mTOR Signaling Regulates Metabolic Function in Oligodendrocyte Precursor Cells and Promotes Efficient Brain Remyelination in the Cuprizone Model.
In demyelinating diseases, such as multiple sclerosis, primary loss of myelin and subsequent neuronal degeneration throughout the CNS impair patient functionality. While the importance of mechanistic target of rapamycin (mTOR) signaling during developmental myelination is known, no studies have yet directly examined the function of mTOR signaling specifically in the oligodendrocyte (OL) lineage during remyelination. Here, we conditionally deleted Mtor from adult oligodendrocyte precursor cells (OPCs) using Ng2-CreERT in male adult mice to test its function in new OLs responsible for remyelination. During early remyelination after cuprizone-induced demyelination, mice lacking mTOR in adult OPCs had unchanged OL numbers but thinner myelin. Myelin thickness recovered by late-stage repair, suggesting a delay in myelin production when Mtor is deleted from adult OPCs. Surprisingly, loss of mTOR in OPCs had no effect on efficiency of remyelination after lysophosphatidylcholine lesions in either the spinal cord or corpus callosum, suggesting that mTOR signaling functions specifically in a pathway dysregulated by cuprizone to promote remyelination efficiency. We further determined that cuprizone and inhibition of mTOR cooperatively compromise metabolic function in primary rat OLs undergoing differentiation. Together, our results support the conclusion that mTOR signaling in OPCs is required to overcome the metabolic dysfunction in the cuprizone-demyelinated adult brain.SIGNIFICANCE STATEMENT Impaired remyelination by oligodendrocytes contributes to the progressive pathology in multiple sclerosis, so it is critical to identify mechanisms of improving remyelination. The goal of this study was to examine mechanistic target of rapamycin (mTOR) signaling in remyelination. Here, we provide evidence that mTOR signaling promotes efficient remyelination of the brain after cuprizone-mediated demyelination but has no effect on remyelination after lysophosphatidylcholine demyelination in the spinal cord or brain. We also present novel data revealing that mTOR inhibition and cuprizone treatment additively affect the metabolic profile of differentiating oligodendrocytes, supporting a mechanism for the observed remyelination delay. These data suggest that altered metabolic function may underlie failure of remyelination in multiple sclerosis lesions and that mTOR signaling may be of therapeutic potential for promoting remyelination.


Effect of thymoquinone on ethylene glycol-induced kidney calculi in rats.
The aim of this study was to investigate the effects of thymoquinone, a major component of Nigella Sativa seeds on ethylene glycol-induced kidney calculi in rats. Sixty male Wistar rats were randomly divided into 6 groups (intact control, ethylene glycol control, and 4 experimental groups) and treated for 28 days according to the protocol of the study. The rats in experimental groups received ethylene glycol and intraperitoneal injection of thymoquinone either from the first day of the study or the 15th day, with either doses of 5 mg/kg or 10 mg/kg. Blood and 24-hour urine samples were collected at baseline and on day 28. Urine oxalate and citrate and serum electrolytes were also measured. On day 29, all rats were decapitated and their kidney specimens were studied. On day 28, urine oxalate concentration significantly decreased in the experimental groups compared to the ethylene glycol group (P < .001). Also, serum calcium levels were significantly higher in the experimental groups (P = .001). Calcium oxalate deposits were smaller in the experimental groups than the ethylene glycol group. The mean number of deposits was lower in these groups, too (P < .001). Treatment with the lower dose of thymoquinone was associated with fewer deposits. Thymoquinone significantly decreased the number and size of calcium oxalate deposits in the renal tubules. The dose and duration of treatment, however, does not have a linear relation with the outcomes. Further studies on thymoquinone as a preventive and therapeutic drug for kidney calculi are suggested.


What Makes Patient Navigation Most Effective: Defining Useful Tasks and Networks.
Given the momentum in adopting patient navigation into cancer care, there is a need to understand the contribution of specific navigator activities to improved clinical outcomes. A mixed-methods study combined direct observations of patient navigators within the Patient Navigation Research Program and outcome data from the trial. We correlated the frequency of navigator tasks with the outcome of rate of diagnostic resolution within 365 days among patients who received the intervention relative to controls. A focused content analysis examined those tasks with the strongest correlations between navigator tasks and patient outcomes. Navigating directly with specific patients (r = 0.679), working with clinical providers to facilitate patient care (r = 0.643), and performing tasks not directly related to their diagnostic evaluation for patients were positively associated with more timely diagnosis (r = 0.714). Using medical records for non-navigation tasks had a negative association (r = -0.643). Content analysis revealed service provision directed at specific patients improved care while systems-focused activities did not.


The role of human papillomavirus type 16/18 genotyping in predicting high-grade cervical/vaginal intraepithelial neoplasm in women with mildly abnormal Papanicolaou results.
The authors compared the predictive value of type 16 and/or 18 human papillomavirus (HPV) versus non-16/18 HPV types for high-grade (grade ≥2) cervical neoplasm/vaginal intraepithelial neoplasm and carcinoma (CIN/VAIN2+) in women with mildly abnormal Papanicolaou (Pap) results (ie, atypical squamous cells of undetermined significance [ASCUS] or low-grade squamous epithelial lesion [LSIL]). The authors retrospectively selected Pap specimens with HPV testing results obtained from 243 women (155 with ASCUS and 88 with LSIL Pap results) in their Department of Pathology. HPV genotyping was performed using the EasyChip HPV blot assay. The Pap specimens with HPV16/18 and non-16/18 HPV types were compared with follow-up biopsy results. Follow-up duration ranged from 1 month to 58 months (mean, 26 months). In total, 58 of 155 specimens (37%) that had ASCUS and 29 of 88 specimens (33%) that had LSIL were positive for HPV16/18. CIN/VAIN2+ biopsies were identified in 43 of 155 women (28%) with ASCUS and in 28 of 88 women (32%) with LSIL. Women with ASCUS and HPV16/18 had a significantly higher rate (43%) of CIN/VAIN2+ than women with ASCUS and non-16/18 HPV types (19%; P = .003; odds ratio, 3.10; 95% confidence interval, 1.48-6.53). There was no statistically significant difference in the rate of CIN/VAIN2+ between women who had LSIL and HPV16/18 (45%) and those who had LSIL and non-16/18 HPV types (29%; P = .16; odds ratio, 1.96; 95% confidence interval, 0.77-4.97). HPV genotyping for HPV16/18 improved risk assessment for women with ASCUS Pap results and may be used to predict the risk of CIN/VAIN2+ to better guide follow-up management.


Comparative aspects of herbicide metabolism.
Herbicides are the economically most important group of pesticides, and such activity occurs in a wide range of chemical structural classes. Studies of the metabolism of herbicides involve a great number of organisms and test systems to provide information of value for assessing the nature of potential residues in human foods of plant and animal origin, metabolism-toxicity relationships in mammals in view of potential human exposure, and in plants in view of biological selectivity, and the environmental fate in soil and aquatic systems. Some of the objectives pursued in an industrial metabolism laboratory and the problems confronted when elaborating its contribution to the safety assessment of a pesticide are briefly described with special reference to metabolism-selectivity relationships in plants and the significance of plant metabolites to animals.


Factors affecting the biotransformation of the pesticide parathion by the isolated perfused mouse liver.
Single-pass perfusion of mouse livers in situ with the phosphorothioate pesticide parathion resulted in formation of paraoxon, p-nitrophenol, p-nitrophenyl sulfate, and p-nitrophenyl-beta-D-glucuronide. At a perfusate bovine serum albumin (BSA) concentration of 4% (fraction of unbound parathion = 0.04), and a flow rate of 3.2 ml/min/liver, the half-life associated with the approach to steady state of parathion was 6.2 min (SD = 0.4), whereas at steady state the extraction ratio of parathion was 0.19 (SD = 0.03). Alterations in perfusate flow rates had no discernable effects on metabolism of parathion. However, lowering the BSA perfusate concentration to 1.0% (fraction of unbound parathion = 0.12) significantly prolonged the half-life for the approach to steady state while increasing the steady state extraction ratio to 0.49 (SD = 0.08). At perfusate BSA concentrations below 1%, steady state conditions with respect to parathion could not be achieved during the 50-min perfusions. These results suggest that binding of parathion to BSA hinders its biotransformation by mouse livers perfused in situ, probably by limiting the availability of free pesticide to metabolic sites. Consequently, its elimination by the liver is insensitive to changes in hepatic blood flow. However, exclusion of BSA from the perfusate resulted in partitioning of all parathion in perfusate into the liver, leading to high concentrations of parathion within liver and preventing biotransformation of this pesticide.


[Higher risk for health care associated infections in hospitalized children with special health needs].
Nosocomial infections (NIs) represent an important public health issue, being associated with high morbidity, mortality and costs. To identify risk factors of NIs in a cohort of hospitalized pediatric patients, with emphasis in children with special health care needs (CSHCN). We studied patients hospitalized in Sótero del Río Pediatric Hospital at Santiago-Chile, between September and December, 2009. Relative Risk (RR) was calculated between CSHCN-3 (children attended by three or more health care professionals) versus non CSHCN-3, and between children with medical complexity (CMC) versus non CMC, using log-binomial models. 920 patients were included. Prevalence of NIs was 12.7%. The most frequent diagnoses were gastrointestinal and respiratory viral infections. Both crude and adjusted by confounders (age and sex), CSHCN-3 and CMC presented significantly higher risk of NIs compared to patients without these conditions (RR 2.91 and RR 2.16, respectively). CSHCN-3 and CMC represent independent conditions associated to a 2-3 fold risk of NIs.


Rupture of the diaphragm due to blunt trauma.
From 1968 to 1980 we have treated 21 patients with a rupture of the diaphragm due to blunt trauma. The diagnosis was generally made in the acute phase. Right-sided rupture in this series accounted for 24% of cases. This figure differs significantly from the 10% incidence mentioned in the literature. In our opinion right-sided rupture, especially in patients with multiple injuries, is often overlooked.


