a resource-limited environment.


[Suicide re-attempt risk evaluation scale: a trial in a critical care center without an exclusive duty psychiatrist].
There is a general consensus that patients who attempt suicide require intervention for psychological as well as physical problems. However, among the institutions accepting patients who attempt suicide, there are few institutions employing a full-time psychiatrist in the emergency department. Therefore, it is difficult to conduct a suicide re-attempt risk evaluation in the presence of a psychiatric specialist. We developed a suicide re-attempt risk evaluation scale to be used by non-professional staff. We asked the medical staff that cared for patients who had attempted suicide to record all the evaluation items in the patient's medical record. We utilized an interview method and highlighted the types of answers that needed special attention. When there were two or more answers that needed special attention, we defined the patient as high risk. The number of days of hospitalization did not change as a result of the use of the suicide re-attempt risk estimate scale; however, the quantity of medical records addressing suicide intention and the rate of psychiatric intervention significantly increased. It is unclear whether this score directly led to the prevention of repeat suicide attempts. However, it is notable that use of the scale has led to a change in the behavior of medical staff by mandating a fixed-format risk evaluation.


Hypothalamic Glioma in a Patient With Sturge-Weber Syndrome.
Sturge-Weber syndrome (SWS) is a neurocutaneous disorder characterized by vascular malformations involving brain, skin, and occasionally eyes. There is no recognized tumor predisposition in patients with SWS as there is with some other phakomatoses. We present a patient with SWS who developed a low-grade glioma (LGG). We hypothesize that there could be an association between SWS and LGG formation, noting that GNAQ mutations have been implicated in the underlying biology of both SWS and a subset of pediatric LGG. It is suggested that SWS may be a cancer predisposition syndrome.


Red cells from ferrochelatase-deficient erythropoietic protoporphyria patients are resistant to growth of malarial parasites.
Many red cell polymorphisms are a result of selective pressure by the malarial parasite. Here, we add another red cell disease to the panoply of erythrocytic changes that give rise to resistance to malaria. Erythrocytes from individuals with erythropoietic protoporphyria (EPP) have low levels of the final enzyme in the heme biosynthetic pathway, ferrochelatase. Cells from these patients are resistant to the growth of Plasmodium falciparum malarial parasites. This phenomenon is due to the absence of ferrochelatase and not an accumulation of substrate, as demonstrated by the normal growth of P falciparum parasites in the EPP phenocopy, X-linked dominant protoporphyria, which has elevated substrate, and normal ferrochelatase levels. This observation was replicated in a mouse strain with a hypomorphic mutation in the murine ferrochelatase gene. The parasite enzyme is not essential for parasite growth as Plasmodium berghei parasites carrying a complete deletion of the ferrochelatase gene grow normally in erythrocytes, which confirms previous studies. That ferrochelatase is essential to parasite growth was confirmed by showing that inhibition of ferrochelatase using the specific competitive inhibitor, N-methylprotoporphyrin, produced a potent growth inhibition effect against cultures of P falciparum. This raises the possibility of targeting human ferrochelatase in a host-directed antimalarial strategy.


Self-Boosted Tunnel Field-Effect Transistor Using Nitride Charge Trapping Layer for Low Supply Voltage Operation.
Tunneling field-effect transistors (TFETs) have been studied as a candidate for low-power device due to the remarkable subthreshold characteristics. However, digital circuits composed of TFET have significantly large propagation delay compared with the conventional MOSFET circuits because of small current drivability and large gate-to-drain capacitance. In this work, the electrical characteristics of the self-boosted TFETs with nitride charge trapping layer have been studied using TCAD simulations. Trapped charges in the nitride layer improve subthreshold characteristics and on-current (I(ON)) of both nTFET and pTFET during gate bias sweep. In addition, the benefits of the self-boosted TFET devices to low supply voltage system application are investigated. Energy consumption and propagation delay of both conventional and self-boosted TFET inverters are compared by the mixed-mode circuit simulation study. Energy consumption is almost same but the propagation delay of the self-boosted TFET inverter is reduced especially for ultra-low voltage operation where system delay is increased dramatically.


Association between bowel habits and quality of bowel preparation for colonoscopy.
The effectiveness of colonoscopy is highly dependent on the quality of bowel preparation. Although many studies have previously evaluated the role of cleansing methods and dosing regimens, few have examined the association between bowel habits and subsequent bowel preparation. Here, we aimed to evaluate the impact of bowel habits on the quality of bowel preparation.A total of 404 patients who underwent a total colonoscopy and completed a personal bowel habit questionnaire at Korea University Hospital between December 2012 and December 2013 were enrolled. The usual stool form of patients was classified into 7 categories according to the Bristol Stool Scale (BSS). The quality of bowel preparation was determined during colonoscopy according to the Ottawa Bowel Preparation Scale (OBPS). Segment scores of ≥3 or total OBPS scores of >7 were defined as poor bowel preparation.Poor bowel preparation was reported in 9.4% of observed colonoscopies. The odds ratio (OR) of poor bowel preparation being associated with infrequent bowel movements (<3/week) was 5.00 (95% confidence interval [CI], 1.91-13.1, P = .001). BSS types 1 and 2 tended to have an association with poor bowel preparation, but the association was statistically insignificant (OR: 2.38; 95% CI, 0.90-6.33, P = .082). After adjusting for age, sex, drinking, presence of diabetes mellitus, and bowel preparation regimen, infrequent bowel movement (<3/week) was still significantly associated with poor bowel preparation. When subdividing by colonic segment, it was significantly associated with poor bowel preparation in all segments.Infrequent bowel movement (<3/week) was significantly associated with poor bowel preparation.


Maternal and gestational factors in cryptorchidism.
Previous epidemiological studies of cryptorchidism have led to the hypothesis that the risk of undescended testis is associated with excess oestrogen exposure during pregnancy. A case-control study was undertaken to test this hypothesis, comparing mothers of affected boys (244) and normal male births (488) born within six months of a case selected randomly from the British Columbia population. Information was collected on the mother's reproductive history, family history, and past medical history, and events surrounding all pregnancies ending in a birth. The results were analysed using both the population-based sample of male births and the male sibs of cases as control groups. Neither exogenous oestrogen exposure, nor any of the pregnancy-related variables hypothesized to be indirect indicators of endogenous oestrogen exposure, including bleeding and nausea and/or vomiting, were found to be significantly associated with risk of undescended testes in either comparison. More mothers with later index births reported menstrual irregularity greater than half the time, and smoking, thought to have a protective effect, was more prevalent among case mothers than control mothers. No other variables were significantly different between case and control mothers. The results of this study do not support the hypothesis that elevated exogenous or endogenous oestrogen exposure during pregnancy increases the risk of undescended testis in male children.


Evolution of the hepatitis E virus polyproline region: order from disorder.
The hepatitis E virus (HEV) polyproline region (PPR) is an intrinsically unstructured region (IDR). This relaxed structure allows IDRs, which are implicated in the regulation of transcription and translation, to bind multiple ligands. Originally the nucleotide variability seen in the HEV PPR was assumed to be due to high rates of insertion and deletion. This study shows that the mutation rate is about the same in the PPR as in the rest of the nonstructural polyprotein. The difference between the PPR and the rest of the polyprotein is due to the higher tolerance of the PPR for substitutions at the first and second codon positions. With this higher promiscuity there is a shift in nucleotide occupation of these codons leading to translation of more cytosine residues: a shift that leads to more proline, alanine, serine, and threonine being encoded rather than histidine, phenylalanine, tryptophan, and tyrosine. This pattern of amino acid usage is typical of proline-rich IDRs. Increased usage of cytosine also leads to >22% of all amino acids in the PPR being prolines. Alignments of PPR sequences from HEV strains representing all genotypes indicate that all zoonotic isolates share an ancestor, and the carboxyl half of the PPR is more tolerant of mutations than the amino half. The evolution of HEV PPR, in contrast with that of the rest of the nonstructural polyprotein, is molded by pressures that lead toward increased proline usage with a corresponding decrease in the usage of aromatic amino acids, favoring formation of IDR structures.


[Immunofluorescent localization of associated antigen of anti-human nasopharyngeal carcinoma cell monoclonal antibody].
The immunofluorescent localization of associated antigen of anti-human nasopharyngeal carcinoma (NPC) cell monoclonal antibody (McAb) was performed in different tissues (NPC, non-NPC tumor, chronic inflammation of nasopharyngeal mucosa, adult normal tissue and embryo tissue) with 5 control experiments. McAb (CN-1, Cs-C1) was produced by Department of Microbiology of our college and all the specimens were confirmed by pathology. The results revealed that Cs-C1 antigen was mainly found on the surface of NPC cells with a positive rate of 80.3%. Majority of the positive cells showed yellow-greenish linear fluorescence surrounding the cell membrane while some cells manifested granular fluorescence. In addition, Cs-C1 antigen was also found in a few embryo tissues, epithelial cells of the normal gastric mucosa and cells of the gastric cancer. It is suggested that Cs-C1 antigen be a kind of molecular structure, being gradually produced or increased in quantity during carcinogenesis, and belong to the tumor associated antigen. Cs-C1 antigen could be important in the study of carcinogenic mechanism of NPC cells and valuable to clinical immunodiagnosis.


Albumin to ascites: demonstration of a direct pathway bypassing the systemic circulation.
The transport of plasma albumin and newly made albumin into ascitic fluid was studied in eight patients with cirrhosis and ascites. The thoracic duct was cannulated in two patients and lymph collected over a period of 2 hr. Simultaneously albumin-(131)I and carbonate-(14)C were injected intravenously. The albumin-(131)I measured the transfer of plasma albumin into ascites and into thoracic duct lymph. The carbonate-(14)C, by labeling newly formed albumin, permitted the estimation of the transfer of newly formed albumin into plasma, ascites, and lymph. If the newly synthesized albumin entering ascites and thoracic duct lymph is delivered initially into the plasma, then the ratios of the albumin-(14)C and -(131)I in ascites and lymph compared with the content of albumin-(14)C and -(131)I in plasma would be identical. However, if some newly formed albumin is delivered directly into ascites or lymph, the ratio for albumin-(14)C would be higher than that for albumin-(131)I in lymph or ascites. The ratios of both labeled albumins found in ascites or lymph are expressed as per cent of the total plasma pool. In the eight patients studied 4.2-11.7% of the albumin-(14)C in plasma was found in ascites in 2 hr whereas only 0.4-2.2% of plasma albumin-(131)I entered in this same period. In the two patients studied during thoracic duct lymph drainage 6.1 and 13.5% of newly made albumin-(14)C appeared in lymph in 2 hr whereas only 2.8 and 3.8% of plasma albumin-(131)I was found in the lymph. In cirrhosis with ascites some newly formed albumin entered ascites and thoracic duct lymph by a direct pathway from the liver bypassing the systemic circulation.


