Effects of vaginal cylinder position on dose distribution in patients with endometrial carcinoma in treatment of vaginal cuff brachytherapy.
To investigate the impact of different cylinder positions on dosimetry of critical structures in patients with endometrial carcinoma undergoing three-dimensional image-based vaginal cuff brachytherapy (VCB). We delivered VCB at a dose of 4 Gy to a depth of 5 mm in the vaginal cuff of 15 patients using three different cylinder positions (neutral [N], parallel [P], and angled [A]) according to the longitudinal axis of the patient. We analyzed the dose-volume distribution and volumetric variability of the rectum and bladder. We converted the total doses to equivalent doses in 2 Gy (EQD2) using a linear-quadratic model (a/b = 3 Gy). The mean rectum volume for the N, P, and A positions was 68.2 ± 22.7 cc, 79.3 ± 33.7 cc, and 74.2 ± 29.6 cc, respectively. The mean rectum volume for the P position was significantly larger than that for the N position (p = 0.03). Relative to the N position, the A position resulted in a lower total EQD2 in the highest irradiated 2 cc (D2cc; p = 0.001), 1 cc (D1cc; p = 0.004), and 0.1 cc (D0.1cc; p = 0.047) of the rectum. Similarly, the P position resulted in a lower EQD2 in the D2cc (p = 0.018) and D1cc (p = 0.024) of the rectum relative to the N position. In the bladder, the P position resulted in a higher EQD2 in the D2cc relative to the N position (p = 0.02). There was no dosimetric difference between the P and A positions in either the rectum or the bladder. Vaginal cuff brachytherapy in the P and A positions is significantly superior to that in the N position in terms of rectum dosimetry. The bladder dose in the N position is considerably lower than that in the other positions.


Superoxide-Mediated Extracellular Mercury Reduction by Aerobic Marine Bacterium Alteromonas sp. KD01.
Microbial reduction plays a crucial role in Hg redox and the global cycle. Although intracellular Hg(II) reduction mediated by MerA protein is well documented, it is still unclear whether or how bacteria reduce Hg(II) extracellularly without its internalization. Herein, for the first time, we discovered the extracellular reduction of Hg(II) by a widely distributed aerobic marine bacterium Alteromonas sp. KD01 through a superoxide-dependent mechanism. The generation of superoxide by Alteromonas sp. KD01 was determined using 3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide and methyl cypridina luciferin analogue as probes via UV-vis and chemiluminescence detection, respectively. The results demonstrated that Hg(II) reduction was inhibited by superoxide scavengers (superoxide dismutase (SOD) and Cu(NO3)2) or inhibitors of reduced nicotinamide adenine dinucleotide (NADH) oxidoreductases. In contrast, the addition of NADH significantly improved superoxide generation and, in turn, Hg(II) reduction. Direct evidence of superoxide-mediated Hg(II) reduction was provided by the addition of superoxide using KO2 in deionized water and seawater. Moreover, we observed that even superoxide at an environmental concentration of 9.6 ± 0.5 nM from Alteromonas sp. KD01 (5.4 × 106 cells mL-1) was capable of significantly reducing Hg(II). Our findings provide a greater understanding of Hg(II) reduction by superoxide from heterotrophic bacteria and eukaryotic phytoplankton in diverse aerobic environments, including surface water, sediment, and soil.


[Concentration of free and esterified cholecalciferol in the tissues of rats with hypervitaminosis D].
Within 4 hrs after intravenous administration into rats of cholecalciferol maximal concentration of the compound, which was increased depending on the dose used, was found in liver tissue, within 24 hrs--in kidney. Increase in cholecalciferol doses led to an increase in the ratio of the esterified compound to its free form. Content of the esterified cholecalciferol reached the highest level in kidney within 4 hrs and 24 hrs after administration of all the doses of cholecalciferol studied.


Biliary tuberculosis mimicking cholangiocarcinoma: treatment with metallic biliary endoprothesis.
A 58-yr-old patient who presented with obstructive jaundice was evaluated with ultrasonography (US), computed tomography (CT), and percutaneous transhepatic cholangiography (PTC). Diffuse irregular stenosis of the extrahepatic bile ducts and periductal ill-defined soft tissue density along the hepatoduodenal ligament was determined. The patient was originally misdiagnosed with cholangiocarcinoma and, because the extent of disease process made surgical bypass impossible, was treated with a percutaneously inserted metallic stent. Histopathological examination of the endoluminal biopsy revealed ductal tuberculosis (TB). Most of the previous reports in the literature indicated that biliary obstruction was due to enlarged tuberculous lymph nodes compressing the bile duct. To our knowledge, only three cases of biliary stricture due to tuberculous involvement of the bile ducts were reported previously. This case illustrates the importance of tissue diagnosis in all cases of obstructive jaundice to avoid missing rare but curable diseases.