Rabies among African wild dogs (Lycaon pictus) in the Masai Mara, Kenya.
A pack of African wild dogs (Lycaon pictus) ranging to the north of the Masai Mara National Reserve in southwestern Kenya was monitored from 1988 to 1989. During a 6-week period (August 1-September 13, 1989), 21 of 23 members of this pack died. Seven carcasses were retrieved, of which 4 were suitable for necropsy and histopathologic examination. Gross findings varied among individuals and included multiple bite wounds, synovitis, lymphadenopathy, submandibular, cervical, and vocal cord edema, blood in bronchi, bronchioles, stomach, and intestine, and interioventral lung lobe consolidation. Histologic examination of 2 available brain samples revealed nonsuppurative encephalitis with eosinophilic intracytoplasmic inclusions (Negri bodies). An additional brain sample tested positive for rabies via a fluorescent antibody test. Other histologic features included severe suppurative bronchopneumonia, myocarditis, and lymphoid depletion of the lymph nodes, tonsils, and spleen. A 304-base pair (bp) nucleotide sequence from the N gene and a 310-bp sequence from the G gene from rabies isolates of 4 wild dogs indicated that infection was with a rabies variant common among domestic dogs in Kenya and Tanzania.


Stability and infectivity of novel pandemic influenza A (H1N1) virus in blood-derived matrices under different storage conditions.
Influenza A virus has been detected in the blood of some infected individuals, and may pose a safety concern for collection, handling and transport of specimens for epidemiological and public health investigations if infectious virus is present in samples. Furthermore the effect of storage on virus stability and infectivity has not been well studied. We examined the stability of novel pandemic influenza A (H1N1) virus RNA when the virus was stored in phosphate buffered saline (PBS), plasma, or buffy coated blood at either room temperature or 4°C using a sensitive Taqman RT-PCR assay. We also investigated virus infectivity using the EID(50) assay when virus was stored in PBS, plasma, or buffy coats isolated from blood at 4°C. Viral RNA stability was affected by the matrix used for storage. The recovery of viral RNA was highest when virus was stored in PBS with lower amounts being recovered from plasma and buffy coats at either room temperature or 4°C. Incubation time did not appear to be a major factor for viral RNA stability, although there was gradual decline after longer periods post-incubation. Both sample matrix and incubation time affected virus infectivity. The decay in virus infectivity was greatest in PBS followed by buffy coats and plasma. Virus infectivity was abolished in buffy coats at day 20 post-incubation when virus concentrations were low. These data indicate that encapsidated viral RNA was stable overall in all three liquid matrices at room temperature or 4°C although it was most stable in PBS; virus infectivity in buffy coats at 4°C decayed in a time dependent manner while it remained unchanged in plasma. These findings have implications for storage, handling and transport of blood derived samples from influenza patients for epidemiological and laboratory investigations. It should be noted that there is little known about influenza viremia, and whether influenza viruses can be transmitted by blood or blood derived samples.


Evaluating the efficacy of entomopathogenic nematodes for the biological control of crop pests: a nonequilibrium approach.
The efficacy of entomopathogenic nematodes for biological control is assessed using deterministic models. Typically, the examination of such models involves stability analyses to determine the long-term persistence of control. However, in agricultural systems, control is often needed within a single season. Hence, the transient dynamics of the systems were assessed under specific, short-term control scenarios using stage-structured models. Analyses suggest that preemptive application may be the optimum strategy if nematode mortality rates are low; applying before pest invasion can result in greater control than applying afterward. In addition, repeated applications will suppress a pest, providing the application rate exceeds a threshold. However, the period between applications affects control success, so the economic injury level of the crop and the life history of the pest should be evaluated before deciding the strategy. In all scenarios, the most important parameter influencing control is the transmission rate. These findings are applicable to more traditional biological control agents (e.g., microparasites and parasitoids), and we recommend the approach adopted here when considering their practical use. It is concluded that it is essential to consider the specific crop and pest characteristics and the definition of control success before selecting the appropriate control strategy.


Biomarker Supervised G-CSF (Filgrastim) Response in ALS Patients.
Objective: To evaluate safety, tolerability and feasibility of long-term treatment with Granulocyte-colony stimulating factor (G-CSF), a well-known hematopoietic stem cell factor, guided by assessment of mobilized bone marrow derived stem cells and cytokines in the serum of patients with amyotrophic lateral sclerosis (ALS) treated on a named patient basis. Methods: 36 ALS patients were treated with subcutaneous injections of G-CSF on a named patient basis and in an outpatient setting. Drug was dosed by individual application schemes (mean 464 Mio IU/month, range 90-2160 Mio IU/month) over a median of 13.7 months (range from 2.7 to 73.8 months). Safety, tolerability, survival and change in ALSFRS-R were observed. Hematopoietic stem cells were monitored by flow cytometry analysis of circulating CD34+ and CD34+CD38- cells, and peripheral cytokines were assessed by electrochemoluminescence throughout the intervention period. Analysis of immunological and hematological markers was conducted. Results: Long term and individually adapted treatment with G-CSF was well tolerated and safe. G-CSF led to a significant mobilization of hematopoietic stem cells into the peripheral blood. Higher mobilization capacity was associated with prolonged survival. Initial levels of serum cytokines, such as MDC, TNF-beta, IL-7, IL-16, and Tie-2 were significantly associated with survival. Continued application of G-CSF led to persistent alterations in serum cytokines and ongoing measurements revealed the multifaceted effects of G-CSF. Conclusions: G-CSF treatment is feasible and safe for ALS patients. It may exert its beneficial effects through neuroprotective and -regenerative activities, mobilization of hematopoietic stem cells and regulation of pro- and anti-inflammatory cytokines as well as angiogenic factors. These cytokines may serve as prognostic markers when measured at the time of diagnosis. Hematopoietic stem cell numbers and cytokine levels are altered by ongoing G-CSF application and may potentially serve as treatment biomarkers for early monitoring of G-CSF treatment efficacy in ALS in future clinical trials.


Preparation and use of calcium phosphate adsorbed vaccines.
Simple or combined D,T,P and inactivated polio vaccines adsorbed onto calcium phosphate are prepared according to two procedures. Antigens can be dialysed in a sodium phosphate solution and quickly mixed with an equal volume of an equimolar solution of calcium chloride, the pH is adjusted to 6.8-7. Toxoids added to the phosphate solution are in this way entrapped within the 3-dimensional network during the formation of the precipitate. Antigens can also be added to a calcium phosphate gel suspension prepared in advance. Results of animal experiments and of field trials using either combined vaccines or simultaneous immunization with diphtheria, tetanus, pertussis, meningococcal and several viral vaccines: polio, rabies, hepatitis-B, etc., will be presented. Several programs have been studied in developing countries, mainly with the aim at simplifying vaccination campaigns. The efficiency of a two-dose regimen with DT vaccine has been ascertained, this has also been applied to pregnant women. Adsorbed tetanus toxoid was successfully used as a diluent for freeze-dried measles, meningococcal polysaccharide and rabies vaccines. Levels of circulating antibodies and potency of vaccines have been measured by new in vivo and in vitro methods. The choice of detoxified purified toxins is desirable for the preparation of vaccines in order to prevent the occurrence of adverse reactions. Local and generalized reactions have been studied. Adverse Arthus-type reactions have been encountered and related with the presence of high levels of circulating tetanus antibodies.(ABSTRACT TRUNCATED AT 250 WORDS)


Alteration of susceptibility to EDTA, polymyxin B and gentamicin in Pseudomonas aeruginosa by divalent cation regulation of outer membrane protein H1.
Induction of outer membrane protein H1 in Pseudomonas aeruginosa results in decreased susceptibility to aminoglycosides, polymyxin B, and EDTA. We have previously shown that protein H1 can become the major cellular protein in cells grown in low (0.02 mM) Mg2+. The induction of protein H1 was prevented by supplementation of low Mg2+ medium with Mg2+, Ca2+, Mn2+, or Sr2+ (each at 0.5 mM), but not with Zn2+, Ba2+, Sn2+, Al3+ or Na+ (each at 0.5 mM). Only cells grown in the presence of those cations which failed to prevent H1 induction were resistant to the cationic antibiotics, polymyxin B and gentamicin, and to chelators of divalent cations. Cells grown in Ca2+, but not in Mg2+, were susceptible to outer membrane permeabilization by the Ca2+ specific chelator EGTA, whereas both were susceptible to EDTA. In agreement with this, cells grown in Mg2+, Ca2+, Mn2+, or Zn2+ showed enhanced levels of these cations respectively as their major cell envelope-associated cation. When cells were shifted from low to high Mg2+ medium, the time course of the decrease in the levels of protein H1 correlated well with the increase in sensitivity to EDTA and polymyxin B. These results support the hypothesis that protein H1 acts to replace divalent cations at a critical outer membrane site attacked by cationic antibiotics and chelators of divalent cations, and suggest that only a small proportion of the total divalent cation-binding sites in the outer membrane are susceptible to attack by these agents.


Glioma immunology and immunotherapy.
Despite advances in conventional therapy, the prognosis for most glioma patients remains dismal. This has prompted an intensive search for effective treatment alternatives. Immunotherapy, one such alternative, has long been recognized as a potentially potent cancer treatment but has been limited by an inadequate understanding of the immune system. Now, increased insight into immunology is suggesting more rational approaches to immunotherapy. In this article, we explore key aspects of modern immunology and discuss their implications for glioma therapy. A thorough literature review of glioma immunology and immunotherapy was undertaken to inquire into the basic immunology, central nervous system immunology, glioma immunobiology, standard glioma immunotherapy, and recent immunotherapeutic advances in glioma treatment. Although gliomas express tumor-associated antigens and appear potentially sensitive to immune responses, many factors work together to inhibit antiglioma immunity. Not surprisingly, most clinical attempts at glioma immunotherapy have met with little success to date. However, novel immunostimulatory strategies, such as immunogene therapy, directed cytokine delivery, and dendritic cell manipulation, have recently yielded dramatic preclinical results in glioma models. This suggests that glioma-derived immunosuppression can be overcome. Modern molecular biology and immunology techniques have yielded a wealth of new data about glioma immunobiology. Armed with this information, many investigators have proposed novel means to stimulate antiglioma immune responses. Although definitive clinical results remain to be seen, the current renaissance in glioma immunology and immunotherapy shows great promise for the future.