Haemostatic agent etamsylate in vitro and in vivo antagonizes anti-coagulant activity of heparin.
Etamsylate is indicated for several anti-hemorrhagic indications in human and veterinary medicine. However, etamsylate has been shown to be effective only in specific hemorrhagic situations. Furthermore, mechanism of action of etamsylate is not known but recent research has shown its ability to inhibit heparin binding to several growth factors. We have evaluated the ability of etamsylate to interfere with the activities of heparin. Effects of etamsylate on vasodilatory activity of heparin were evaluated in rat aortic segments. Influence of etamsylate on anticoagulant activity of heparin was evaluated in vitro by determining prothrombin (PT) time and activated partial thromboplastin time (aPTT) in dog blood and in vivo by determining the interference of systemic and topical etamsylate on heparin-induced extension in bleeding time (BT) in rats. Despite failing to inhibit heparin-induced vasodilation of rat aorta, etamsylate significantly reduced the increase in aPTT caused by heparin (+30.4 ± 6.7% vs. +15.0 ± 2.8% for etamsylate at 100 µM, P < 0.05). Etamsylate also antagonized the anticoagulant effects driven by heparin in vivo since prevented the heparin-induced increase in BT when systemically (i.p.) administered (+94.6 ± 7.5% vs. +57.9 ± 9.2% at 10 mg/kg, P < 0.05, vs. +22.2 ± 16.8% at 30 mg/kg, P < 0.01). Additionally, topically applied etamsylate (125 mg/ml) significantly reduced heparin-induced BT increase (+102.5 ± 3.2% vs. +54.0 ± 5.8%, P < 0.01). These evidences show a pharmacological interference by etamsylate on heparin activities antagonizing pro-hemorrhagic effects of heparin in vitro and in vivo without inhibiting its vasodilatory properties. This ability could help to explain pharmacological effects of etamsylate and proposes its role for reversing pro-hemorrhagic states.


Optical Measurement of Stem Xylem Vulnerability.
The vulnerability of plant water transport tissues to a loss of function by cavitation during water stress is a key indicator of the survival capabilities of plant species during drought. Quantifying this important metric has been greatly advanced by noninvasive techniques that allow embolisms to be viewed directly in the vascular system. Here, we present a new method for evaluating the spatial and temporal propagation of embolizing bubbles in the stem xylem during imposed water stress. We demonstrate how the optical method, used previously in leaves, can be adapted to measure the xylem vulnerability of stems. Validation of the technique is carried out by measuring the xylem vulnerability of 13 conifers and two short-vesseled angiosperms and comparing the results with measurements made using the cavitron centrifuge method. Very close agreement between the two methods confirms the reliability of the new optical technique and opens the way to simple, efficient, and reliable assessment of stem vulnerability using standard flatbed scanners, cameras, or microscopes.


Anisotropic Carbon Nanotube Structures with High Aspect Ratio Nanopores for Li-Ion Battery Anodes.
Technological advances in membrane technology, catalysis, and electrochemical energy storage require the fabrication of controlled pore structures at ever smaller length scales. It is therefore important to develop processes allowing for the fabrication of materials with controlled submicron porous structures. We propose a combination of colloidal lithography and chemical vapor deposition of carbon nanotubes to create continuous straight pores with diameters down to 100 nm in structures with thicknesses of more than 300 μm. These structures offer unique features, including continuous and parallel pores with aspect ratios in excess of 3000, a low pore tortuosity, good electrical conductivity, and electrochemical stability. We demonstrate that these structures can be used in Li-ion batteries by coating the carbon nanotubes with Si as an active anode material.


Interaction of met-enkephalin with human granulocytes.
Met-enkephalin has been found to have an effect on cell shape and motility of human polymorphonuclear leukocytes (PMNs). Specific binding of tritium-labeled Met-enkephalin to the cells could not be demonstrated. There was a rapid proteolytic degradation of the peptide in the medium, followed by uptake of the labeled tyrosine. The peptidase recognizes the N-terminal sequence of various endogenous opioid peptides. The protease inhibitors bestatin and bacitracin had but little effect on the degradation of Met-Enk.


Hormone replacement therapy and prognosis in ovarian cancer patients.
Estrogen exposure has at least a moderate effect on the risk for ovarian cancer, and antiestrogen therapy may be helpful in treating the disease. It is known from breast cancer that previous hormone replacement therapy (HRT) may influence the molecular profile and prognostic behavior of these tumors. The aim of this study was therefore to investigate the influence of previous HRT on the prognosis in a cohort of patients with invasive epithelial ovarian cancer. Among 547 patients who were treated for ovarian malignancies at a single institution from 1995 to 2008, a total of 244 postmenopausal patients with epithelial cancer and under the age of 75 were identified for whom information about HRT before the onset of the disease was available. HRT was correlated with tumor and patient characteristics. Analyses of overall survival and progression-free survival were carried out using Cox proportional hazards models. Age, tumor stage, and resection status correlated significantly with HRT in the univariate analysis. Patients with previous HRT were more likely to have a lower stage, to be younger, and to have optimal debulking. With regard to survival, HRT had a positive effect on overall survival, specifically in the subgroup of patients with optimal debulking. No correlation was seen in relation to progression-free survival. Sex hormone exposure through HRT may influence the behavior of ovarian cancers after the onset of the disease. This study supports the hypothesis that ovarian cancer is a hormonally influenced tumor.


Results of screw/wire/polymethylmethacrylate composite fixation for acetabular fracture repair in 14 dogs.
To assess the clinical results in dogs with acetabular fractures stabilized using a screw-wire-polymethylmethacrylate (SWP) composite fixation. A retrospective study of client-owned dogs with acetabular fractures. Fourteen dogs ranging in age from 4 to 95 months (mean, 34 +/- 25 months; median, 25 months) and body weight from 8 to 39 kg (mean, 25 +/- 6 kg; median, 27 kg). Medical records and radiographs were retrospectively evaluated to determine location of the fracture, presence of preexisting degenerative joint disease, accuracy of fracture reduction and complications associated with surgery. Long-term results were evaluated by subjective assessment of lameness, elicitation of pain and/or crepitus on manipulation of the coxofemoral joint, measurements of pelvic limb circumference, coxofemoral joint goniometric measurements, and radiographic evaluation. Fracture reduction was considered anatomic in 13 dogs. At the time of the last follow-up evaluation (mean, 347 +/- 261 days; median, 380 days) 10 dogs were sound on the affected limb, three dogs had a subtle weight-bearing lameness of the affected limb, and the remaining dog had a consistent non-weight-bearing lameness of the affected limb. Mild (n = 10) or moderate (n = 1) degenerative changes of the affected coxofemoral joint attributed to the acetabular fracture and its repair were noted on the follow-up radiographs in 11 dogs. Limb circumference of the affected limb ranged from -8.2% to +10.8% (mean, -0.8 +/- 4.2%; median, -0.7%) of the contralateral limb. The SWP composite fixation consistently maintained anatomic reduction, was associated with few complications, and yielded satisfactory clinical results. The SWP composite fixation technique would seem to be an acceptable means of stabilizing acetabular fractures in dogs.


Antibody vs. HIV in a clash of evolutionary titans.
HIV has evolved many strategies to avoid neutralizing antibody responses, particularly to conserved regions on the external glycoprotein spikes of the virus. Nevertheless, a small number of antibodies have been evolved by the human immune system to recognize conserved parts of the glycoproteins, and therefore, have broadly neutralizing cross-strain activities. These antibodies constitute important tools in the quest to design immunogens that can elicit broadly neutralizing antibodies in humans and hence contribute to an effective HIV vaccine. Crystallographic analyses of the antibodies, in many cases in an antigen-complexed form, have revealed novel and, in some instances, remarkable structural adaptations to attain virus recognition. Antibodies, like HIV, can evolve relatively rapidly through mutation and selection. It seems that the structures of these broadly neutralizing antibodies bear witness to a heroic struggle between two titans of rapid evolution.


Fatal fulminant hepatitis in a patient taking abiraterone acetate: a case report.
Abiraterone acetate is a steroidal inhibitor of cytochrome P450 17A1 indicated in the treatment of metastatic prostate cancer. This report examines the case of a 66-year-old patient diagnosed with prostate adenocarcinoma that had metastasized to the bones and lymph nodes. Treatment with abiraterone acetate and corticosteroid co-administration as well as LH-RH analog hormone therapy was initiated. Four and a half months later, the patient consulted for deterioration of general condition. Biologically, he developed a fulminant hepatitis of which he eventually died. An infectious or metabolic origin was ruled out. Oncological cause by either disease progression or second neoplastic process was eliminated by means of imaging. Hepatic toxicity was imputed to the treatment with abiraterone acetate. This case suggests that fulminant hepatitis on abiraterone acetate may be underestimated, and underscores the importance of regular monitoring of liver tests on this therapy.


Gunshot-caused Facial Injury Combined with Lower Cervical Spine Injury: A Case Report.
A 32-year old male patient was wounded by a pistol. As shown in computed tomography (CT) scanning images, there was comminuted fracture of the left mandible and the bullet was found in the left side behind the sixth cervical vertebra. After the patient was hospitalized, the debridement was done in the emergency room and the operation of open reduction and internal fixation for comminuted fracture of left mandible was performed successfully. Eighteen days later, the patient was taken to surgery for anterior cervical decompression and fusion with autogenous iliac bone grafting for the sixth cervical vertebra. Postoperative follow-up of the patient over two years indicated that the left biceps muscle strength was recovered to level 4. Gunshot wound to the face associated with injury of the low cervical spine has the possibility of survival. It is safe to treat facial wounds early in the patient's treatment course, even if the bullet remains in the cervical vertebral body and there is neurological function damage.