Development of a plant-derived subunit vaccine candidate against hepatitis C virus.
Hepatitis C virus (HCV) is a major cause of acute and chronic hepatitis with over 180 million cases worldwide. Vaccine development for HCV has been difficult. Presently, the virus cannot be grown in tissue culture and there is no vaccine or effective therapy against this virus. In this research, we describe the development of an experimental plant-derived subunit vaccine against HCV. A tobamoviral vector was engineered to encode a consensus sequence of hypervariable region 1 (HVR1), a potential neutralizing epitope of HCV, genetically fused to the C-terminal of the B subunit of cholera toxin (CTB). This epitope was selected from among the amino acid sequences of HVR1 "mimotopes" previously derived by phage display technology. The nucleotide sequence encoding this epitope was designed utilizing optimal plant codons. This mimotope is capable of inducing cross-neutralizing antibodies against different variants of the virus. Plants infected with recombinant tobacco mosaic virus (TMV) engineered to express the HVR1/CTB chimeric protein, contained intact TMV particles and produced the HVR1 consensus peptide fused to the functionally active, pentameric B subunit of cholera toxin. Plant-derived HVR1/CTB reacted with HVR1-specific monoclonal antibodies and immune sera from individuals infected with virus from four of the major genotypes of HCV. Intranasal immunization of mice with a crude plant extract containing the recombinant HVR1/CTB protein elicited both anti-CTB serum antibody and anti-HVR1 serum antibody which specifically bound to HCV virus-like particles. Using plant-virus transient expression to produce this unique chimeric antigen will facilitate the development and production of an experimental HCV vaccine. A plant-derived recombinant HCV vaccine can potentially reduce expenses normally associated with production and delivery of conventional vaccines.


X-ray Investigation of CsPbI3:EuCl3 Infiltrated into Gig-Lox TiO2 Spongy Layers for Perovskite Solar Cells Applications.
In this study, we explore the potential of a blended material comprising CsPbI3:EuCl3 perovskite and Gig-Lox TiO2, a unique transparent spongy material known for its multi-branched porous structure, for application in solar cells. The inclusion of EuCl3 in CsPbI3 serves to stabilize the photoactive γ-phase with a bandgap of 1.75 eV, making it suitable for solar energy conversion in tandem solar cells. Our study applies X-ray-based techniques to investigate the structural properties and interfacial behavior within this blended material, in comparison with a reference perovskite layer deposited on glass. In addition, Spectroscopic ellipsometry is complemented with density functional theory calculations and photoluminescence measurements to elucidate the absorption and radiative emission properties of the blend. Notably, our findings reveal a significant quenching of photoluminescence within the blended material, underscoring the pivotal role of the distributed interfaces in facilitating efficient carrier injection from the CsPbI3:EuCl3 perovskite into the Gig-Lox TiO2 sponge. These findings pave the way for the application of the blend as an Electron Transport Layer (ETL) in semi-transparent perovskite solar cells for tandem and building integrated photovoltaics.


Human cytomegalovirus disrupts constitutive MHC class II expression.
CD8(+) and CD4(+) T lymphocytes are important in controlling human CMV (HCMV) infection, but the virus has evolved protean mechanisms to inhibit MHC-based Ag presentation and escape T lymphocyte immunosurveillance. Herein, the interaction of HCMV with the MHC class II Ag presentation pathway was investigated in cells stably transfected with class II transactivator. Flow cytometry experiments demonstrate that HCMV infection decreases cell-surface MHC class II expression. HCMV down-regulates MHC class II surface expression without a significant effect on class II RNA or steady-state protein levels. SDS-stability and confocal microscopy experiments demonstrate normal levels of steady-state peptide-loaded class II molecules in infected cells and that class II molecules reach late endosomal and HLA-DM positive peptide-loading compartments. However, MHC class II positive vesicles are retained in an abnormal perinuclear distribution. Finally, experiments with a mutant HCMV strain demonstrate that this novel mechanism of decreased MHC class II expression is not mediated by one of the known HCMV immunomodulatory genes. These defects in MHC class II expression combined with previously identified CMV strategies for decreasing MHC class I expression enables infected cells to evade T lymphocyte immunosurveillance.