Visual results following infantile cataract surgery.
Aspiration and its modifications have been shown to be a superior operative procedure for neonatal, congenital, infantile, and acquired cataracts. However, the visual results obtained in the infantile cataractous eye, whether the opacity is complete or partial, still remain within the same range as the results of other operative techniques reported over the last 3 decades. Adequate visual results often cannot be obtained because of the associated systemic and/or ocular defects or failure of optical and amblyopia treatment. Perhaps increased efforts should be made to perform surgery at even earlier ages in those patients with complete, or in some cases, partial, cataracts in which surgery is contemplated. In children with partial cataracts who are able to function well, the lens should be allowed to remain in place and conservative measures of visual preservation should be undertaken for as long as possible. This allows the eye more time to develop normal visual functions with an intact lens and accommodative mechanisms. It should be stressed again that the sole success or failure of surgery for infantile cataracts cannot always be the visual acuity. The real measure is the child's ability to function successfully in his environment for the remainder of his life.


Foggy days and dry nights determine crown-level water balance in a seasonal tropical Montane cloud forest.
The ecophysiology of tropical montane cloud forest (TMCF) trees is influenced by crown-level microclimate factors including regular mist/fog water inputs, and large variations in evaporative demand, which in turn can significantly impact water balance. We investigated the effect of such microclimatic factors on canopy ecophysiology and branch-level water balance in the dry season of a seasonal TMCF in Veracruz, Mexico, by quantifying both water inputs (via foliar uptake, FU) and outputs (day- and night-time transpiration, NT). Measurements of sap flow, stomatal conductance, leaf water potential and pressure-volume relations were obtained in Quercus lanceifolia, a canopy-dominant tree species. Our results indicate that FU occurred 34% of the time and led to the recovery of 9% (24 ± 9.1 L) of all the dry-season water transpired from individual branches. Capacity for FU was independently verified for seven additional common tree species. NT accounted for approximately 17% (46 L) of dry-season water loss. There was a strong correlation between FU and the duration of leaf wetness events (fog and/or rain), as well as between NT and the night-time vapour pressure deficit. Our results show the clear importance of fog and NT for the canopy water relations of Q. lanceifolia.


Cutting edge: the NLRP3 inflammasome links complement-mediated inflammation and IL-1β release.
The complement system is a potent component of the innate immune response, promoting inflammation and orchestrating defense against pathogens. However, dysregulation of complement is critical to several autoimmune and inflammatory syndromes. Elevated expression of the proinflammatory cytokine IL-1β is often linked to such diseases. In this study, we reveal the mechanistic link between complement and IL-1β secretion using murine dendritic cells. IL-1β secretion occurs following intracellular caspase-1 activation by inflammasomes. We show that complement elicits secretion of both IL-1β and IL-18 in vitro and in vivo via the NLRP3 inflammasome. This effect depends on the inflammasome components NLRP3 and ASC, as well as caspase-1 activity. Interestingly, sublethal complement membrane attack complex formation, but not the anaphylatoxins C3a and C5a, activated the NLRP3 inflammasome in vivo. These findings provide insight into the molecular processes underlying complement-mediated inflammation and highlight the possibility of targeting IL-1β to control complement-induced disease and pathological inflammation.


Differences in cancer incidence among Indians in Alaska and New Mexico and U.S. Whites, 1993-2002.
Cancer incidence for American Indians and Alaska Natives is typically reported as a single rate for all U.S. indigenous populations combined. Previously reported combined rates suggest that American Indians and Alaska Natives have lower cancer incidence rates compared with the U.S. population. Alaska Native people comprise three major ethnic groups: Eskimo, Indian, and Aleut people. We examined cancer incidence from only Alaska Indians and compared incidence rates with an American Indian population living in New Mexico. These data indicate striking differences in cancer patterns between two American Indian populations. Cancer data for the years 1993 to 2002 for American Indians of New Mexico and U.S. Whites are from the National Cancer Institute Surveillance, Epidemiology, and End Results Program Public-use data set. Data for Alaska Indians are from the Alaska Native Tumor Registry, which is also a Surveillance Epidemiology and End Results Program participant. Overall, cancer incidence rates for all sites combined in New Mexico Indian men and women were lower than U.S. White rates, whereas Alaska Indian men and women exceeded U.S. rates. In comparing Alaska and New Mexico Indians, we observed a 2.5-fold higher incidence of cancer among Alaska Indians. The largest differences between the two Indian populations were noted primarily in cancers associated with tobacco use, including cancers of the oral cavity/pharynx, esophagus (only in men), colon and rectum, pancreas, larynx (men), lung, prostate, and urinary bladder (men). Lung cancer rates in Alaska Indian men and women were 7 and 10 times those of New Mexico Indian men and women.


Bolivian ectoparasites: a new species of laelapine mite (Acari: Parasitiformes, Laelapidae) from the rodent Neacomys spinosus.
Laelaps neacomydis n. sp. is described from the pelage of the rodent Neacomys spinosus collected in Bolivia. Collection records from northern Brazil, southeastern Peru, and central Bolivia indicate its wide distribution across Amazonas associated with rodents of the genus Neacomys. The formal taxonomic description and illustrations are derived from the adults (female and male). Two larger species of laelapine mites, Gigantolaelaps intermedius and L. boultoni, infest the same hosts at the same localities.


Transformation of prostaglandin D2 to 9 alpha, 11 beta-(15S)-trihydroxyprosta-(5Z,13E)-dien-1-oic acid (9 alpha, 11 beta-prostaglandin F2): a unique biologically active prostaglandin produced enzymatically in vivo in humans.
In vitro studies examining the metabolic transformation of prostaglandin D2 (PGD2) by human liver were conducted. PGD2 was found to be converted by a NADPH-dependent enzyme in the 100,000 X g supernatant of human liver exclusively to a single more polar compound that had a mass spectrum essentially the same as that of prostaglandin F2 alpha (PGF2 alpha). However, this compound could be chromatographically separated from PGF2 alpha and failed to form a butylboronate derivative. The structure of this compound was established as 9 alpha, 11 beta-(15S)-trihydroxyprosta-(5Z, 13E)-dien-1-oic acid (9 alpha, 11 beta-PGF2) by comparison of its chromatographic and mass spectral characteristics with authentic 9 alpha, 11 beta-PGF2. This compound was found to be biologically active by demonstrating increases in blood pressure in rats in a dose-related fashion following intravenous administration. By using a mass spectrometric assay, levels of this compound in plasma and urine from a normal volunteer were 6 pg/ml and 982 ng/24 hr, respectively. In a patient with systemic mastocytosis associated with overproduction of PGD2, urinary excretion of 9 alpha, 11 beta-PGF 2 was 6634 ng/24 hr and a circulating plasma level as high as 490 ng/ml was found during a severe episode of systemic mast cell activation. 9 alpha, 11 beta-PGF2 is structurally a unique prostaglandin, is enzymatically formed, is produced in vivo in humans, and is biologically active.


Protein kinase Cδ mediates MCP-1 mRNA stabilization in vascular smooth muscle cells.
Monocyte chemoattractant protein-1 (MCP-1) is an inflammatory chemokine that promotes atherosclerosis and is a mediator of the response to arterial injury. We previously demonstrated that platelet-derived growth factor (PDGF) and angiotensin II (Ang) induce the accumulation of MCP-1 mRNA in vascular smooth muscle cells mainly by increasing mRNA stability. In the present study, we have examined the signaling pathways involved in this stabilization of MCP-1 mRNA. The effect of PDGF (BB isoform) and Ang on MCP-1 mRNA stability was mediated by the PDGF β and angiotensin II receptor AT1R, respectively, and did not involve transactivation between the two receptors. The effect of PDGF-BB was blocked by inhibitors of protein kinase C (PKC), but not by inhibitors of phosphoinositol 3-kinase (PI3K), Src, or NADPH oxidase (NADPHox). In contrast, the effect of Ang was blocked by inhibitors of Src, and PKC, but not by inhibitors of PI3 K, or NADPHox. The effect of PDGF BB on MCP-1 mRNA stability was blocked by siRNA directed against PKCδ and protein kinase D (PKD), whereas the effect of Ang was blocked only by siRNA directed against PKCδ. These results suggest that the enhancement of MCP-1 mRNA stability by PDGF-BB and Ang are mediated by distinct "proximal" signaling pathways that converge on activation of PKCδ. This study identifies a novel role for PKCδ in mediating mRNA stability in smooth muscle cells.


Obesity is associated with an increase in pharmaceutical expenses among university employees.
To examine costs associated with obesity in an employee population and factors associated with increased costs. We used data from the Physical Activity and Life Style (PALS) study, a randomized prospective design evaluating three interventions to increase physical activity among physically inactive nonfaculty university employees (n = 454). The primary exposure variable, obesity (measured by body mass index), was obtained from the in-person baseline survey. Covariates were obtained from the baseline survey and included demographic characteristics and health status. Data from the baseline survey was linked with administrative data to determine pharmaceutical, inpatient, outpatient, and total health care costs for three years. Average monthly expenditures for obese and nonobese individuals were compared using t-tests and a two-part multivariate regression model adjusted for demographic and socioeconomic characteristics and health behaviors. Although in-patient and outpatient expenses were not associated with obesity, pharmaceutical expenditures were $408 or 87.2% higher per year ($468 versus $876) for obese individuals than for nonobese individuals, which reflected poorer health behaviors and health status of obese adults. Awareness of the costs associated with obesity among employees can stimulate employers to make the investment in providing employer-sponsored wellness and health improvement programs to address obesity.


Bead study: a novel method to measure gastrointestinal transit in mice.
Intestinal transit assessment in mice using existing methods requires long recording periods or euthanization of animals to localize a tracer. We have developed a novel in vivo method to assess gastrointestinal (GI) transit in mice based on a clinically used 'shapes study'. Mice (n = 70) were gavaged with 5 steel beads and barium 3 h before, with another dose of barium gavaged 10 min before imaging. Mice were fluoroscoped for 20-60 s, and then most of them were euthanized and the GI tract removed to confirm the localization of the beads fluoroscopically. The in vivo and postmortem recordings were analyzed and each bead was scored depending on its location; a total score was calculated by adding individual bead scores. Total scores obtained from the two methods were compared. A group of mice (n = 10) were examined on three occasions, before and after treatment with loperamide or prucalopride. The stomach and cecum were consistently outlined by barium, serving as reference landmarks. There was an excellent overall correlation between in vivo and postmortem transit scores (r = 0.93). Analysis of scores for individual gut segments revealed high agreement for stomach, cecum, and expelled beads, and moderate agreement for the small bowel and colon. Gastrointestinal transit scores were decreased by loperamide and increased by prucalopride compared with baseline. Metallic beads are reliably localized by videofluoroscopy in vivo within the GI tract. This novel imaging method enables repetitive measurements of GI transit in vivo and detects changes induced by motility-modifying agents.