The effect of conventional vitamin D(2) supplementation on serum 25(OH)D concentration is weak among peripubertal Finnish girls: a 3-y prospective study.
To study the effect of vitamin D supplementation and the impact of summer season on serum 25-hydroxyvitamin D (S-25(OH)D) in Finnish 9-15-y-old girls. Three-year follow-up study with vitamin D(2) supplementation using D(2) 10 microg daily from October to January for the first and from October to February for the second winter as well as 20 microg daily from October to March for the third winter. Paavo Nurmi Centre, University of Turku, Turku, Finland. A total of 171 female volunteers aged 9-15 y. Vitamin D and calcium intakes were estimated by a semi-quantitative food frequency questionnaire (FFQ). S-25(OH)D was measured by radioimmunoassay. The median daily dietary intakes of vitamin D and calcium were 3.8 microg (interquartile range (IQR) 2.7-5.0) and 1451 mg (IQR 1196-1812), respectively, over 3 y. The prevalence of severe hypovitaminosis D (S-25(OH)D<20 nmol/l) was 14% and of moderate hypovitaminosis D (20 nmol/l < or = S-25(OH)D < or = 37.5 nmol/l) 75% at baseline in winter. None of the participants had severe hypovitaminosis D in summer. The effect of 10 microg of D(2) daily was insufficient to raise S-25(OH)D from baseline. The daily supplementation of 20 microg of D(2) increased S-25(OH)D significantly in wintertime compared with the non-supplement users (to 45.5 vs 31.8 nmol/l; P<0.001). None of the subjects with vitamin D(2) supplementation approximately 20 microg daily had severe hypovitaminosis D; however, 38% of those participants had moderate hypovitaminosis D at 36 months. Sun exposure in summer raised mean S-25(OH)D to 62.0 nmol/l. Both the daily supplementation of approximately 20 microg of D(2) and summer sunlight exposure had more effect on those who had severe hypovitaminosis than those who had a normal vitamin D status (increase of 24.2 vs 0.9 nmol/l (P<0.001), and 38.8 vs 18.2 nmol/l (P<0.001), respectively). Vitamin D supplementation daily with 20 microg is needed to prevent hypovitaminosis D in peripubertal Finnish girls in winter. Sunlight exposure in summer is more effective than approximately 20 microg of D(2) supplementation daily in winter to raise S-25(OH)D. Both the daily supplementation with 20 microg of D(2) and summertime sunlight exposure had more effect on those who had severe hypovitaminosis D than those who had a normal vitamin D status. Supported by the Yrjö Jahnsson Foundation and the Medical Research Foundation of the Turku University Central Hospital.


Platinum concentration in sentinel lymph nodes after preoperative intra-arterial cisplatin chemotherapy targeting primary tongue cancer.
We conclude that intra-arterially injected cisplatin passed via lymph flow into sentinel nodes (SNs) as the platinum concentration in the SNs was higher than that in the non-sentinel nodes (NSNs). It is possible that preoperative intra-arterial chemotherapy targeting primary cancer also has a therapeutic effect on subclinical metastatic SNs. Intra-arterial chemoradiotherapy has been reported to be effective against not only primary tumors but also nodal metastases. We considered the hypothesis that intra-arterially injected cisplatin passed via lymph flow into regional nodes. This study aimed to investigate intra-arterially injected cisplatin distribution to regional nodes by comparing platinum concentrations in SNs and NSNs. Five patients with T1-2 N0 tongue cancer were treated with preoperative intra-arterial chemotherapy (cisplatin, 100 mg/m(2)) targeting primary cancer. Partial glossectomy together with SN biopsy and elective neck dissection were performed 2 weeks after intra-arterial chemotherapy. Platinum concentrations in the lymph nodes were measured using a Zeeman atomic absorption spectrometer. Thirteen SNs were harvested together with eight NSNs from the areas adjacent to the SNs. Platinum concentrations were then measured, revealing a significant difference in platinum concentration between the SNs and the NSNs (mean ± SD, 0.682 ± 0.246 µg/g vs 0.506 ± 0.274 µg/g; p = 0.049).


Impact of poverty, not seeking medical care, unemployment, inflation, self-reported illness, and health insurance on mortality in Jamaica.
An extensive review of the literature revealed that no study exists that has examined poverty, not seeking medical care, inflation, self-reported illness, and mortality in Jamaica. The current study will bridge the gap by providing an investigation of poverty; not seeking medical care; illness; health insurance coverage; inflation and mortality in Jamaica. Using two decades (1988-2007), the current study used three sets of secondary data published by the (1) Planning Institute of Jamaica and the Statistical Institute of Jamaica (Jamaica Survey of Living Conditions) (2) the Statistical Institute of Jamaica (Demographic Statistics) and (3) the Bank of Jamaica (Economic Report). Scatter diagrams were used to examine correlations between the particular dependent and independent variables. For the current study, a number of hypotheses were tested to provide explanation morality in Jamaica. The average percent of Jamaicans not seeking medical care over the last 2 decades was 41.9%; and the figure has been steadily declining over the last 5 years. In 1990, the most Jamaicans who did not seek medical care were 61.4% and this fell to 52.3% in 1991; 49.1% in 1992 and 48.2% the proceeding year. Based on the percentages, in the early 1990s (1990-1994), the percent of Jamaicans not seeking medical care was close to 50% and in the latter part of the decade, the figure was in the region of 30% and the low as 31.6% in 1999. In 2006, the percent of Jamaicans not seeking medical care despite being ill was 30% and this increased by 4% the following year. Concomitantly, poverty fell by 3.1 times over the 2 decades to 9.9% in 2007, while inflation increased by 1.9 times, self-reported illness was 15.5% in 2007 with mortality averaging 15,776 year of the 2 decades. There is a significant statistical correlation between not seeking medical-care and prevalence of poverty (r = 0.759, p< 0.05). There is a statistical correlation between not seeking medical care and unemployment; but the association is a non-linear one. The relationship between mortality and unemployment was an unsure one, with there being no clear linear or non-linear correlation. The findings revealed that there is a strong direct association between not seeking medical care and inflation rate (r = 0.752). A strong negative statistical correlation was found between mortality and prevalence of poverty (r=0.717). There is a non-linear statistical association between not seeking medical care and illness/injury. Not seeking medical care is not a good indicator of premature mortality; but that this percentage must be excess of 55%. While this study cannot confirm a clear rate of premature mortality, there are some indications that this occurs beyond a certain level of not seeking care for illness.


Topical Quercetin and Resveratrol Protect the Ocular Surface in Experimental Dry Eye Disease.
Purpose: To determine the anti-inflammatory effect of quercetin (QCT), resveratrol (RES), and their combination in a dry eye disease (DED) model. Methods: 0.01% QCT, 0.1% RES, 0.01% QCT + 0.1% RES (QCT + RES) or vehicle were topically applied in a desiccating stress (DS) mice model. CD4+ T cells isolated from DS-exposed mice were transferred to athymic recipient mice. Corneal fluorescein staining, tear production, and tear cytokine levels were evaluated in DS-exposed mice, and conjunctival CD4+ T cell infiltration was evaluated in recipient mice. Results: QCT (p < 0.001) and QCT + RES (p < 0.05) reduced corneal staining in DS-exposed mice. IL-1α tear concentration was reduced by QCT, RES, and QCT + RES (p < 0.05, 0.01 and 0.01, respectively) compared to DS + vehicle mice. CD4+ T cells increased in recipients of DS-exposed mice (p < 0.05) and were lower in recipients of QCT- and RES-treated mice (p < 0.05). Conclusion: The anti-inflammatory effect of QCT, RES, and QCT + RES on DED-experimental model suggests that their topical application could be used for DED treatment.


Evidence for early endosome-like fusion of recently endocytosed synaptic vesicles.
Early endosomes are well-established acceptor compartments of endocytic vesicles in many cell types. Little evidence of their existence or function has been obtained in synapses, and it is generally believed that synaptic vesicles recycle without passing through an endosomal intermediate. We show here that the early endosomal SNARE proteins are enriched in synaptic vesicles. To investigate their function in the synapse, we isolated synaptic nerve terminals (synaptosomes), stimulated them in presence of different fluorescent markers to label the recycling vesicles and used these vesicles in in vitro fusion assays. The recently endocytosed vesicles underwent homotypic fusion. They also fused with endosomes from PC12 and BHK cells. The fusion process was dependent upon NSF activity. Moreover, fusion was dependent upon the early endosomal SNAREs but not upon the SNAREs involved in exocytosis. Our results thus show that at least a fraction of the vesicles endocytosed during synaptic activity are capable of fusing with early endosomes and lend support to an involvement of endosomal intermediates during recycling of synaptic vesicles.


Homeobox gene HB24, a regulator of haematopoiesis, is a candidate for regulating differentiation of the extra-embryonic trophoblast cell lineage.
Expression of the human homeobox gene HB24 in the cytotrophoblast stem cell population in first-trimester human placenta was investigated. HB24 expression was downregulated after cytotrophoblast had differentiated into syncytiotrophoblast. Expression of HB24 was also detected in first-trimester invasive extravillous cytotrophoblast cells. In term placenta, HB24 expression was detected in residual cytotrophoblast cells and in syncytiotrophoblast. Northern analysis revealed two HB24 transcripts in first-trimester placenta of approximately 2.5 kb and 5.3 kb. Ribonuclease protection assays were used to confirm expression of HB24 in first-trimester and term placenta and also to provide evidence that HB24 is downregulated in choriocarcinoma cell lines. In addition to being required for differentiation of the haematopoietic cell lineage, HB24 may be required for differentiation of the extra-embryonic trophoblast cell lineage.


Can the Americans With Disabilities Act Reduce the Death Toll From Police Encounters With Persons With Mental Illness?
A substantial proportion of people shot by police have mental disorders, and many of these killings appear to have been avoidable. One tool to encourage better police training and more cautious behavior is the Americans with Disabilities Act (ADA). However, police groups oppose application of the ADA to arrests, fearing limits on their discretion, and the U.S. Supreme Court appears to favor that view. When the Court declined a recent opportunity to decide the question, it left open a window of opportunity during which the ADA can be leveraged to improve how police officers deal with persons with mental illness.