Dipyridamole myocardial SPECT with low heart rate response indicates cardiac autonomic dysfunction in patients with diabetes.
Because dipyridamole is used to assess heart rate (HR) variability, we investigated whether a low HR response during dipyridamole single photon emission computed tomography (SPECT) in patients with diabetes indicates the presence of cardiac autonomic neuropathy (CAN). Subjects were 61 non-insulin-dependent diabetes patients without perfusion defects, myocardial infarction, or arrhythmia who underwent thallium 201 SPECT imaging. The control group comprised 28 subjects without diabetes. HR was measured during infusion of dipyridamole at a rate of 0.14 mg/kg/min, and peak-baseline ratios of 1.20 or less were defined as low. CAN severity was classified by standard autonomic function tests as severe (n = 22), mild (n = 19), or none (n = 20). HR ratios were significantly attenuated in patients with diabetes compared with those in control subjects (1.22 +/- 0.12 vs 1.32 +/- 0.12, P <.001). Among the patients with diabetes, HR ratios decreased as CAN severity increased from none (1.32 +/- 0.10) to mild (1.23 +/- 0.12, P <.05) to severe (1.13 +/- 0.08, P <.005). There was good correlation between HR ratio and R-R interval ratio to deep breathing and to Valsalva, and patients with low HR ratios showed an attenuated response to both tests (all P <.001). The sensitivity and specificity of HR ratios in the detection of CAN were 77% and 74% for severe CAN and 63% and 90% for mild-to-severe CAN, respectively. In patients with diabetes who have normal dipyridamole SPECT results, an attenuated HR response observed during stress indicates a high likelihood of CAN. Further work that assesses these results in diabetes patients with coronary artery disease is warranted.


Evaluation of urinary bladder function after acute spinal cord injury: an experimental study.
The aim of this study was to evaluate the efficacy of early surgical decompression of acute spinal cord injury through the evaluation of urinary bladder function in rabbits. The study was done with 21 New Zealand male rabbits which were 9 to 12 months in age, and weighed an average of 2438 grams (range 2150 to 3550 g). The animals were assigned into four groups as follows: a control group (n=5), a laminectomy group (n=6), a 15-second compression group (n=5) and a 60-second compression group (n=5). A 60 gram compression force was applied on both compression groups with aneurysm clips. All rabbits were sacrificed seven days postoperatively. Urinary bladder tissues were dissected and in vitro relaxation and contraction tests were performed in organ baths. At the beginning of each experiment, 80 mM KCl was added to the isolated organ bath with no significant difference among all four groups (p>0.05). Carbachol was then added to the organ bath and contraction responses were obtained. Carbachol contraction responses were calculated as the percentage of the 80 mM KCl contraction responses, with compression groups showing significant difference from control and sham-operated groups (p<0.05). Electrical field stimulation responses were obtained for all group preparations at 4, 8, 16, 32 Hz frequencies, and showed significant difference in the 15 and 60-second compression groups (p<0.05). The contractility was assessed using E-max and pD2 values. All groups exhibited same pD2 values. The study demonstrated a slightly better outcome for bladder contractility with early decompression. However, there was no significant difference between early and delayed decompression groups.


Terminal deoxynucleotidyl transferase-mediated formation of protein binding polynucleotides.
Terminal deoxynucleotidyl transferase (TdT) enzyme plays an integral part in the V(D)J recombination, allowing for the huge diversity in expression of immunoglobulins and T-cell receptors within lymphocytes, through their unique ability to incorporate single nucleotides into oligonucleotides without the need of a template. The role played by TdT in lymphocytes precursors found in early vertebrates is not known. In this paper, we demonstrated a new screening method that utilises TdT to form libraries of variable sized (vsDNA) libraries of polynucleotides that displayed binding towards protein targets. The extent of binding and size distribution of each vsDNA library towards their respective protein target can be controlled through the alteration of different reaction conditions such as time of reaction, nucleotide ratio and initiator concentration raising the possibility for the rational design of aptamers prior to screening. The new approach, allows for the screening of aptamers based on size as well as sequence in a single round, which minimises PCR bias. We converted the protein bound sequences to dsDNA using rapid amplification of variable ends assays (RAVE) and sequenced them using next generation sequencing. The resultant aptamers demonstrated low nanomolar binding and high selectivity towards their respective targets.