Derivation, characterization, and stable transfection of induced pluripotent stem cells from Fischer344 rats.
The rat represents an important animal model that, in many respects, is superior to the mouse for dissecting behavioral, cardiovascular and other physiological pathologies relevant to humans. Derivation of induced pluripotent stem cells from rats (riPS) opens the opportunity for gene targeting in specific rat strains, as well as for the development of new protocols for the treatment of different degenerative diseases. Here, we report an improved lentivirus-based hit-and-run riPS derivation protocol that makes use of small inhibitors of MEK and GSK3. We demonstrate that the excision of proviruses does not affect either the karyotype or the differentiation ability of these cells. We show that the established riPS cells are readily amenable to genetic manipulations such as stable electroporation. Finally, we propose a genetic tool for an improvement of riPS cell quality in culture. These data may prompt iPS cell-based gene targeting in rat as well as the development of iPS cell-based therapies using disease models established in this species.


In vitro marrow purging in chronic myelogenous leukemia: effect of mafosfamide and recombinant granulocyte--macrophage colony-stimulating factor.
Clinical and experimental evidence revealing Ph1-negative hematopoietic stem cells in the majority of chronic myelogenous leukemia (CML) patients, suggests that autologous bone marrow transplantation (ABMT) may represent a therapeutic approach for these patients. It was the aim of the present study to evaluate the efficacy of the cyclophosphamide derivative mafosfamide as a marrow purging agent in a group (n = 15) of CML patients. Chemical purging was followed by a short-term liquid culture phase supplemented with recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF). Mafosfamide (100 micrograms/ml) incubation induced a marked inhibition of progenitor cell growth, the percentages of surviving CFU-GEMM, BFU-E, and CFU-GM being 3.4, 5.4, and 4.9, respectively. At the cytogenetic level, the purging procedure failed to show any modulating effect on Ph1-negative clones in 9/15 cases. In contrast, 6/15 cases showed a significant increase in the mean (+/- SD) percentage of Ph1-negative metaphases in response to rGM-CSF (46 +/- 26, p less than or equal to 0.05), mafosfamide incubation (53 +/- 12, p less than or equal to 0.01), and the combination of mafosfamide incubation plus rGM-CSF (63 +/- 29, p less than or equal to 0.025). Immunological analysis revealed that mafosfamide incubation induced a significant enrichment of MY10 (28 +/- 9, 0.05) B73.1-positve cells (25 +/- 9, p less than or equal to 0.05). Four mafosfamide-responsive patients with CML in second chronic phase have been autografted with mafosfamide purged marrow. In all patients a Ph1-negative phase lasting 5-14 months was observed. In conclusion, it appears that (a) in a subgroup of CML patients mafosfamide purging is effective in reducing the size of the malignant clone and might induce through its cytotoxic and immune actions a modification of the balance between leukemic and normal clones, and (b) this experimental approach may be used as a screening test to select patients to undergo marrow harvest and ABMT with mafosfamide purged marrow.


Perceived learning needs of Yemeni patients after coronary artery bypass graft surgery.
To explore the information needs of Yemeni patients who have undergone coronary artery bypass graft surgery before hospital discharges and to examine the differences in the patients' learning needs according to age, gender, level of education and working condition. Postcoronary artery bypass graft (CABG) patients need more information before their discharge from the hospital. This need of information must be assessed and provided for them before their discharge from hospital to help them with adequate care and recovery at home. Design. A descriptive, correlational design. Data were collected from 120 CABG patients before their discharge from Al- Thawra Hospital, Sana'a, Yemen, by using modified Cardiac Patients Learning Needs Inventory (CPLNI) instrument. The Statistical Package for the Social Sciences was used to analyse the collected data. The information about chest and leg wound care, medication information, other pertinent information and complications were indicated to be the most valued by CABG patients prior to discharge. There was a statistically significant difference between patients' total learning needs according to their socio-demographic characteristics. Male patients needed more information than female patients (p=0.004). Younger (p=0.021) and middle-aged (p=0.032) patients needed more information than older ones. Highly educated (p=0.000) and school-level (p=0.002) patients needed more information than those in the uneducated group. Working patients needed more information than non-working ones (p=0.000). This study suggests that CABG patients have high information needs within 24-48 hours before hospital discharge. In addition, the results of this study showed that there are statistically significant differences in patients' learning needs according to their demographic characteristics. Assessment of CABG patients' information needs before their discharge from hospital helps in developing effective educational programme that will help these patients in their recovery at home.


Granulocyte-macrophage colony-stimulating factor and other cytokines regulate surface expression of the leukocyte adhesion molecule-1 on human neutrophils, monocytes, and their precursors.
There is increasing evidence that cytokines such as granulocyte-macrophage (GM)-CSF can profoundly affect the adhesion, aggregation, and mobility of neutrophils both in vitro and in vivo. However, the mechanisms whereby these factors might alter the adhesive properties of neutrophils are incompletely understood. A new family of cellular adhesion molecules has recently been identified by cDNA cloning. The members of this family include human leukocyte adhesion molecule-1 (LAM-1), the human endothelial-leukocyte adhesion molecule, and the mouse leukocyte homing receptor for high endothelial venules, MEL-14. LAM-1 is the human homologue of murine MEL-14, and is believed to mediate binding of leukocytes to human high endothelial venules. LAM-1 can be identified by mAb TQ-1, Leu 8, or anti-LAM1.1. The expression and regulation of LAM-1 on granulocytes, monocytes, and their precursors was investigated using flow cytometry and the anti-LAM-1.1 mAb. Neutrophils, eosinophils, monocytes, marrow myeloid cells, granulocyte/macrophage colony-forming unit, and burst-forming unit for erythroid cells were LAM-1+ by flow microfluorimetry. The regulation of LAM-1 expression was tested by treating various cell populations with cytokines or other stimuli for 0-90 min. Exposure of neutrophils, monocytes, and marrow myeloid cells to GM-CSF induced rapid and complete loss of LAM-1 from the cell surface, but had no effect on LAM-1 expression by lymphocytes. The loss of LAM-1 was temporally correlated with up-regulation of CD11b (Mo1), an adhesion molecule involved in neutrophil aggregation. Several other factors known to activate neutrophils also caused down-regulation of LAM-1 and up-regulation of CD11b, including TNF, FMLP, and leukotriene B4. Interestingly, granulocyte-CSF and IFN-gamma had minimal effects on neutrophil LAM-1 expression. Similar results were observed on monocytes and myeloid precursor cells. Thus, exposure of neutrophils to GM-CSF results in a profound change in surface expression of adhesion molecules, with coordinated up-regulation of CD11b and down-regulation of LAM-1. These changes in adhesion proteins are likely to alter aggregation and mobility of both mature myeloid cells and their precursors in patients receiving certain types of cytokine therapy.


Inheritance of a mutant histocompatibility gene and a new mammary tumor virus genome in the B6.KH-84 mouse strain.
The potential association between integration or deletion of mouse mammary tumor virus (MMTV) retroviral sequences and the appearance of non-H-2 histocompatibility (H) antigen mutations was investigated. Genomic blots from inbred strains carrying 22 loss, gain-loss, and gain mutations on the BALB/c and C57BL/6 backgrounds were hybridized with probes homologous to the long terminal repeat (LTR) and envelope (env) regions of MMTV. Twenty-one mutants were identical in restriction patterns to the respective background strains with all tested restriction enzymes and both probes. However, genomic blots of one gain mutant, B6.C-KH-84, exhibited restriction fragments which were not exhibited by either of the parental strains, C57BL/6 or BALB/c. An additional 5.5 kb Eco RI fragment was observed with the env probe and additional 9.2 kb and 5.5 kb fragments were observed with the LTR probe. These observations were substantiated by hybridization of these two probes with genomic blots generated with additional restriction enzymes. Assuming that the new provirus contains a single, internal Eco RI site as has been observed for other MMTV proviral sequences, it is presumed that the new provirus includes both 5' and 3' LTRs in addition to the env region. Based on the unique sizes of the observed restriction fragments relative to other identified MMTV proviral sequences, this provirus has been designated Mtv-22. The potential role of Mtv-22 in the genesis of the gained histocompatibility antigen in B6.C-KH-84 is discussed.


[Modern approaches to beta-interferon application in therapy of multiple sclerosis].
Recently a disease-modifying treatment of multiple sclerosis (MS) became possible. The first choice in this kind of treatment is the use of beta-interferons. The longest history has the studies of beta-interferon-1b, which has a beneficial effect on the course of MS reducing the annual exacerbation rate and slowing the disability progression. This kind of treatment is very expensive and has only a moderate influence on the disease course, only slowing its progression, but still are widely used and causes attention all over the world. The most important question is the appropriate selection of MS patients for this kind of treatment. Numerous regional neurological community has elaborated special recommendations for the use of this treatment. These recommendations are usually base on the data of clinical trials, own experience and current financial situation (to cover the high cost of this treatment). Analyses of the last data on the mechanisms of beta-interferons treatment showed several specific and nonspecific effects. In recently published studies beta-interferon-1b was shown to slow the progression of confirmed disability in secondary progressive MS and delay the switch from relapsing/remitting to secondary progressive MS. Still the most beneficial clinical effect is seen in relapsing/remitting cases. The problem of neutralizing antibodies (NAT) and treatment of possible side effects seem now not to be so principle. Recommendations for the selection of patients with MS for treatment with beta-interferon were proposed.


Epistatic genetic effects among Alzheimer's candidate genes.
Novel risk variants for late-onset Alzheimer's disease (AD) have been identified and replicated in genome-wide association studies. Recent work has begun to address the relationship between these risk variants and biomarkers of AD, though results have been mixed. The aim of the current study was to characterize single marker and epistatic genetic effects between the top candidate Single Nucleotide Polymorphisms (SNPs) in relation to amyloid deposition. We used a combined dataset across ADNI-1 and ADNI-2, and looked within each dataset separately to validate identified genetic effects. Amyloid was quantified using data acquired by Positron Emission Tomography (PET) with (18)F-AV-45. Two SNP-SNP interactions reached significance when correcting for multiple comparisons, BIN1 (rs7561528, rs744373) x PICALM (rs7851179). Carrying the minor allele in BIN1 was related to higher levels of amyloid deposition, however only in non-carriers of the protective PICALM minor allele. Our results support previous research suggesting these candidate SNPs do not show single marker associations with amyloid pathology. However, we provide evidence for a novel interaction between PICALM and BIN1 in relation to amyloid deposition. Risk related to the BIN1 minor allele appears to be mitigated in the presence of the PICALM protective variant. In that way, variance in amyloid plaque burden can be better classified within the context of a complex genetic background. Efforts to model cumulative risk for AD should explicitly account for this epistatic effect, and future studies should explicitly test for such effects whenever statistically feasible.