Light transport in two-layer tissues.
We study theoretically light backscattered by tissues using the radiative transport equation. In particular we consider a two-layered medium in which a finite slab is situated on top of a half space. We solve the one-dimensional problem in which a plane wave is incident normally on the top layer and is the only source of light. The solution to this problem is obtained formally by imposing continuity between the solutions for the upper and lower layers. However, we are interested solely in probing the top layer. Assuming that the optical properties in the lower layer are known, we remove it from the problem yielding a finite slab problem by prescribing an alternate boundary condition. This boundary condition is derived using the theory of Green's functions and is exact. Hence, one needs only to solve the transport equation in a finite slab using this alternate boundary condition. We derive an asymptotic solution for the case when the slab is optically thin. We extend these results to the three-dimensional problem using Fourier transforms. These results are validated by comparisons with numerical solutions for the entire two-layered problem.


Single administration of captopril and combined use with beta-blocker and/or thiazide diuretic in the treatment of essential hypertension.
Thirty-four patients with essential hypertension at WHO stage I or II were divided into three groups. Group I consisted of 22 cases who displayed normal renin activity (NR) or low renin activity (LR) and who received a single administration of captopril. Group II consisted of 6 cases given beta-blockers after administration of captopril. Group III consisted of 6 cases in whom beta-blocker was replaced with thiazide diuretics after administration of captopril alone. Blood pressure decreased significantly by captopril treatment alone in group I of the NR and LR subgroups (except for the diastolic blood pressure [DBP] of the NR subgroup) and fell below the target blood pressure (SBP of 165 mmHg and DBP of 95 mmHg) in 86% of the NR subgroup and 73% of the LR subgroup. Combined treatment with captopril and beta-blocker in Group II did not decrease blood pressure any lower than with captopril alone treatment and achieved the target blood pressure in only 50% of the patients. In group III, combined treatment with captopril and thiazide achieved the target blood pressure in 100% of the patients. Plasma renin activity (PRA) was increased by captopril but reduced by captopril in combination with beta-blocker. However, when beta-blocker was replaced with thiazide, PRA increased. The serum sodium concentration was significantly reduced in the LR subgroup after a single administration of captopril, but there was no other variation.


The novel hydrogen-rich binuclear cobalt complex anion, [(CoH3)2H3](5-).
Hydrogen reacts with a mixture of cobalt bromide and phenylmagnesium bromide to yield a novel cobalt hydride species. EXAFS analysis reveals a binuclear cobalt hydride complex comprising three absorbers, principally a Co-Co centre, and also two types of supporting hydride ligands. Further, DFT, XANES, (1)H NMR spectra and elemental analyses also support the title formulation.


A novel GUCY2D mutation, V933A, causes central areolar choroidal dystrophy.
To identify the genetic cause of central areolar choroidal dystrophy (CACD) in a large Northern Irish family. We previously reported linkage of the locus for CACD in this family to an interval of approximately 5 cM on chromosome 17p13 flanked by polymorphic markers D17S1810 and CHLC GATA7B03. We undertook sequence capture, massively parallel sequencing and computational alignment, base-calling and annotation to identify a causative mutation. Conventional sequencing was used to confirm the Results. Deep sequencing identified a single-base substitution in guanylate cyclase 2D, membrane (retina-specific) gene (GUCY2D). The novel mutation segregated with the disease phenotype and resulted in substitution of valine by alanine at position 933, within the catalytic domain of the protein. It altered a motif that is strongly conserved in a large number of distantly related proteins across several species and was predicted to have a damaging effect on protein activity. Mutations in GUCY2D have previously been associated with dominant cone-rod dystrophies (CORD6) and recessive forms of Leber's congenital amaurosis. This is the first report of a GUCY2D mutation causing CACD and adds to our understanding of genotype-phenotype correlation in this heterogeneous group of choroidoretinal dystrophies.


[Study on the correlation among quantification of HBV-DNA and HBeAg, anti-HBe in hepatitis B carriers].
To better understand the duplication of hepatitis B virus (HBV) in order to improve clinical diagnoses and treatments via quantitative measurement of HBV-DNA and comparison of correlation of HBV-DNA with HBeAg and anti-HBe. For 883 hepatitis B patients with positive HBsAg, HBV-DNA was measured by COBAS AMPLICOR HBV MONITOR reagent and COBAS AMPLICOR quantitative PCR instrument. Microparticle enzyme immunoassay analysis (MEIA) was then carried out with fully automatic enzyme immunoassay analysis instrument made by Abbott Axsym from the U.S. to measure HBeAg and anti-HBe. Correlation was analysed by SPSS. (1)Positive correlation between 690 HBV-DNA positive and HBeAg positive with r= 0. 505 (P< 0.01) was found with mean values as:HBV-DNA:7.12 x 10(12) copies/ml;HBeAg:218.31 S/CO. HBV-DNA:10(4) copies/ml, HBeAg: 104 S/CO; HBV-DNA: 10(5)-10(8) copies/ml, HBeAg: 112 S/CO; HBV-DNA: 10(9)-10(15) copies/ml, HBeAg: 252 S/CO. (2) No correlation was found between 193 HBV-DNA and anti-HBe + with r= -0.052(P= 0.477> 0.05) with Mean: HBV-DNA: 8.0x 10(10) copies/ml anti-HBe: 0.18 S/CO. HBV-DNA and HBeAg appeared to have had linear correlation, showing that HBeAg> 100 S/CO,HBV-DNA> 10(4) copies/ml and hepatitis B virus were reproduced. However, HBV-DNA did not show linear correlation with anti-HBe as HBeAg negative and anti-HBe positive, the level of hepatitis B viral replication decrease slightly. But the virus load is still high. Infectivity can not neglect.


Richter's transformation of lymphoma complicating Gaucher's disease.
A case of lymphoma in a middle-aged woman with Gaucher's disease, a mu-kappa monoclonal gammopathy, peripheral lymphocytosis, and massive splenomegaly is described. There was no evidence of Gaucher's disease in the splenectomy specimen. Instead, the splenic architecture was effaced by a small lymphocytic lymphoma (SLL), with a few foci of large cell lymphoma (LCL). Immunoperoxidase studies showed that both lymphomatous components had a mu-kappa immunophenotype. Flow cytometric analysis of peripheral blood and spleen lymphocyte surface markers confirmed monoclonality. The intensity of surface immunoglobulin expression and the results of other cell marker studies in the SLL component were consistent with a stage of differentiation beyond that typical of chronic lymphocytic leukemia (CLL). The presence of abundant cytoplasmic immunoglobulin in the SLL and the association with monoclonal gammopathy were more consistent with Waldenström's macroglobulinemia or plasmacytoid lymphocytic lymphoma. The immunohistochemical and flow cytometric findings suggest that the SLL component evolved clonally or underwent Richter's transformation to LCL. Although Gaucher's disease has been associated with a variety of B-cell disorders, the subsequent transformation to a high-grade lymphoma has not been previously reported.


Nicotinamide relaxes vascular smooth muscle by inhibiting myosin light chain kinase-dependent signaling pathways: implications for anticancer efficacy.
Nicotinamide has been shown to be an effective tumor oxygenator in preclinical studies and is part of a promising clinical protocol for the treatment of cancer of the larynx. It has been known for some time that nicotinamide sensitizes tumors, at least in part, by modulating vascular smooth muscle contraction; however, the cellular target within the smooth muscle cell has yet to be identified. Our previous studies have eliminated targets within several agonist and antagonist signaling pathways in smooth muscle, suggesting that it must act at a point close to the contractile machinery of the cell. The present study investigated the effect of nicotinamide on four key steps responsible for force generation via actin/myosin interaction in the smooth muscle cell: calcium binding to calmodulin, calcium-calmodulin binding to smooth muscle myosin light chain kinase (MLCK) inhibitor peptide 480-501 (smMLCIP), modulation of MLCK-dependent signaling, and MLCK-induced phosphorylation of the regulatory myosin light chain, MLC20. Nicotinamide abolished the phosphorylation of MLC20, but had no significant effect on the other endpoints tested. We conclude that the vasorelaxant effects of nicotinamide are mediated mainly through inhibition of MLC20 phosphorylation, and that this could be a promising target for the development of novel tumor oxygenators to enhance radio- and chemotherapy.


[hsCRP protein in children and adolescents with diabetes type 1].
HsCRP protein is known as a novel marker of low grade inflammatory state, which characterises an atherosclerotic process in its early stages. Contrary to a large amount of data on inflammatory markers in diabetes type 2 and metabolic syndrome in adults, little is known so far about the inflammatory process in diabetes type 1, especially in children. The aim of the study was to estimate the level of hsCRP protein in children and adolescents with diabetes type 1 depending on coexisting additional risk factors for atherosclerosis and microvascular complications. 127 children and adolescents with diabetes duration 6.7+/-3.3 years, aged 14.9+/-3.1, were studied. The control group consisted of 52 healthy children aged 14.9+/-2.8 years, matched acc. to gender. HsCRP level was assessed with use of immunoturbidymetric, latex augmented method (Tina-quant CRP (Latex) HS, Roche). HsCRP in the whole study group was nearly significantly higher compared to control group: 0.17+/-0.2 vs. 0.078+/-0.1 mg/dl, p=0.072. In diabetic hypertensive children (n=38) we found significantly higher levels of hsCRP compared to controls (0.27+/-0.3 vs. 0.07 mg/dl, p=0.008) and compared to diabetic normotensive children (0.13+/-0.22 mg/dl; p=0.024). Diabetic obese patients (n=23) had significantly higer hsCRP compared to controls (0.24+/-0.3 vs. 0.07+/-0.1 mg/dl, p=0.04). In 14 studied diabetic children we found coexisting hypertension and obesity, and we found further increase in hsCRP level - 0.28+/-0.3 mg/dl. In diabetic children with microangiopathy hsCRP level was 0.22+/-0.2 mg/dl, and it was insignificantly higher compared to controls and to diabetic children without complications. Correlation analysis showed interrelations between hsCRP and systolic blood pressure (r=0.2; p=0.04) and HbA1c (r=0.25; p=0.015). In stepwise regression analysis hsCRP was related to systolic blood pressure, HbA1c and the triglycerides level (R=0.37; p=0.003). In children and adolescents with diabetes type 1 we proved significantly higher levels of hsCRP in case of a coexistence of hypertension and/or obesity. Elevated hsCRP in children with diabetes type 1 and hypertension and/or obesity reflects low grade inflammatory state in the course of metabolic syndrome.