Surgical treatment of glomus jugulare tumors without rerouting of the facial nerve: an infralabyrinthine approach.
Glomus jugulare tumors are benign lesions located in the jugular foramen that may or may not extend into the middle ear, petrous apex, and upper neck; these growths sometimes invade intradurally. The surgical management of these tumors is a challenge to neurosurgeons and skull base surgeons. Because of their abundant vascularity, deep location, complex anatomy, and difficult surgical approach, their treatment, has been a controversial issue for many years. Despite advancements in nonsurgical techniques, the only treatment with proven efficacy is radical surgical removal. The authors present a series of patients treated with radical removal, in which the feasibility of removing glomus jugulare tumors with low morbidity and a surgical approach limited to tumor removal are discussed. The extent of surgical exposure is tailored with emphasis placed on the routine anterior transposition of the facial nerve. Between May 1997 and March 2004, 24 patients with glomus jugulare tumors were treated; 17 patients were women and seven were men. Their mean age at the time of diagnosis was 50 years (range 18-71 years). The most common symptom was hearing loss in 77%, followed by dysphagia and dysphonia in 55% of patients. In seven patients the clinical presentation was a facial palsy. Radical tumor removal was achieved in 23 patients. An anterior facial nerve transposition was not needed in any case. No surgery-related death was recorded in this series, although one patient died of a pulmonary embolism 70 days after the procedure. A one-stage procedure was performed in 23 patients and a two-stage procedure was used in the other patient. Cerebrospinal fluid leakage occurred in two patients. The lower cranial nerve function was worse in eight patients; however, only one had a new deficit. The facial nerve was preserved in all patients except one, in whom a large intradural tumor caused a temporary facial palsy. In the patients with preoperative facial palsy, the tumor only compressed the nerve in three and it invaded the nerve in four. The nerve was decompressed in the cases with no invasion and a graft was placed in the others. The greater auricular nerve was used as a graft in three and the sural nerve was used in one. On follow-up review, the facial nerve function was House-Brackmann Grade 3 in three patients and Grade 2 in three. After 6 months of follow up with no improvement, one patient was referred for a facial muscle transfer. The surgical technique must be tailored to each case. The authors believe that the standard surgical approach to jugular foramen tumors with anterior transposition of the facial nerve should be avoided, and that the extent of surgical exposure must be tailored to each case based on the extent of the tumor and the clinical symptoms. Lower morbidity rates and radical removal can be achieved with a good surgical plan.


Drug-related predictors of readmission for schizophrenia among patients admitted to treatment for drug use disorders.
Patients with schizophrenia and comorbid drug use disorders (DUD) have a severe course of illness. Despite strong evidence that drug use can exacerbate psychotic symptoms, we have limited knowledge of how specific drugs may increase risk of schizophrenia readmission in this group. This study aimed to assess drug-related predictors of readmission for schizophrenia among a national cohort of patients with a history of schizophrenia admitted to DUD treatment. A record-linkage study was used to assess drug-related factors associated with readmission to mental health treatment for schizophrenia, using a consecutive cohort of 634 patients admitted to DUD treatment between 2000 and 2006 in Danish treatment services and tracked until February 2013 or death, controlling for baseline psychiatric treatment variables. The majority of patients were males (79.8%) and the mean age was 34.7years. Of all patients, 78.7% were readmitted for schizophrenia during follow-up, and 6.8% died without having been readmitted. We found a robust association between use of amphetamine at baseline and elevated risk of readmission, a less robust association between use of cannabis and elevated risk of readmission, and no association with cocaine, opioids, alcohol, benzodiazepines, and MDMA. Furthermore, one or more psychiatric inpatients visit in the year prior to DUD admission was robustly associated with elevated risk of schizophrenia readmission. Use of amphetamine and cannabis are risk markers for schizophrenia readmission among patients with a history of schizophrenia and DUD. Psychiatric history is a predictor of schizophrenia readmission in this patient group.


Effective communication for patient safety: transfer note and Modified Early Warning Score.
To analyze the registry of the Transfer Note (NT) and the emission of the Modified Early Warning Score (MEWS) performed by the nurse in adult patients transferred from the Emergency Service as an effective communication strategy for patient safety. A cross-sectional retrospective study developed at a teaching hospital in the South of Brazil that evaluated 8028 electronic medical records in the year 2017. A descriptive analysis was performed. NT reached the institutional target of 95% in January and February, falling below the target in other months. The MEWS measurement was performed in 85.6% (n = 6,870) of the medical records. Of these patients, 96.8% (n = 6,652) had unchanged MEWS. NT and MEWS are inserted in the work of the nurse, however, actions are needed to qualify patient safety, improving effective communication and therefore reducing the possibility of occurrence of adverse events.