Role of C-type lectins in mycobacterial infections.
Worldwide clinical cases due to multi drug- and extensively drug-resistant strains of Mycobacterium tuberculosis (M.tb) are increasing making the need for new therapies more critical than ever. A major obstacle for designing new drugs to treat mycobacterial infections is our limited knowledge of the interface between the bacillus (especially M.tb) and its host. The pulmonary innate immune system plays a key role in the recognition of microbes entering via the respiratory route. Although the specificity of this system is broad and based on the recognition of pathogen-associated molecular patterns (PAMPs), it is uniquely regulated to limit inflammation and thereby prevent damage to the gas-exchanging alveoli. Pulmonary surfactant proteins A and D (SP-A and SP-D) are collagenous, soluble, C-type (Ca(2+)-dependent) lectins (named collectins) of the lung innate immune system that are secreted into the alveoli by resident type II alveolar epithelial cells and distal bronchiolar Clara cells. The related collectin in serum, mannose-binding lectin/protein (MBL or MBP), provides first-line defense against several microbes. Phagocytes represent the first cellular defense in the alveoli and their surface is rich in C-type lectin pattern recognition receptors (PRRs), including the mannose receptor (MR), dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN) and DC-associated C-type lectin-1 (Dectin-1). This review will discuss the important roles of the cell-associated C-type lectin PRRs and soluble collectins in the innate immune response to mycobacterial infections, and will present the current state of knowledge regarding the potential uses of these C-type lectins in therapy against infections, focusing on M.tb.


Parental smoking and alcohol consumption and risk of neuroblastoma.
Previous studies and animal evidence have suggested a relationship between parental tobacco or alcohol use and the risk of some childhood cancers, including neuroblastoma. A case-control study was conducted to investigate the relationship between parental tobacco smoking, alcohol consumption, and risk of neuroblastoma. Cases were children diagnosed with neuroblastoma over the period 1992-1994 at Children's Cancer Group and Pediatric Oncology Group institutions throughout the United States and Canada. One matched control was selected using random-digit dialing. Information on parental smoking and drinking history was obtained from 504 case and 504 control parents by telephone interview. Overall, there was no consistent pattern of association with parental smoking and alcohol consumption. For example, both maternal smoking and drinking during the period from 1 month before pregnancy through breastfeeding had adjusted odds ratios (ORs) of 1.1 [95% confidence interval (CI), 0.8-1.4]. There was no association with paternal smoking (OR, 1.2; 95% CI, 0.8-1.6) or paternal drinking 1 month before conception (OR, 1.0; 95% CI, 0.7-1.4). There was no consistent increase in risk by the amount of smoking or drinking during any time period relative to pregnancy. There was no suggestion of an increased risk when only one parent smoked. Smoking or drinking among both parents did not jointly increase the risk of neuroblastoma in their offspring. The child's age at diagnosis, stage, or MYCN oncogene amplification status did not materially alter the OR estimates. It is concluded that the results from this study do not indicate any evidence for a relationship between neuroblastoma and parental tobacco or alcohol use.


Evaluation of an online orientation program for new healthcare employees.
Online learning is one of the newest efforts of mainstream healthcare education. It has been cited as providing both an efficient and effective alternative learning medium. This study explored the acceptance of an online orientation program by new healthcare employees and identified several critical factors affecting online learning outcomes. The study applied triangulation methods including surveys, interviews, and test scores. Data were captured from 154 questionnaires (with a response rate of 78.2%) with related test scores and 13 participant interviews. The results showed that new healthcare employees were positive toward the computer-based learning program. "Perceived ease of use" and "perceived usefulness" were the major variables predicting technology acceptance, whereas "learner satisfaction" and "continued intention to use" were the major indices for the learning outcomes. Test scores, however, did not reveal any statistical impact. Additionally, human-computer interaction, instant feedback, and program content were commented on as factors affecting learning outcomes. Therefore, online learning is suitable for new healthcare employees, both in the orientation stage and as a follow-up learning method. The results of this study can help support the development of educational technology online programs.


Recent advances in electrochemical aptasensors and genosensors for the detection of pathogens.
In recent years, pathogenic microorganisms have caused significant mortality rates and antibiotic resistance and triggered exorbitant healthcare costs. These pathogens often have high transmission rates within human populations. Rapid diagnosis is crucial in controlling and reducing the spread of pathogenic infections. The diagnostic methods currently used against individuals infected with these pathogens include relying on outward symptoms, immunological-based and, some biomolecular ones, which mainly have limitations such as diagnostic errors, time-consuming processes, and high-cost platforms. Electrochemical aptasensors and genosensors have emerged as promising diagnostic tools for rapid, accurate, and cost-effective pathogen detection. These bio-electrochemical platforms have been optimized for diagnostic purposes by incorporating advanced materials (mainly nanomaterials), biomolecular technologies, and innovative designs. This review classifies electrochemical aptasensors and genosensors developed between 2021 and 2023 based on their use of different nanomaterials, such as gold-based, carbon-based, and others that employed other innovative assemblies without the use of nanomaterials. Inspecting the diagnostic features of various sensing platforms against pathogenic analytes can identify research gaps and open new avenues for exploration.


Magnetic techniques for the isolation and purification of proteins and peptides.
Isolation and separation of specific molecules is used in almost all areas of biosciences and biotechnology. Diverse procedures can be used to achieve this goal. Recently, increased attention has been paid to the development and application of magnetic separation techniques, which employ small magnetic particles. The purpose of this review paper is to summarize various methodologies, strategies and materials which can be used for the isolation and purification of target proteins and peptides with the help of magnetic field. An extensive list of realised purification procedures documents the efficiency of magnetic separation techniques.


Loss of miR-192-5p initiates a hyperglycolysis and stemness positive feedback in hepatocellular carcinoma.
Emerging studies revealed that cancer stem cells (CSCs) possessed peculiar metabolic properties, which however remained largely unknown in hepatocellular carcinoma (HCC). Genetic silencing of liver-abundant miR-192-5p was a key feature for multiple groups of CSC-positive HCCs. We thus aimed to investigate essential metabolic features of hepatic CSCs via using HCCs with miR-192-5p silencing as a model. Datasets from two independent HCC cohorts were used. Data integration analyses of miR-192-5p with metabolome and mRNA transcriptome data in HCC Cohort 1 were performed to investigate miR-192-5p related metabolic features, which was further validated in Cohort 2. Cellular and molecular assays were performed to examine whether and how miR-192-5p regulated the identified metabolic features. Co-culture systems consisting of HCC cells and LX2 (human hepatic stellate cell line) or THP1 (human monocyte cell line) were established to explore effects of the identified metabolic properties on stemness features of HCC cells via interacting with co-cultured non-tumor cells. High levels of glycolysis-related metabolites and genes were present in HCCs with low miR-192-5p and CSC-positive HCCs in two independent HCC cohorts. miR-192-5p knockout cells displayed CSC features and miR-192-5p loss led to an enhanced glycolytic phenotype via upregulating three bona fide targets, GLUT1 and PFKFB3 (two glycolytic enzymes) and c-Myc (regulating glycolytic genes' expression). Meanwhile, c-Myc suppressed miR-192-5p transcription, ensuring a low-miR-192-5p/high-c-Myc loop to maintain hyperglycolysis. Moreover, over-produced lactic acid from hyperglycolytic HCC cells stimulated the ERK phosphorylation of co-cultured LX2 and THP1 non-tumor cells partially via NDRG3 and MCT1, which in turn promoted cell malignancy and stemness of HCC cells. Consistently, HCC patients with low level of miR-192-5p in their tumor tissues and high level of NDRG3 or MCT1 in their non-tumor tissues had the shortest overall survival. In CSC-positive HCCs, miR-192-5p loss enhanced glycolysis and over produced lactate might further increase HCC malignant features via interacting with environmental non-tumor cells.


How to select aspirant laparoscopic surgical trainees: establishing concurrent validity comparing Xitact LS500 index performance scores with standardized psychomotor aptitude test battery scores.
Although a controversial topic in medical education, the selection of aspirant surgical trainees is a subject that needs to be addressed. In the view of preventing surgical trainee drop-outs and of appropriate allocation of limited resources, it is an issue critical to the profession. Traditional methods of selection are often subjective, and do not seem to correlate with skill needed for surgery. Standardized neuropsychometric test batteries may be useful in helping to select aspirant laparoscopic surgeons. Our study attempts to link surgical novices' psychometric ability test battery data with actual performance outcome on an objective, validated, and reproducible surgical laparoscopic task using virtual-reality simulation. Thirty-three novices with no laparoscopic surgical experience participated. Each participant performed the Xitact LS500 Virtual Reality cholecystectomy clip-and-cut module 30 times. Individual learning curves were computed and patterns were assessed. Participants were examined using the aptitude test battery including the Abstract Reasoning test, the Space Relations test, the Gibson Spiral Maze test, and the Crawford Small Parts Dexterity tester. Over 900 virtual-reality simulation tasks were generated and assessed. Of the participants, 93.3% were able to complete the virtual-reality simulation 30 times and all elements of our aptitude test battery. The abstract reasoning test is the only test correlating significantly to Xitact test outcome. This test is highly correlated to the space relations test. Both tests have discriminative power, comparing groups of performers. The present study addresses the concurrent validity in aptitude testing, comparing scores of surgical novices on the Xitact LS500 laparoscopic cholecystectomy virtual-reality simulation with performance scores on a battery of standardized psychometric aptitude tests. The abstract reasoning and the space-relation test have predictive and selective value, identifying individuals who have good laparoscopic surgical virtual-reality performance. Aspiring laparoscopic surgeons who score below 25 on either test, that is, an expected 36%, would have to be further assessed using Xitact surgical task performance. The group of participants scoring above 35 on the Abstract Reasoning test and above 45 on the Space Relations test, that is, an expected 18% of the population, is unlikely to mal-perform on Xitact. The other 46% could very well benefit from Xitact simulation and assessment when the opportunity is present.