Regulatory Themes and Variations by the Stress-Signaling Nucleotide Alarmones (p)ppGpp in Bacteria.
Bacterial stress-signaling alarmones are important components of a protective network against diverse stresses such as nutrient starvation and antibiotic assault. pppGpp and ppGpp, collectively (p)ppGpp, have well-documented regulatory roles in gene expression and protein translation. Recent work has highlighted another key function of (p)ppGpp: inducing rapid and coordinated changes in cellular metabolism by regulating enzymatic activities, especially those involved in purine nucleotide synthesis. Failure of metabolic regulation by (p)ppGpp results in the loss of coordination between metabolic and macromolecular processes, leading to cellular toxicity. In this review, we document how (p)ppGpp and newly characterized nucleotides pGpp and (p)ppApp directly regulate these enzymatic targets for metabolic remodeling. We examine targets' common determinants for alarmone interaction as well as their evolutionary diversification. We highlight classical and emerging themes in nucleotide signaling, including oligomerization and allostery along with metabolic interconversion and crosstalk, illustrating how they allow optimized bacterial adaptation to their environmental niches.


Treatment of Carotid Ophthalmic Aneurysm With Woven EndoBridge (WEB SL): 2-Dimensional Operative Video.
Wide-necked cerebral aneurysms often require complex treatment strategies for optimal treatment. As the surgeon's arsenal continues to expand, consideration of all potential applications of available devices is important. The Woven EndoBridge (WEB) device (MicroVention-Terumo, Aliso Viejo, California) capitalizes on flow disruption to promote thrombosis and is Food and Drug Administration (FDA) approved for saccular wide-necked bifurcation aneurysms located at the middle cerebral artery bifurcation, internal carotid artery (ICA) terminus, anterior communicating artery complex, and basilar apex. In this video, we demonstrate an off-label use of the WEB to treat a wide-necked type II1 ophthalmic artery aneurysm, highlighting the importance of correct device sizing. The patient is a 74-yr-old woman with a family history of aneurysms. Her aneurysm was found incidentally after a minor trauma. Observation and various treatment options were considered. The patient preferred to avoid open surgical intervention and dual antiplatelet therapy. Endoluminal flow diversion for types II and III ophthalmic artery aneurysms has relatively low occlusion rates and a higher incidence of visual field deficits.1 A WEB device can be an excellent alternative to treat these aneurysms. A biaxial system was used to selectively catheterize the supraclinoid internal carotid artery and then the aneurysm, and optimal flow diversion was achieved. The patient did well and was discharged home the next day on aspirin alone. Six-month angiography showed near-occlusion of the aneurysm and ophthalmic artery patency. The neck remnant will be followed up with repeat angiography in 6 mo. The patient gave informed consent for the procedure and video recording. Institutional review board approval was deemed unnecessary. Video ©University at Buffalo Neurosurgery, 2019. With permission.


Orphan nuclear hormone receptor Rev-erbalpha regulates the human apolipoprotein CIII promoter.
Apolipoprotein CIII (apoCIII) plays an important role in plasma triglyceride and remnant lipoprotein metabolism. Because hypertriglyceridemia is an independent risk factor in coronary artery disease and the presence in plasma of triglyceride-rich remnant lipoproteins is correlated with atherosclerosis, considerable research efforts have been focused on the identification of factors regulating apoCIII gene expression to decrease its production. Here we report that the orphan nuclear hormone receptor Rev-erbalpha regulates the human apoCIII gene promoter. In apoCIII expressing human hepatic HepG2 cells, transfection of Rev-erbalpha specifically repressed apoCIII gene promoter activity. We determined by deletion and site-directed mutagenesis experiments that Rev-erbalpha dependent repression is mainly due to an element present in the proximal promoter of the apoCIII gene. In contrast, we found no functional Rev-erbalpha response elements in the convergently transcribed human apoAI gene or the common regulatory enhancer. The identified Rev-erbalpha response element coincides with a RORalpha1 element, and in the present study we provide evidence that functional cross-talk between these orphan receptors modulates the apoCIII promoter. In vitro binding analysis showed that monomers of Rev-erbalpha bound this element but not another upstream RORalpha1 response element. In addition, we showed that the closely related nuclear orphan receptor RVR also specifically repressed the human apoCIII gene. These studies underscore a novel physiological role for members of the Rev-erb family of nuclear receptors in the regulation of genes involved in triglyceride metabolism and the pathogenesis of atherosclerosis.


A root-specific wall-associated kinase gene, HvWAK1, regulates root growth and is highly divergent in barley and other cereals.
Wall-associated receptor-like kinases (WAKs) are important candidates for directly linking the extracellular matrix with intracellular compartments and are involved in developmental processes and stress response. WAK gene family has been identified in plants such as Arabidopsis and rice. Here, we present a detailed analysis of the WAK1 gene from barley cv. Golden Promise, mapped to chromosome 5H. Three BAC clones corresponding to the WAK fragment were sequenced and the full-length WAK1 gene was characterized. The gene has three exons and two short introns with a coding region of 2,178 bp encoding a protein of 725 amino acids. A regulatory region was analyzed in -1,000 bp sequence upstream to start codon. Using conserved domains database and SMART, various conserved domains such as GUB WAK Bind, epidermal growth factor CA, and protein kinase C as well as other regions like signal peptides, active sites, and transmembrane domains were identified. The gene organization of HvWAK1 was compared with wheat (TaWAK1) and Arabidopsis (AtWAK1), suggesting that the WAK1 gene organization has remained highly conserved. Nonetheless, WAK1 was found to be highly divergent when compared with sequences available from barley cv. Haruna Nijo (50 %), rice (46 %), wheat (21 %), Arabidopsis (25 %), and maize (19 %). This divergence may have facilitated a better adaptation to surrounding environments due to its role in communication between the extracellular matrix, cell, and outer environment. Semiquantitative RT-PCR-based expression analysis indicates HvWAK1 expression is specific to roots. Significant differences in root growth between GP wild type and GP-Ds mutant seedlings were observed under control and salt stress conditions.


A simulation laboratory for medical education.
Simulation as an instructional technique is being utilized for the learning of clinical skills in the training of health professionals. The Center for Educational Development at the University of Illinois College of Medicine has established a Learning Resource and Development Center to assist medical faculty in the development and evaluation of instructional simulations. These are not designed to replace clinical experiences but to provide additional opportunities for students to strengthen skills in patient examination. Prior to the introduction of these learning tools in 1972, a study was undertaken to validate their instructional effectiveness. After an additional two years of operation and experimentation, a revalidation study was undertaken during the fall of 1974. Improvements are continually being made in the various simulation formats as a result of feedback from students and faculty.


Unconscious information integration: A replication and the role of spatial window in masking experiments.
Consciousness and high-level information integration have commonly been closely related to each other (Baars, 2002; Dehaene & Naccache, 2001; Tononi, 2004). Different results, however, have challenged this assumption by showing that information integration can occur for stimuli presented outside of conscious awareness. More recently, a re-examination of some of the data and different replication attempts questioned these results thereby again suggesting a close link between consciousness and information integration. The current study aimed at (i) replicating another piece of evidence for unconscious information integration and (ii) investigating if the size of the spatial window in which the information to be integrated is presented could explain why unconscious information integration sometimes fails. Results showed a reliable replication so providing further evidence for unconscious information integration in a subliminal priming paradigm. Furthermore, our results revealed that unconscious integration depends on the size of the spatial window in which the information is presented.


Duodenal erosions, a common and distinctive feature of portal hypertensive duodenopathy.
The aim of the present study was to assess the presence of duodenal erosion and its clinical characteristics on endoscopy in patients with portal hypertension who had undergone endoscopic injection sclerotherapy and/or endoscopic variceal ligation for esophagogastric varices. The subjects were 440 patients with portal hypertension, 450 with chronic hepatitis as a related control group, and 450 who underwent upper endoscopic examination as part of their routine physical examination as the controls. The underlying hepatic disease, hepatic function, and endoscopic findings of duodenal erosion among the patients with portal hypertension were studied. Duodenal erosion was found in 68 patients with portal hypertension (68 of 440, 15.5%), four patients with chronic hepatitis (four of 450, 0.9%), and two controls (two of 450, 0.4%). The incidence of duodenal erosion among the patients with portal hypertension was significantly higher than that in the other two groups (p < 0.01, p < 0.01, respectively). The lesions commonly observed in duodenitis are speckle erosions mainly located in the duodenal bulb. However, the most frequently seen form of duodenal erosion among the patients with portal hypertension extended from the superior portion to the descending portion, and tended to show a circular alignment along the Kerckring's folds. The patients with portal hypertension with reduced hepatic reserve capacity had more severe duodenal erosion. Endoscopic ultrasonography revealed thickening of the duodenal wall and proliferation of vascular structures within and around the wall. The histological findings of the duodenal erosion included edema and vascular dilation in the mucosal and submucosal layers. The location of duodenal erosion in patients with portal hypertension differs from that in patients with ordinary duodenitis. Duodenal erosion in patients with portal hypertension is considered to be one of the lesions of portal hypertensive duodenopathy.


Grading the severity of cervical neoplasia based on combined histopathology, cytopathology, and HPV genotype distribution among 1,700 women referred to colposcopy in Oklahoma.
Diagnosis and treatment of cervical cancer precursors rely on colposcopic biopsy, which is sometimes hampered by incorrect biopsy placement and the unclear prognostic value of poorly reproducible diagnoses such as cervical intraepithelial neoplasia (CIN) Grade 1 and 2. Searching for discrete disease categories that incorporate the value of cytology and that reflect the causal role of particular HPV types, we analyzed histology, cytology and HPV genotype distributions in the Study to Understand Cervical Cancer Endpoints and Early Determinants (SUCCEED). This cross-sectional study comprises approximately 1,700 women referred to colposcopy or treatment for the spectrum of cervical disease, including 439 women with <CIN1, 429 CIN1, 322 CIN2, 297 CIN3 and 107 with cancer. Using hierarchical clustering of histology-cytology groups based on HPV genotype distributions, we could plainly distinguish in this referral population 5 increasingly severe diagnostic groups of HPV-positive women: (i) HPV-positive women with <CIN2 histology and normal cytology, (ii) HPV positive women with <CIN2 histology and ASC or LSIL cytology; (iii) CIN2, including histologic CIN2 and HSIL cytology with any histology <CIN2; (iv) CIN3; and (v) invasive cervical cancer. The grouping of women with HSIL cytology, but without histological abnormalities to women with CIN2 suggests that in these cases the worst lesion was missed during colposcopy-biopsy. We are now using these sharpened diagnostic categories to search for novel biomarkers predicting the risk of progression and invasion.


On the contribution of S100A10 and annexin A2 to plasminogen activation and oncogenesis: an enduring ambiguity.
Plasminogen receptors are becoming increasingly relevant in regulating many diseases such as cancer, stroke and inflammation. However, controversy has emerged concerning the putative role of some receptors, in particular annexin A2, in binding plasminogen. Several reports failed to account for the effects of annexin A2 on the stability and conformation of its binding partner S100A10. This has created an enduring ambiguity as to the actual function of annexin A2 in plasmin regulation. Supported by a long line of evidence, we conclude that S100A10, and not annexin A2, is the primary plasminogen receptor within the annexin A2-S100A10 complex and contributes to the plasmin-mediated effects that were originally ascribed to annexin A2.


Comparison of polyetheretherketone cages with femoral cortical bone allograft as a single-piece interbody spacer in transforaminal lumbar interbody fusion.
Transforaminal lumbar interbody fusion (TLIF) is an accepted alternative to circumferential fusion of the lumbar spine in the treatment of degenerative disc disease, spondylolisthesis, and recurrent disc herniation. To maintain disc height while arthrodesis takes place, the technique requires the use of an interbody spacer. Although titanium cages are used in this capacity, the two most common spacers are polyetheretherketone (PEEK) cages and femoral cortical allografts (FCAs). The authors compared the clinical and radiographic outcomes of patients who underwent TLIF with pedicle screw fixation, in whom either a PEEK cage or an FCA was placed as an interbody spacer. The charts and x-ray films obtained in 39 patients (age range 33-68 years, mean 44.7 years) who underwent single-level TLIF between October 2001 and April 2004 and in whom either a PEEK cage (18 patients) or FCA (21 patients) was placed as an interbody spacer were evaluated in a retrospective study. Radiological outcome was based on fusion rate and a comparison of the initial postoperative lordotic angle on standing lateral radiographs with that at long-term follow up (mean follow up 15.1 months, minimum 12 months). To control for variations in radiographic magnification, the authors used lordotic angle as an indirect measure of disc space height. Clinical outcome was assessed using the Oswestry Disability Index (ODI). There were no major complications in either group. Radiographically documented fusion occurred in all patients in the PEEK group and 95.2% of those in the FCA group. Pseudarthrosis developed in one patient in the FCA group, and this patient underwent additional surgery. In both groups, the mean lordotic angle changed by less than 2.20 degrees during the postoperative period, and the mean postoperative ODI score was more than 40 points lower than the mean preoperative score. There was no significant difference between the two groups in mean change in lordotic angle (p = 0.415) and mean change in ODI score (p = 0.491). Both PEEK cages and FCAs are highly effective in promoting interbody fusion, maintaining postoperative disc space height, and achieving desirable clinical outcomes in patients who undergo TLIF with pedicle screw fixation. The advantages of PEEK cages include a lower incidence of subsidence and their radiolucency, which permits easier visualization of bone growth.


Synapse Elimination Triggered by BMP4 Exocytosis and Presynaptic BMP Receptor Activation.
In vitro screening of signaling molecules involved in neural circuit formation has identified a large number of synaptogenic proteins. However, factors that drive synapse elimination remain elusive. Here, we report that bone morphogenetic protein 4 (BMP4) released from axons has the ability to eliminate synapses. We found fast axonal transport of BMP4 in dense-core vesicles, its exocytosis, and subsequent cell surface clustering via type I BMP receptors near synapses. BMP4 overexpression or knockout in culture reduced or increased presynaptic structures, respectively. The destabilizing effect of surface BMP4 clusters was limited to nearby synapses. In vivo knockout of BMP4 and subsequent two-photon imaging of synapse dynamics confirmed its critical role in maintaining an appropriate density of presynaptic components along the axon. These results suggest an essential role for perisynaptic clustering of BMP4 during development in the construction of functional neuronal circuits.


Expression of microRNA-1 and microRNA-21 in different protocols of ischemic conditioning in an isolated rat heart model.
'Conditioning' [ischemic preconditioning (IPC), ischemic postconditioning (IPO) and remote ischemic preconditioning (RIPC)] the heart to render it more resistant to an episode of acute myocardial ischemia-reperfusion (I/R) injury is an endogenous cardioprotective strategy. There are several mechanisms proposed for 'conditioning', such as endogenous mediators or cytoprotective proteins. In recent reports, microRNAs (miRNAs) were involved in controlling the expression of myocardial ischemia-related genes. Some studies have demonstrated that cardiac miRNA-1 and miRNA-21 were significantly increased by late IPC with an increase in their target proteins [endothelial nitric oxide synthase and heat shock protein 70 (HSP70)], but their expression levels in 'conditioning' strategies are currently unknown. In the current study, Langendorff-perfused Sprague-Dawley rat hearts were randomly assigned to one of four groups [control group (CON group, n = 12), IPC group (n = 12), IPO group (n = 12) and RIPC group (n = 12)]. Cardiac function was digitalized and analyzed. The expression of miRNA-1 and miRNA-21 was detected by real-time reverse transcription polymerase chain reaction. The expression of HSP70, programmed cell death protein 4 (PDCD4), B-cell lymphoma/leukemia-2 (Bcl-2) and Bcl-2-associated X protein (Bax) was detected by Western blot. Cardiac infarct size and myocardial apoptosis were determined using the 2,3,5-triphenyltetrazolium chloride assay and terminal deoxynucleotidyl transferase dUTP nick end labeling assay, respectively. The results revealed that miRNA-1 (233 ± 45%) and miRNA-21 (356 ± 33%) expression was up-regulated in the IPC group, but the expression of miRNA-1 was down-regulated in the RIPC (61 ± 16%) group and IPO group (61 ± 13%). The expression of PDCD4 [IPC (74 ± 11%), RIPC (81 ± 16%), IPO (83 ± 12%)], HSP70 [IPC (74 ± 5%), RIPC (81 ± 6%), IPO (67 ± 11%)] and Bax [IPC (27 ± 6%), RIPC (21 ± 3%), IPO (27 ± 4%)] was down-regulated in the conditioning groups compared with the CON group [PDCD4 (130 ± 11%), HSP70 (121 ± 11%) and Bax (63 ± 8%)]. In the conditioning hearts, infarct size [IPC (31.7 ± 4.1%), RIPC (29.6 ± 6.19%) and IPO (32.8 ± 4.71%)] and myocardial apoptosis [IPC (15.2 ± 4.21%), RIPC (17.2 ± 1.92%) and IPO (15.6 ± 4.04%)] were significantly decreased compared with the CON group (infarct size: 51.77 ± 4.3%, myocardial apoptosis: 32.8 ± 3.96%). We concluded that miRNA-1 and miRNA-21 expression differed in IPC, RIPC and IPO groups, and their target proteins were not inversely correlated with the miRNAs in all the conditioning groups, which revealed that the miRNAs were regulated but complicated by the different conditioning protocols.


mRNA cap regulation in mammalian cell function and fate.
In this review we explore the regulation of mRNA cap formation and its impact on mammalian cells. The mRNA cap is a highly methylated modification of the 5' end of RNA pol II-transcribed RNA. It protects RNA from degradation, recruits complexes involved in RNA processing, export and translation initiation, and marks cellular mRNA as "self" to avoid recognition by the innate immune system. The mRNA cap can be viewed as a unique mark which selects RNA pol II transcripts for specific processing and translation. Over recent years, examples of regulation of mRNA cap formation have emerged, induced by oncogenes, developmental pathways and during the cell cycle. These signalling pathways regulate the rate and extent of mRNA cap formation, resulting in changes in gene expression, cell physiology and cell function.


Efficacy of the first oral vaccination against fox rabies in Slovenia.
Fox population reduction was the first measure undertaken to control rabies in foxes, but this proved unsuccessful. The promising results of oral immunisation of foxes against rabies in some European countries encouraged the authorization of the first rabies vaccination of foxes in the field in Slovenia, in October 1988. In the present study, intervention analysis is used to evaluate the results of this vaccination campaign. The analysis took into account the cyclic nature of fox rabies and the possible effects of variability in the fox carcass submission rate. The results confirmed the decrease in fox rabies after the launch of the vaccination campaign. The reduction was independent of both cyclic oscillations in fox rabies and variability in carcass submission rate, thus indicating a positive net effect of the vaccination.


Intestinal Spirochetosis: Case Series and Review of the Literature.
This study aims to present the clinical, endoscopic, and histopathologic characteristics associated with intestinal spirochetosis (IS). It also serves to heighten awareness among pathologists, since the histologic appearance of spirochetosis could be subtle and easily overlooked. Hematoxylin & eosin (H&E) slides and special stains of intestinal biopsies from six patients with a diagnosis of IS at our institution were reviewed. Clinical history, endoscopic, and histopathologic findings were obtained from electronic medical records. The patients presented with diverse clinical symptoms, and only one patient was asymptomatic. The most consistent symptoms were watery diarrhea and abdominal cramps. Two out of five treated patients reported symptomatic improvement after antibiotics therapy. The colonoscopy findings were not specific, ranging from normal mucosa to polyps, to mucosal ulcerations in one patient. On histologic examination, the typical "brush-like" organisms lying perpendicular to the surface epithelium are seen both on H&E stain and special stains. IS is usually an incidental histologic finding, and the association with symptoms is still unclear. The clinical presentation could be very diverse, hence, a long list of differential diagnosis should be ruled out. Additional clinical testing should be pursued if patients are unresponsive to antibiotic treatment.