Active compound of Zingiber cassumunar Roxb. down-regulates the expression of genes involved in joint erosion in a human synovial fibroblast cell line.
Rheumatoid arthritis (RA) is a chronic inflammatory disease of the synovium. It is involved in up-regulation of pro-inflammatory cytokines and matrix metalloproteinases (MMPs), resulting in joint inflammation and erosion. Zingiber cassumunar Roxb. has long been used to reduce joint pain and inflammation. This study aimed to investigate the inhibitory activities of an active compound of Z. cassumunar, (E)-4-(3',4'-dimethoxyphenyl)but-3-en-1-ol (compound D), against cytokine-induced up-regulation of catabolic genes involved in cartilage degradation in RA. Synovial fibroblast cell line, SW982, was cultured in media containing interleukin-1β (IL-1β), in the presence or absence of compound D at the concentration range of 1 to 100 µM. After 24 hours, the cells were analyzed for the expressions of MMPs, IL-1β and interleukin-1β-converting enzyme (ICE) by RT-PCR. MMPs activities in the culture media were analyzed by zymographic techniques. Dexamethasone was used as the positive control. It was found that compound D at the concentration of 10 - 100 µM significantly decreased the mRNA expressions of MMP-1, -2, -3, and -13 which was induced by IL-1β (P<0.05) concomitantly with a decrease in activities of these MMPs in the culture media. An increase in the mRNA expression of IL-1β and ICE was also suppressed by compound D. The results suggest that the potent activities of this compound may be involved in the reduction of IL-1β protein synthesis in both pro-form and active form which played an important role in up-regulation of MMPs. This study first revealed the chondroprotective activity of Z. cassumunar in the transcriptional level by suppressing cytokine-induced catabolic genes which caused cartilage erosion in RA.


Pancreatic pseudocysts following acute pancreatitis.
Pancreatic pseudocysts (PP) following acute pancreatitis have traditionally been approached by observation to allow cyst maturation. However, recent evidence suggests a selective approach toward management is indicated. We retrospectively reviewed the presentation, operative management, and outcome of patients developing PP following acute pancreatitis since 1988. PP related to chronic pancreatitis were excluded. Twenty-seven patients were identified, 17 with giant PP (> 10 cm) and 10 with PP less than 10 cm. Groups were not different with respect to age and etiology of pancreatitis, predominantly biliary. Patients with giant PP had a significantly greater number of Ranson criteria at presentation and underwent drainage procedures earlier following their initial presentation. With respect to giant PP, 7 patients underwent internal drainage all via cystogastrostomy with 5 complications. Nine of 10 patients underwent urgent operation via external drainage. There were 3 pancreatic fistulas in this group. The morbidity and mortality rates for giant PP were 65% and 18% respectively. With respect to smaller PP, 8 underwent internal drainage with 1 death (mortality rate 10%). External drainage was performed in 2 patients with 1 pancreatic fistula (morbidity 10%). Patients with PP and a high Ranson score following acute pancreatitis are at significant risk for giant PP formation. Expectant management of giant PP is associated with higher morbidity and mortality than small PP suggesting that earlier external drainage, before clinical deterioration, may be beneficial. To be accurate, comparisons of outcomes for various treatment modalities must take into consideration PP size.


Three nuclear protein-coding genes corroborate a recent phylogenomic model of the Branchiopoda (Crustacea) and provide estimates of the divergence times of the major branchiopodan taxa.
The class Branchiopoda (Crustacea) shows great diversity in morphology and lifestyle among its constituent higher-level taxa: Anostraca, Notostraca, Laevicaudata, Spinicaudata, Cyclestherida and Cladocera. The phylogenetic relationships among these taxa have long been controversial. We sequenced three orthologous nuclear genes that encode the catalytic subunit of DNA polymerase delta and the largest and second-largest subunits of RNA polymerase II in the expectation that the amino acid sequences encoded by these genes might be effective in clarifying branchiopod phylogeny and estimating the times of divergence of the major branchiopodan taxa. The results of phylogenetic analyses based on these amino acid sequences support the monophyly of Branchiopoda and provide strong molecular evidence in support of the following phylogenetic relationships: (Anostraca, (Notostraca, (Laevicaudata, (Spinicaudata, (Cyclestherida, Cladocera))))). Within Cladocera, comparison of the nucleotide sequences of these same genes shows Ctenopoda to be the sister group of Haplopoda + Anomopoda. Three statistical tests based on the present amino acid sequence data-the approximately unbiased test, Kishino-Hasegawa test and weighted Shimodaira-Hasegawa test-tend to refute most of the previous molecular phylogenetic studies on Branchiopoda, which have placed Notostraca differently than here; however, our results corroborate those of one recent phylogenomic study, thus confirming the effectiveness of these three genes to investigate relationships among branchiopod higher taxa. Divergence time estimates calibrated on the basis of fossil evidence suggest that the first divergence of extant branchiopods occurred about 534 Ma during the early Cambrian period and that diversification within the extant branchiopod lineages started in or after the late Permian.


Differential expression of miR-31 between inflammatory bowel disease and microscopic colitis.
Idiopathic inflammatory bowel disease (IBD) and microscopic colitis (MC) are distinct entities. However, patients with intermittent episodes of IBD and MC that are encountered in a clinical setting puzzle clinicians and pathologists. This study examined whether microRNA assisted in the classification of IBD and MC. Small RNA was extracted from formalin-fixed, paraffin-embedded (FFPE) colon tissue and qRT-PCR was performed from cohorts of normal control (n=38), ulcerative colitis (n=36), Crohns disease (n=26), collagenous colitis (n=36), lymphocytic colitis (n=30), and patients with intermittent features of IBD and MC (n=6). Differential expression of miR-31 distinguished IBD (ulcerative colitis and Crohns disease) from MC (collagenous colitis and lymphocytic colitis), confirming the specificity of miR-31 expression in IBD (P=0.00001). In addition, expression of miR-31 was increased in collagenous colitis compared to that of lymphocytic colitis (P=0.010). Among 6 patients with alternating episodes of IBD and MC, one patient had matching miR-31 expression in different phases (lymphocytic colitis to ulcerative colitis, and then back to collagenous colitis). The other 5 patients had MC-like expression patterns in both MC and IBD episodes. In summary, IBD and MC have distinct miR-31 expression pattern. Therefore, miR-31 might be used as a biomarker to distinguish between IBD and MC in FFPE colonic tissue. In addition, miR-31 is differentially expressed in colonic tissue between lymphocytic colitis and collagenous colitis, suggesting them of separate disease processes. Finally, patients with alternating IBD and MC episodes represent a diverse group. Among them, the majority demonstrates MC-like miR-31 expression pattern in MC phases, which seems unlikely to support the speculation of MC as an inactive form of IBD. Although the mechanisms deserve further investigation, microRNA is a potentially useful biomarker to differentiate IBD and MC.


Gauge invariant theory for super high resolution Raman images.
The use of a highly localized plasmonic field has enabled us to achieve sub-nanometer resolution of Raman images for single molecules. The inhomogeneous spatial distribution of plasmonic field has become an important factor that controls the interaction between the light and the molecule. We present here a gauge invariant interaction Hamiltonian (GIIH) to take into account the non-uniformity of the electromagnetic field distribution in the non-relativistic regime. The theory has been implemented for both resonant and nonresonant Raman processes within the sum-over-state framework. It removes the gauge origin dependence in the phenomenologically modified interaction Hamiltonian (PMIH) employed in previous studies. Our calculations show that, in most resonant cases, the Raman images from GIIH are similar to those from PMIH when the origin is set to the nuclear charge center of the molecule. In the case of nonresonant Raman images, distinct differences can be found from two different approaches, while GIIH calculations provide more details and phase information of the images. Furthermore, the results from GIIH calculations are more stable with respect to the computational parameters. Our results not only help to correctly simulate the resonant and nonresonant Raman images of single molecules but also lay the foundation for developing gauge invariant theory for other linear and nonlinear optical processes under the excitation of non-uniform electromagnetic field.


HL-A antigens in mummified pre-Columbian tissues.
Tissue extracts of pre-Columbian mummies, from 500 to 2000 years old, were found to inhibit specific antibodies to HL-A. Two-thirds of the specimens tested gave positive results. Patterns of reactions obtained with different antiserums detecting the same antigen were concordant and consistent with known relations between HL-A antigens. The distribution of antigens found was similar to that observed in present-day descendants of the ancient populations studied. Although artifacts due to contaminating substances could have occurred, the reactions resembled in many respects those of HL-A antigens rather than those of nonspecific cross-reacting inhibitors. Development of a technique for HL-A typing of mummified remains may open new possibilities for anthropologic studies.


Incipient sympatric speciation in Midas cichlid fish from the youngest and one of the smallest crater lakes in Nicaragua due to differential use of the benthic and limnetic habitats?
Understanding how speciation can occur without geographic isolation remains a central objective in evolutionary biology. Generally, some form of disruptive selection and assortative mating are necessary for sympatric speciation to occur. Disruptive selection can arise from intraspecific competition for resources. If this competition leads to the differential use of habitats and variation in relevant traits is genetically determined, then assortative mating can be an automatic consequence (i.e., habitat isolation). In this study, we caught Midas cichlid fish from the limnetic (middle of the lake) and benthic (shore) habitats of Crater Lake Asososca Managua to test whether some of the necessary conditions for sympatric speciation due to intraspecific competition and habitat isolation are given. Lake As. Managua is very small (<900 m in diameter), extremely young (maximally 1245 years of age), and completely isolated. It is inhabited by, probably, only a single endemic species of Midas cichlids, Amphilophus tolteca. We found that fish from the limnetic habitat were more elongated than fish collected from the benthic habitat, as would be predicted from ecomorphological considerations. Stable isotope analyses confirmed that the former also exhibit a more limnetic lifestyle than the latter. Furthermore, split-brood design experiments in the laboratory suggest that phenotypic plasticity is unlikely to explain much of the observed differences in body elongation that we observed in the field. Yet, neutral markers (microsatellites) did not reveal any genetic clustering in the population. Interestingly, demographic inferences based on RAD-seq data suggest that the apparent lack of genetic differentiation at neutral markers could simply be due to a lack of time, as intraspecific competition may only have begun a few hundred generations ago.