Enhancement of the second-harmonic generation in a quantum dot-metallic nanoparticle hybrid system.
We have investigated the second-harmonic generation (SHG) and dipole-dipole interaction in a quantum dot and metallic nanoparticle hybrid system. A strong probe field is applied to create two-photon absorption in the quantum dot and metallic nanoparticle. SHG photons and SHG surface plasmon polaritons are emitted by the quantum dot and metallic nanoparticle, respectively. Induced dipoles are created in the quantum dot and the metallic nanoparticle due to two-photon absorption and hence both systems interact with each other via the dipole-dipole interaction. It is found that SHG signals produced by the quantum dot and nanoparticle are enhanced by the dipole-dipole interaction and also that the SHG signal can be switched on and off by applying a control field. The theoretical findings of this paper are supported by recent experimental studies. The present hybrid system can be used to fabricate nano-sensors and all-optical nano-switching devices.


Preliminary recovery study of a commercial molecularly imprinted polymer for the extraction of glyphosate and AMPA in different environmental waters using MS.
A commercial molecularly imprinted polymer (MIP) dedicated to glyphosate (GLY) and its main metabolite, aminomethylphosphonic acid (AMPA), was lately assessed as "POCIS-like" sampler on mineral water. The obtained results were encouraging with 111 and 122 mL day-1 as sampling rates for GLY and AMPA, respectively. Therefore, before applying this passive sampler to environmental waters, the commercial phase was tested on different water matrices as a solid-phase extraction (SPE) device. The SPE protocol was carried on 250 mg of MIP with the following three steps: conditioning by Milli-Q water, loading of the sample (15 mL), and elution of the analytes by 4 mL 0.1 M HCl that were evaporated to dryness and recovered in 15 mL of the suitable solvent for analysis. This protocol was first applied to mineral water spiked by GLY and AMPA at environmental concentration levels (25-750 ng L-1). Analyses were carried out by ultra-performance liquid chromatography hyphenated to tandem mass after derivatization of GLY and AMPA by 9-fluorenylmethylchloroformate. The linear correlation between concentrations measured with and without SPE on MIP was proved.Furthermore, other extractions showed that high concentrations of metal ion interferents (lead(II), cadmium(II), and zinc(II)) in groundwaters did not reduce SPE performance of the MIP.Then, concentration assays were undertaken and brought noteworthy results, such as the recovery of 80% GLY and AMPA from groundwater spiked at 10 ng L-1 and concentrated 100 times. For this purpose, ion exclusion chromatography hyphenated to mass was applied without previous derivatization of the analytes. The same concentration factor and analytical method were applied to 100 ng L-1 spiked sea water with recoveries of 96% for GLY and 121% for AMPA.


[Comparative study of surgical effects on patients with mixed deafness and otosclerosis with different air bone conduction differences].
Objective: To analyze the surgical efficacy of patients with mixed hearing loss and otosclerosis with different air bone gap (ABG) before surgery, and to provide reference for the prognosis evaluation of otosclerosis surgery. Methods: The clinical data of 108 cases(116 ears) of otosclerosis who had undergone stapes fenestration technique artificial stapes implantation in Xiangya Hospital of Central South University from November 2013 to May 2020 and had mixed hearing loss before surgery were collected, including 71 women(76 ears)and 37 men (40 ears), with an average age of 38.5 years. According to preoperative pure tone audiometry ABG, they were divided into three groups: group S, 15 dB≤ABG<31 dB, a total of 39 ears; group M, 31 dB≤ABG<46 dB, a total of 58 ears; and group L, ABG≥46 dB, 19 ears in total. The hearing outcomes of three groups of patients at 6-12 months after surgery were compared and analyzed using SPSS 24.0 statistical software. Results: A total of 3 patients (group S: 2 cases; group L: 1 case) experienced severe sensorineural hearing loss after surgery and were not included in the statistical analysis. After surgery, the pure tone hearing threshold of patients with otosclerosis in each group was significantly improved compared to before surgery, with an average air conduction threshold improvement of(21.6±13.4) dB. The difference between before and after surgery was statistically significant(t=17.13, P<0.01). The average bone conduction threshold improved by(3.7±7.6) dB, and the difference was statistically significant before and after surgery(t=5.20, P<0.01). The postoperative ABG was(18.3±9.3) dB, which was significantly reduced compared to preoperative(36.2±8.6)dB. Among the three groups of patients, the L group had the highest improvement in air conduction threshold[(29.9±10.8)dB], while the S group had the lowest improvement[(15.7±11.4)dB]. There was no statistically significant difference in post operative pure tone hearing thresholds between the three groups(P>0.05). The postoperative ABG in group S was the smallest[(16.5±9.0)dB], while in group L, the postoperative ABG was the largest[(20.5±10.0)dB]. Compared with group S, group M and group L still had a large residual ABG at 2 000 Hz after surgery. The bone conduction threshold of both S and M groups improved to some extent after surgery compared to before (P<0.01). Conclusions: Surgery can benefit patients with mixed hearing loss and otosclerosis with different preoperative ABG. Patients with small preoperative ABG have better surgical results and ideal ABG closure at all frequencies after surgery. Patients with large preoperative ABG can significantly increase the gas conduction threshold during surgery, but certain frequencies of ABG may still be left behind after surgery. The improvement effect of surgery on bone conduction threshold is not significant. Patients should be informed of treatment methods such as hearing aids based on their actual situation for selection.


A highly selective fluorescent chemosensor for lead ions.
A simple fluorescent chemosensor 1 exhibits a 40-fold fluorescence enhancement upon addition of lead ion and yet is insensitive to most of univalent and divalent metal ions. The high affinity of 1 toward Pb2+ presumably results from the cooperative binding behaviors of two potential metal binding sites: the dicarbonyl and the 15-monoazacrown-5 ether.


Some properties of purified phospholipase D and especially the effect of amphipathic substances.
1. The soluble phospholipase D of cabbage was purified by heat treatment, acetone precipitation and electrophoresis on a density gradient of aqueous glycerol. 2. The purified enzyme slowly attacked a lecithin suspension whereas ultrasonically treated lecithin was hydrolysed more rapidly. 3. Diethyl ether stimulated the hydrolysis of both the lecithin suspension and ultrasonically treated lecithin. 4. Ca(2+) was essential for the hydrolysis (optimum about 0.04m); it could not be replaced by Mg(2+) or cationic amphipathic substances. 5. The reaction had a sharp pH optimum at pH5.4, irrespective of the physical form of the lecithin substrate or the activator used. 6. Anionic amphipathic substances such as dodecyl sulphate, phosphatidic acid, triphosphoinositide and monocetyl phosphoric acid, were potent activators of the reaction: other acidic lipids were relatively inactive. 7. Cationic amphipathic substances inhibited the hydrolysis; however, they also reversed the inhibition caused by using an excess of anionic amphipathic substance as activator. 8. The activation produced by amphipathic substances could not be correlated with their effect on the zeta-potential or size of the substrate particles. 9. The addition of activating anionic amphipaths to lecithin induces the latter to adsorb enzyme from solution. In the absence of Ca(2+) the enzyme is denatured on the highly negatively charged surface, but in the presence of Ca(2+) (or Mg(2+)) it is protected from denaturation. It is suggested that this adsorption is an essential prerequisite for ready enzymic hydrolysis. 10. The hydrolysis of lecithin by the enzyme was strongly inhibited by protamine sulphate (0.1mg./ml.) and by choline and ethanolamine. 11. Ultrasonically treated phosphatidylethanolamine, or mixed particles of phosphatidylethanolamine plus dodecyl sulphate, were slowly attacked by the enzyme provided that Ca(2+) was present.


Chiral separation and determination of amino acid enantiomers in fruit juice by open-tubular nano liquid chromatography.
A novel chiral porous-layer stationary phase was developed for use in open-tubular nano liquid chromatography. The stationary phase was prepared by an in-situ polymerization of 3-chloro-2-hydroxypropylmethacrylate (HPMA-Cl) and ethylene dimethacrylate (EDMA). The reactive chloro groups at the surface of the porous stationary phase were reacted with β-Cyclodextrin (β-CD). The resulting morphology was characterized by using scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FT-IR). The chromatographic performance of the column was evaluated by hydrophilic interaction chromatography (HILIC). Amino acids were used as test solutes. The running buffer conditions for the enantioseparation were found to be 85% acetonitrile (ACN):10%MeOH: 5% H2 O at 0.1% v/v trifluoro acetic acid (TFA) (flow rate: 800 nL/min). The enantioseparation provided high theoretical plate numbers up to 26 000 platesm-1 . A good retention capacity within satisfactory retention times was also achieved. Real sample applicability of this column to the separation of amino acid enantiomers in fruit juice sample was demonstrated.


Response of psoriatic lesions to multiple applications of leukotriene B4 and 12-HETE.
Single topical application of the neutrophil chemoattractant arachidonic acid metabolites leukotriene B4 (LTB4) and 12-R,S-hydroxy-5,8,10,14-eicosatetraenoic acid [corrected] (12-HETE) to normal skin elicits a histological response similar to early psoriasis. This effect diminishes following repeated application indicating the development of local tolerance. In order to assess whether induction of tolerance could be exploited as a treatment for psoriasis we studied the clinical effects of topical application of LTB4 (n = 6) and 12-HETE (n = 3) under occlusion to small psoriatic lesions for 12 consecutive days. The area of the control lesions decreased significantly over the study period while the areas of lesions to which LTB4 and 12-HETE were applied remained unchanged. No other difference between responses of the three groups was detected. We have shown that, in the doses used, multiple applications of these substances to established stable psoriatic lesions do not produce a therapeutically useful response.