Investigation of partially coherent vector vortex beams with non-isotropic states of spatial correlation.
In this work, the far-field properties of non-isotropic partially coherent vector vortex beams (PCVVBs) are investigated both theoretically and experimentally. The term non-isotropic signifies that the spatial correlations between the parallel and orthogonal electric field components are distinguishable. It is found that self-orientation and shaping of intensity profile, correlation-induced polarization and depolarization are highly dependent on both the non-isotropic correlation parameters and Poincaré-Hopf index (PHI) of the beam. The simultaneous depolarization and polarization effects are due to the difference in the input correlation parameters that alter the state of polarization (SOP) and degree of polarization (DOP) distributions. The experimental results are in good agreement with the theoretical predictions. The distinguishability of correlation parameters at the source plane leads to significant changes on its intensity profile, DOP, and SOP distributions on far-field propagation, which may found potential applications in beam shaping, detecting and imaging atmospheric lidar, optical imaging and directional transportation where the self-rotation characteristic of beam plays an important role.


NUDT16 is a (deoxy)inosine diphosphatase, and its deficiency induces accumulation of single-strand breaks in nuclear DNA and growth arrest.
Nucleotides function in a variety of biological reactions; however, they can undergo various chemical modifications. Such modified nucleotides may be toxic to cells if not eliminated from the nucleotide pools. We performed a screen for modified-nucleotide binding proteins and identified human nucleoside diphosphate linked moiety X-type motif 16 (NUDT16) protein as an inosine triphosphate (ITP)/xanthosine triphosphate (XTP)/GTP-binding protein. Recombinant NUDT16 hydrolyzes purine nucleoside diphosphates to the corresponding nucleoside monophosphates. Among 29 nucleotides examined, the highest k(cat)/K(m) values were for inosine diphosphate (IDP) and deoxyinosine diphosphate (dIDP). Moreover, NUDT16 moderately hydrolyzes (deoxy)inosine triphosphate ([d]ITP). NUDT16 is mostly localized in the nucleus, and especially in the nucleolus. Knockdown of NUDT16 in HeLa MR cells caused cell cycle arrest in S-phase, reduced cell proliferation, increased accumulation of single-strand breaks in nuclear DNA as well as increased levels of inosine in RNA. We thus concluded that NUDT16 is a (deoxy)inosine diphosphatase that may function mainly in the nucleus to protect cells from deleterious effects of (d)ITP.


Validation of the stratify scale for the prediction of falls among hospitalized adults in a tertiary hospital in Colombia: a retrospective cohort study.
The STRATIFY scale has been implemented as a preventive strategy for predicting the risk of accidental falls among hospitalized adults. However, there is still uncertainty about its accuracy. This study aimed to perform an external validation of the STRATIFY fall prediction scale in hospitalized adults in one tertiary care hospital in Bogotá, Colombia. The study was a retrospective cohort of adult hospitalized patients in a high-level complexity care hospital. The sample selected included admitted patients (age ≥ 18), consecutively by the institution between 2018 and 2020, with an evaluation of the fall risk measured by the STRATIFY score given to each at the time of hospital admission. For assessing the scale's feasibility, its discriminative capability was obtained by calculating sensitivity, specificity, likelihood ratios, predictive values, and area under the ROC curve. The evaluation included 93,347 patient hospital records (mean 56.9 years, 50.2% women). The overall sensitivity score was 0.672 [IC 95% 0.612-0.723], the specificity score was 0.612 [IC 95% 0.605-0.615], and the positive likelihood ratio was 1.73 [IC 95% 1.589-1.891]. The area under the ROC curve was 0.69 [IC 95% 0.66-0.72]. Subgroups of age obtained similar results. Applying the STRATIFY scale at hospital admission resulted in a lower performance of the tool-predict falls in hospitalized patients. It is necessary to implement an individual evaluation of the risk factors for falls in order to structure appropriate care plans to prevent and improve hospital safety.


[Future prospects in developing of dengue vaccines].
Dengue is a viral, febrile and acute disease that causes more morbility than any other infection for Arbovirus. It becomes necessary to implement a safe and effective vaccine that will contribute to the eradication of this disease, especially in endemic areas with very low socio-economic conditions. The only option that exists at the present to control the transmission of the disease is through vector control. So far, there are no effective antiviral drugs for flavivirus such as dengue. Candidates for vaccines for this disease, who are currently in advanced stages of development, are live attenuated vaccine, live attenuated chimerics, inactivated virus, subunits and DNA, in order to mimic the natural infection. The requirements for an ideal vaccine capable of protecting the population at risk should be: free reactogenicity, one which induces protection against all four serotypes of the virus (tetravalent), that offers effective prolonged protection; it also should be highly immunogenic, with few side effects, easy administration, available at low cost and should be administered in one dose, both in adults and in children. Meanwhile, efforts to try to give immunoprophylaxis for this disease continue, in order to design an ideal vaccine allowing the eradication of the disease and the reduction in the number of cases which affects our people every day.


Anti-HIV effects of CD4-Pseudomonas exotoxin on human lymphocyte and monocyte/macrophage cell lines.
CD4(178)-PE40 is a recombinant protein consisting of the HIV envelope glycoprotein-binding region of human CD4 linked to active domains of Pseudomonas aeruginosa exotoxin A. The hybrid toxin selectively kills HIV-infected human T-cell lines and protects against HIV spread in mixtures of uninfected and infected cells. We now report that CD4(178)-PE40 also selectively kills chronically HIV-1-infected cells of monocyte/macrophage lineage. The results provide further support for therapeutic use of this hybrid toxin in the treatment of HIV-infected individuals.


Building the process-drug-side effect network to discover the relationship between biological processes and side effects.
Side effects are unwanted responses to drug treatment and are important resources for human phenotype information. The recent development of a database on side effects, the side effect resource (SIDER), is a first step in documenting the relationship between drugs and their side effects. It is, however, insufficient to simply find the association of drugs with biological processes; that relationship is crucial because drugs that influence biological processes can have an impact on phenotype. Therefore, knowing which processes respond to drugs that influence the phenotype will enable more effective and systematic study of the effect of drugs on phenotype. To the best of our knowledge, the relationship between biological processes and side effects of drugs has not yet been systematically researched. We propose 3 steps for systematically searching relationships between drugs and biological processes: enrichment scores (ES) calculations, t-score calculation, and threshold-based filtering. Subsequently, the side effect-related biological processes are found by merging the drug-biological process network and the drug-side effect network. Evaluation is conducted in 2 ways: first, by discerning the number of biological processes discovered by our method that co-occur with Gene Ontology (GO) terms in relation to effects extracted from PubMed records using a text-mining technique and second, determining whether there is improvement in performance by limiting response processes by drugs sharing the same side effect to frequent ones alone. The multi-level network (the process-drug-side effect network) was built by merging the drug-biological process network and the drug-side effect network. We generated a network of 74 drugs-168 side effects-2209 biological process relation resources. The preliminary results showed that the process-drug-side effect network was able to find meaningful relationships between biological processes and side effects in an efficient manner. We propose a novel process-drug-side effect network for discovering the relationship between biological processes and side effects. By exploring the relationship between drugs and phenotypes through a multi-level network, the mechanisms underlying the effect of specific drugs on the human body may be understood.


Cost-effectiveness of computer-tailored smoking cessation advice in primary care: a randomized trial (ESCAPE).
Smoking remains a significant public health problem and is a leading cause of preventable morbidity and mortality around the world. By combining the behavioral intervention principles used in specialist services with the high reach rates of public health interventions, personal tailored self-help cessation intervention provides a potential economical method for improving reach and effectiveness. Cost-effectiveness analysis is performed alongside a randomized controlled trial to compare the computer-tailored self-help intervention with a generic self-help intervention in smoking cessation. A Markov model was developed to extrapolate lifetime cost-effectiveness by combining trial data with estimates from the literature. In the short term, smokers in the intervention group gained 0.0006 (95% CI = -0.0024 to 0.0036) quality-adjusted life years (QALYs) more than those in the control group at an increased cost of about £9 per person (95% CI = £5-£12). This yielded an incremental cost-effectiveness ratio (ICER) of £14,432/QALY. Precision of the ICER estimates was assessed by 5,000 bootstrapping replications. The probability that the intervention was cost effective was 54% (58%) at a cost-effectiveness threshold of £20,000(£30,000) per QALY. The Markov model showed that simulating lifetime outcomes improved the cost-effectiveness ratio (£9,700/QALY) in favor of the tailored intervention. The intervention would have a 55%-57% chance of being more cost effective than nontailored intervention at the willingness-to-pay threshold of £20,000-30,000/QALY. The computer-tailored intervention appears slightly more likely to be cost effective than the generic self-help intervention in smoking cessation, in both the short term and the long term, but caution is required given the considerable uncertainty surrounding the estimates.


Insights on Structural Characteristics and Ligand Binding Mechanisms of CDK2.
Cyclin-dependent kinase 2 (CDK2) is a crucial regulator of the eukaryotic cell cycle. However it is well established that monomeric CDK2 lacks regulatory activity, which needs to be aroused by its positive regulators, cyclins E and A, or be phosphorylated on the catalytic segment. Interestingly, these activation steps bring some dynamic changes on the 3D-structure of the kinase, especially the activation segment. Until now, in the monomeric CDK2 structure, three binding sites have been reported, including the adenosine triphosphate (ATP) binding site (Site I) and two non-competitive binding sites (Site II and III). In addition, when the kinase is subjected to the cyclin binding process, the resulting structural changes give rise to a variation of the ATP binding site, thus generating an allosteric binding site (Site IV). All the four sites are demonstrated as being targeted by corresponding inhibitors, as is illustrated by the allosteric binding one which is targeted by inhibitor ANS (fluorophore 8-anilino-1-naphthalene sulfonate). In the present work, the binding mechanisms and their fluctuations during the activation process attract our attention. Therefore, we carry out corresponding studies on the structural characterization of CDK2, which are expected to facilitate the understanding of the molecular mechanisms of kinase proteins. Besides, the binding mechanisms of CDK2 with its relevant inhibitors, as well as the changes of binding mechanisms following conformational variations of CDK2, are summarized and compared. The summary of the conformational characteristics and ligand binding mechanisms of CDK2 in the present work will improve our understanding of the molecular mechanisms regulating the bioactivities of CDK2.