New components of the Dictyostelium PKA pathway revealed by Bayesian analysis of expression data.
Identifying candidate genes in genetic networks is important for understanding regulation and biological function. Large gene expression datasets contain relevant information about genetic networks, but mining the data is not a trivial task. Algorithms that infer Bayesian networks from expression data are powerful tools for learning complex genetic networks, since they can incorporate prior knowledge and uncover higher-order dependencies among genes. However, these algorithms are computationally demanding, so novel techniques that allow targeted exploration for discovering new members of known pathways are essential. Here we describe a Bayesian network approach that addresses a specific network within a large dataset to discover new components. Our algorithm draws individual genes from a large gene-expression repository, and ranks them as potential members of a known pathway. We apply this method to discover new components of the cAMP-dependent protein kinase (PKA) pathway, a central regulator of Dictyostelium discoideum development. The PKA network is well studied in D. discoideum but the transcriptional networks that regulate PKA activity and the transcriptional outcomes of PKA function are largely unknown. Most of the genes highly ranked by our method encode either known components of the PKA pathway or are good candidates. We tested 5 uncharacterized highly ranked genes by creating mutant strains and identified a candidate cAMP-response element-binding protein, yet undiscovered in D. discoideum, and a histidine kinase, a candidate upstream regulator of PKA activity. The single-gene expansion method is useful in identifying new components of known pathways. The method takes advantage of the Bayesian framework to incorporate prior biological knowledge and discovers higher-order dependencies among genes while greatly reducing the computational resources required to process high-throughput datasets.


Integrated expression of glomerular extracellular matrix proteins and beta 1 integrins in monoclonal light chain-related renal diseases.
Light-chain deposition disease (LCDD) and amyloid light-chain amyloidosis (AL-Am) represent the two classical diseases associated with glomerular alterations in monoclonal light chain-related renal diseases. LCDD is characterized by deposition of extracellular matrix proteins in the mesangium, thus creating the morphologic appearance recognized as nodular glomerulosclerosis. In AL-Am, the mesangial matrix is replaced by polymerized light chains in the form of amyloid fibrils. Integrins are responsible for cell-to-cell and cell-to-matrix communication and, therefore, are expected to play a key role in the alterations encountered in these two diseases. The present article addresses the expression of selected extracellular matrix proteins (collagen IV, laminin, fibronectin, and tenascin) and their respective receptor beta 1 integrins (alpha 2, alpha 3, alpha 5, and alpha 9) in glomeruli with LCDD and AL-Am by immunohistochemical methods. The corresponding integrin (alpha 9 beta 1) co-localized with tenascin in the center of the mesangial nodules in LCDD. In AL-Am, tenascin is found primarily at the periphery of replaced mesangial areas and in the remaining mesangium not replaced by the amyloid. Tenascin co-localized with alpha 9 beta 1 integrin in mesangial areas in the earlier phases of the process. Fibronectin, laminin, and collagen IV, although increased in absolute amounts, are pushed toward the periphery of mesangial areas, in which correlated expression of their corresponding beta 1 integrins (alpha 2, alpha 3, and alpha 5, respectively) is documented in both LCDD and AL-Am. Deposition of tenascin might be at least partially responsible for the perpetuation and irreversibility of the glomerular lesion in LCDD.


Production of lymphokine-activated lymphocytes. Lysis of human head and neck squamous cell carcinoma cell lines.
Lymphokine-activated killer cells are thought to be important mediators of host tumor defense. In the present study, the cytotoxic potential of lymphokine-activated lymphocytes against different head and neck squamous cell carcinoma cell lines was investigated. Lymphokine-activated killer cells were derived from peripheral blood lymphocytes. Effector peripheral blood lymphocyte cell suspensions were incubated in the presence or absence of recombinant interleukin-2. Cytotoxicity of incubated cells or fresh peripheral blood lymphocytes was determined in a 3-hour chromium 51 release assay. Target cell lines included K562 (a natural killer-sensitive target) and the following head and neck squamous cell carcinoma cell lines: Cal 27, UMSCC-1, UMSCC-8, UMSCC-16, UMSCC-19, and UMSCC-22a. Fresh peripheral blood lymphocytes and peripheral blood lymphocytes cultured in the absence of added interleukin-2 demonstrated minimal cytotoxic effects against the squamous cell carcinoma targets. In contrast, these fresh and incubated lymphocytes showed significant cytotoxic effects against K562. Cells preincubated in the presence of interleukin-2 demonstrated a statistically significant increase in cytotoxic effects against K562 and all squamous cell carcinoma targets. These investigations support the possible role of lymphokine-activated killer cells in host defense against squamous cell carcinoma. In vitro natural killer cell activity against head and neck squamous cell carcinoma cell lines is low; however, significant lymphokine-activated killer cell cytotoxicity is present.


[Extraintestinal infections caused by Vibrio parahaemolyticus and Vibrio alginolyticus at the county of Funen 1987-1992].
Over a six-year period we detected 30 clinical infections caused by halophilic vibrios in a restricted geographical area. Vibrio parahaemolyticus infections were found in 13 patients (three with wound infections, ten with ear infections), and Vibrio alginolyticus infections in 17 patients, all of whom had ear infections. From 1987 to 1990, infections caused by marine vibrios were found in at most four cases annually, in 1992 in six instances, whereas over a five-month period in 1991 we experienced 15 cases of extraintestinal infections. Fifteen of the infections found in 1991 and 1992 were ear infections, ten of which occurred shortly after exposure to Danish coastal seawater. The disease was mild in most of the patients and all recovered. Most of the patients showed predisposing conditions such as chronic otitis media, perforation of the tympanic membrane or ulcus cruris. The organisms were isolated in minor numbers from coastal water samples from five of 12 bathing areas around Funen at the end of July, but not at the end of November. This study indicates that halophilic marine vibrios may be pathogenic in Denmark in persons exposed to seawater.


Increased Otoferlin Expression in B Cells Is Associated with Muscle Weakness in Untreated Juvenile Dermatomyositis: A Pilot Study.
Otoferlin mRNA expression is increased in JDM patients' PBMCs and muscle compared to healthy controls. This study aims to evaluate the role of otoferlin in JDM disease pathophysiology and its association with disease activity in untreated children with JDM. A total of 26 untreated JDM (88.5% female, 92.3% white, non-Hispanic) and 15 healthy controls were included in this study. Otoferlin mRNA expression was determined by qRT-PCR before and a few months after therapy. Detailed flow cytometry of various cell surface markers and cytoplasmic otoferlin was performed to identify cells expressing otoferlin. In addition, muscle otoferlin expression was evaluated in situ in six untreated JDM patients and three healthy controls. There was a significant increase in otoferlin expression in JDM children compared to controls (Median 67.5 vs. 2.1; p = 0.001). There was a positive correlation between mRNA otoferlin expression and the following disease activity markers: disease activity scores (DAS)-total (rs = 0.62, p < 0.001); childhood myositis assessment scale (CMAS) (rs = -0.61, p = 0.002); neopterin (rs = 0.57, p = 0.004) and von Willebrand factor antigen (vWF: Ag) (rs = 0.60, p = 0.004). Most of the otoferlin-positive cells were unswitched B cells (63-99.4%), with 65-75% of them expressing plasmablast markers (CD19+, IgM+, CD38hi, CD24-). The findings of this pilot study suggest that otoferlin expression is associated with muscle weakness, making it a possible biomarker of disease activity. Additionally, B cells and plasmablasts were the primary cells expressing otoferlin.


Effectiveness of a Labor Cervical Exam Model in Family Medicine and OB-GYN Residents.
Labor cervical exam accuracy is an essential skill for family medicine and OB-GYN residents to master. To determine the effectiveness of simulation on labor cervical exam training, family medicine and OB-GYN residents were trained using a self-constructed PVC pipe-based cervical exam model during a short and intensive simulation workshop or "boot camp." A task trainer was constructed that allows for the blind examination of cervical dilation and effacement. This model was used in the training of first-year family medicine and OB-GYN residents during an 8 day simulation course. A longitudinal comparison of pre- and post-training accuracy was performed. Using a cohort design, the post-training accuracy of first-year family medicine and OB-GYN residents (interns) was also compared to second-fourth year OB-GYN residents. Use of the model by interns (n=25) resulted in significant improvements in the accuracy of their assessments of cervical dilation, but not effacement, and decreased intra-rater variability. When compared to the second-fourth year residents (n=25) who received traditional training, but not simulation training, interns were significantly more accurate and showed less intra-rater variability in their assessments of both dilation and effacement immediately after training compared to their senior colleagues. Training with the cervical exam model improved interns' accuracy and precision immediately after an 8-day simulation course. Use of this model in resident education may aid in the early stages of training and benefit more experienced trainees by augmenting traditional clinical training.


The hows and whys of face processing: Level of construal influences the holistic processing of human faces.
Face recognition and identification are optimized by holistic processing. Various visual-spatial manipulations appear to have transfer effects on holistic face processing. The present experiment tests the effects of a semantic manipulation--of construal level--on holistic processing as measured by composite congruency effects. Participants completed two blocks of trials. The first served as a baseline, whereas the second included a manipulation of construal level. High-level construal resulted in stronger congruency effects, indicative of greater holistic processing (relative to baseline and to low-level construal). These results have implications for conceptualizations of both construal level and holistic processing.


Perception of acceptable antibiotic stewardship strategies in outpatient settings.
Antibiotics are still often inappropriately prescribed in France despite specific measures being taken for over 10years. The 25% decrease in antibiotic prescription advocated in the 2011-2016 National Antibiotic Plan seems difficult to achieve. One of the strategies currently considered in France is the use of a specific prescription form dedicated to antibiotics, with an educational message for patients. We aimed to evaluate the acceptability - by primary care prescribers - of this measure and to evaluate their perception of other antibiotic stewardship strategies. Qualitative study conducted among family physicians, pediatricians, dermatologists, dentists, and ENT specialists using semi-structured interviews. A thematic and framework analysis was then performed. Thirty prescribing physicians practicing in a specific region of France were included in the study. The dedicated prescription form for antibiotics was deemed excessive and questionable. Other measures, not directly targeting prescribers, were rather well perceived: the unit sales of antibiotics, the restricted reporting of susceptibility tests, or the limitation of the number of molecules available in outpatient settings. The results of this exploratory study may guide the national antibiotic stewardship policy in France.