MicroRNAs in Head and Neck Cancer.
microRNAs (miRs) are small noncoding single-stranded RNAs, about 19-25 nucleotides long. They have been shown to be capable of altering mRNA expression; thus some are oncogenic or tumour suppressive in nature and are regulated by cellular and epigenetic factors. The molecular pathogenic pathway of many cancers has been modified since the discovery of miRs. Head and neck squamous cell carcinoma (HNSCC), the sixth most common cancer in the world, has recently been associated with infection by the human papillomavirus (HPV). miR expression profiles are altered in the transition from dysplasia to carcinoma, with some changes being specific to the underlying risk factor. This difference is particularly significant in HPV-positive HNSCC where host miRs are modulated by the virus, creating a different profile to HPV-negative HNSCC. Saliva, as an easily collected proximal biofluid containing numerous miRs, presents an attractive noninvasive diagnostic tool in detecting HNSCC and determining prognosis. Furthermore, miRs may play a role in the analysis of surgical margins for residual tumour extension and in the development of novel miR-based therapeutic targets and agents.


Dantrolene for muscle rigidity in progressive supranuclear palsy.
Progressive supranuclear palsy is a degenerative neurological condition with a high level of associated motor symptom burden manifesting in poor postural reflexes, bradykinesia, dystonia and stiffness in the body core and neck. In the light of a paucity in literature exploring pain management in neurodegenerative diseases, the below case report describes the use of dantrolene to successfully relieve distressing widespread dystonia and muscle rigidity refractory to non-pharmacological and pharmacotherapy. To our knowledge, this is the first reported case of dantrolene use for the treatment of refractory muscle rigidity pain in neurodegenerative conditions.


Measures of continuity of care. A register-based correlation study.
In an empirical study using data from a health center in Sweden, correlation coefficients were computed among nine different measures of continuity of care, five of them visit-based and four individual-based. Generally, the correlations were high. This may be due, in part, to the similar behavior of the measures for people making few visits. The correlations were also quite high, however, when the sample was restricted to people with many visits. Several measures display a significant dependence on utilization level. The results suggest that, for general purposes, the measure COC should be preferred among the individual-based measures and fraction-of-care continuity among the visit-based measures. On grounds of flexibility and ease of interpretation, the authors recommend fraction-of-care measures.


Riproximin's activity depends on gene expression and sensitizes PDAC cells to TRAIL.
Riproximin (Rpx) is a type II ribosome inactivating protein, which was investigated for its activity in pancreatic ductal adenocarcinoma (PDAC) in a panel of 17 human and rat PDAC cell lines and in rat pancreatic cancer liver metastasis. Cytotoxicity in response to Rpx was determined by MTT assay, apoptosis by flow cytometry and qRT-PCR for apoptosis related genes, and the modulation of the transcriptome was monitored by micro array analysis. The combination effect of Rpx and TRAIL was assessed by MTT assay. Rpx showed high but varying cytotoxicity in PDAC cells. Based on overall gene expression, the sensitivity of these cells was linked to genes involved in apoptosis. Furthermore, based on the affinity of Rpx for CEA, the expression of carcinoembryonic antigen-related cell adhesion molecule (CEACAM) genes was significantly related to Rpx's cytotoxicity in cells with CEACAM gene expression. Exposure of Suit2-007 cells to Rpx induced the mRNA expression of members of signaling pathways initiating from most death receptors, and down modulation of TRAIL. Apoptosis was increased as shown by FACS analysis. Combination of Rpx with TRAIL resulted in a synergistic cytotoxic effect in human Suit2-007 and rat ASML cells, as evidenced by a 6-fold lower tumor cell survival than expected from an additive combination effect. Treatment of BDX rats bearing intra-portally implanted Suit2-007 cells showed a highly significant anticancer effect and indicated an application of Rpx against pancreatic cancer metastasis to the liver. These data favor further evaluation of Rpx as anticancer agent in PDAC.


Clotting activation after oral anticoagulant therapy discontinuation: a risk factor for recurrent venous thromboembolism.
A hypercoagulable state has been reported in some patients treated for venous thromboembolism (VTE) after oral anticoagulant treatment (OAT) discontinuation. It is unclear whether this is a risk factor for thrombosis recurrence. We investigated 139 patients with VTE and followed them up for a median of 20.5 months (6-90 months) to evaluate whether fragment 1 + 2 (F1 + 2) plasma levels are prognostic for VTE recurrence and to confirm clotting activation after OAT withdrawal. Fourteen patients had recurrences during the follow-up. F1 + 2 was measured the day before OAT withdrawal (T0) and 4 weeks later (T1) and its levels were similar in patients with spontaneous VTE versus those with transient risk factors for VTE. F1 + 2 levels increased from T0 to T1 (P < 0.0005). At T1, F1 + 2 values were significantly higher in patients with recurrence than in those without (P < 0.005). The negative predictive value of normal levels of F1 + 2 at T1 was 95%. In carriers of thrombophilic conditions no correlation was found between F1 + 2 levels and VTE recurrence. The results of this study confirm clotting activation after OAT discontinuation and suggest that higher F1 + 2 plasma levels are an independent risk factor for VTE recurrence.


EPI-001 is a selective peroxisome proliferator-activated receptor-gamma modulator with inhibitory effects on androgen receptor expression and activity in prostate cancer.
The androgen receptor (AR) is a driver of prostate cancer (PCa) cell growth and disease progression. Therapies for advanced PCa exploit AR dependence by blocking the production or action of androgens, but these interventions inevitably fail via multiple mechanisms including mutation or deletion of the AR ligand binding domain (LBD). Thus, the development of new inhibitors which act through non-LBD interfaces is an unmet clinical need. EPI-001 is a bisphenol A-derived compound shown to bind covalently and inhibit the AR NH2-terminal domain (NTD). Here, we demonstrate that EPI-001 has general thiol alkylating activity, resulting in multilevel inhibitory effects on AR in PCa cell lines and tissues. At least one secondary mechanism of action associated with AR inhibition was found to be selective modulation of peroxisome proliferator activated receptor-gamma (PPARγ). These multi-level effects of EPI-001 resulted in inhibition of transcriptional activation units (TAUs) 1 and 5 of the AR NTD, and reduced AR expression. EPI-001 inhibited growth of AR-positive and AR-negative PCa cell lines, with the highest sensitivity observed in LNCaP cells. Overall, this study provides new mechanistic insights to the chemical biology of EPI-001, and raises key issues regarding the use of covalent inhibitors of the intrinsically unstructured AR NTD.


PRIME score for prediction of permanent pacemaker implantation after transcatheter aortic valve replacement.
We sought to produce a simple scoring system that can be applied at clinical visits before transcatheter aortic valve replacement (TAVR) to stratify the risk of permanent pacemaker (PPM) after the procedure. Atrioventricular block is a known complication of TAVR. Current models for predicting the risk of PPM after TAVR are not designed to be applied clinically to assist with preprocedural planning. Patients undergoing TAVR at the University of Colorado were split into a training cohort for the development of a predictive model, and a testing cohort for model validation. Stepwise and binary logistic regressions were performed on the training cohort to produce a predictive model. Beta coefficients from the binary logistic regression were used to create a simple scoring system for predicting the need for PPM implantation. Scores were then applied to the validation cohort to assess predictive accuracy. Patients undergoing TAVR from 2013 to 2019 were analyzed: with 483 included in the training cohort and 123 included in the validation cohort. The need for a pacemaker was associated with five preprocedure variables in the training cohort: PR interval > 200 ms, Right bundle branch block, valve-In-valve procedure, prior Myocardial infarction, and self-Expandable valve. The PRIME score was developed using these clinical features, and was highly accurate for predicting PPM in both the training and model validation cohorts (area under the curve 0.804 and 0.830 in the model training and validation cohorts, respectively). The PRIME score is a simple and accurate preprocedural tool for predicting the need for PPM implantation after TAVR.


Presence of prostate cells in bone marrow biopsies as a sign of micrometastasis in cancer patients.
The presence of prostate cancer cells in bone marrow of patients with clinically localized disease is associated with increased chance of disease recurrence. The presence of prostate specific antigen (PSA) in bone marrow aspirates has been used to indicate the presence of micrometastasis. The aim of this study was to present a prospective study of prostate cancer patients to determine the presence of cells that express PSA in aspirates taken from bone marrow and to compare with bone marrow biopsy samples. Results indicated a significant difference between the frequency of cells detected in bone marrow aspirate and biopsy samples (P<0.0002) when all patients were considered. There was no difference between the frequencies of cells detected in bone marrow aspirate and biopsy of patients analyzed before treatment. However, there was a significant (P<0.003) difference between them after treatment. There was also a significant difference in the frequency of PSA positive cells detected (P<0.005) in Stages I to IV as well as in the frequency of cells detected (P<0.0002) when analyzed according to Gleason score. The present results explain the lack of correlation between positive aspirates and prognosis in numerous clinical cases.


[A family with multiple endocrine neoplasia type I presenting prolactinoma, Zollinger-Ellison syndrome and hyperparathyroidism].
A family of multiple endocrine neoplasia type I with five confirmed cases in three generations is described. All of them have primary hyperparathyroidism in common. The propositus is 51 year-old male. After a year of symptoms of gastroduodenal ulcer, he was found to have elevated levels of serum gastrin and PTH. The serial imaging studies revealed a tumor in pancreatic head, and Zollinger-Ellison syndrome was diagnosed. The gastrin level was reduced into normal range after extirpation of the tumor, but post surgical elevation of Calcium put the patient under parathyroidectomy, which normalized serum PTH and Calcium levels. His two sisters (I and II), the mother of them, and the daughter of sister I, had neither signs nor symptoms until family study showed hypercalcemia in all. Sister I is a 54 year-old female with enlarged parathyroid. The hyperparathyroidism is of chemical type, but no other endocrinological abnormality is found. The Calcium level decreased after parathyroidectomy. Sister II is a 56 year-old female. The only sign was galactorrhea. Serum PTH and Calcium decreased after parathyroidectomy. The prolactinoma was diagnosed by the increased prolactin levels and enhanced mass lesion in sella turcica. Her serum prolactin levels is now within normal range since she is on bromocryptine. The mother of the above three siblings and the daughter of the sister I are now under further study.