Morphological and kinematic basis of the hummingbird flight stroke: scaling of flight muscle transmission ratio.
Hummingbirds (Trochilidae) are widely known for their insect-like flight strokes characterized by high wing beat frequency, small muscle strains and a highly supinated wing orientation during upstroke that allows for lift production in both halves of the stroke cycle. Here, we show that hummingbirds achieve these functional traits within the limits imposed by a vertebrate endoskeleton and muscle physiology by accentuating a wing inversion mechanism found in other birds and using long-axis rotational movement of the humerus. In hummingbirds, long-axis rotation of the humerus creates additional wing translational movement, supplementing that produced by the humeral elevation and depression movements of a typical avian flight stroke. This adaptation increases the wing-to-muscle-transmission ratio, and is emblematic of a widespread scaling trend among flying animals whereby wing-to-muscle-transmission ratio varies inversely with mass, allowing animals of vastly different sizes to accommodate aerodynamic, biomechanical and physiological constraints on muscle-powered flapping flight.


Conserved epitopes in human and mouse tissue-nonspecific alkaline phosphatase. Second report of the ISOBM TD-9 workshop.
A panel of 19 monoclonal antibodies (MAbs) against human tissue-nonspecific (liver/bone/kidney) alkaline phosphatase (TNAP) was obtained through the ISOBM TD-9 workshop. In the present study, the reactivity of these MAbs has been characterized against mouse TNAP. A mouse embryonic stem cell line, frozen sections of long bones, alkaline phosphatase extracted from mouse bone, and serum were used as the source of TNAP for individual assays. Each MAb was tested for immunoreactivity to mouse TNAP by Western blot analysis, immunohistochemistry and enzyme immunoassay. Antibodies 314 and 315 reacted strongly with mouse TNAP in Western blots, while all other antibodies were negative. By immunohistochemistry, antibodies 314, 315 and 333 produced strong positive staining using frozen sections, while antibody 334 was moderately positive. Enzyme immunoassays indicated that MAb 333 was also able to bind to serum TNAP. These antibodies represent very useful reagents to study the pathophysiological expression of TNAP in mouse tissues and in mouse serum.


Which alcohol control strategies do young people think are effective?
The aims of this study were to examine young people's belief in the effectiveness of various alcohol control strategies and to identify demographic, attitudinal and behavioural correlates of perceived effectiveness. An online questionnaire hosted on a secure server was completed by 1418 men and women aged 16-21 years living in South-East England. It assessed the perceived effectiveness of various alcohol control strategies. Key correlates included sensation seeking, impulsivity, conscientiousness, alcohol outcome expectancies, drink refusal self-efficacy, perceived peer alcohol use and Alcohol Use Disorders Identification Test scores. The most effective strategies were perceived to be enforcing responsible service legislation, strictly monitoring late-night licensed premises and teaching alcohol refusal skills. Greater belief in the effectiveness of alcohol control strategies was expressed by older participants, those who consumed less alcohol and those who expected more negative outcomes from alcohol consumption. The data suggest that in order to increase the perceived effectiveness of alcohol control strategies, we may need to address young people's beliefs about the negative outcomes of alcohol use. Strategies that young people believe are effective may be easier to implement, but this does not imply that unpopular but effective strategies should not be tried.


[Rengalin, a New Efficacious and Safe Antitussive Agent. Results of a Randomized, Comparative, Multicenter Clinical Trial in Patients with Acute Respiratory Tract Infections].
Rengalin is a release-active combination antitussive drug based on antibodies to bradykinin, to histamine and morphine. It acts at various mechanisms of cough reflex by modifying endogenous target molecules and their interaction with receptors. The drug's efficacy, as demonstrated previously in experimental and clinical studies, is mediated by specific release-activity obtained as a result of the production process. Efficacy and safety assessment of rengalin in the treatment of cough induced by acute upper respiratory tract infections (URIs) in comparison with a complex codeine-containing drug (codelac) was performed as part of a multicenter, randomized clinical trial involving 143 patients. All the participants presented with dry/non-productive cough caused by URIs (pharyngitis, laryngitis, tracheitis, tracheobronchitis, bronchitis). The duration of cough varied between 12 hours and 7 days. Rengalin was administered in 73 patients receiving 2 tablets 3 times daily for initial three days, and half reduced doses--for the subsequent four days; codelac was administered in 70 patients who were given 1 tablet 3 times daily for the entire treatment period (7 days). Primary efficacy endpoints were time to cough resolution and reduction in the severity of the cough (scored using a Cough Severity Scale). One patient in Rengalin group and three patients in Codelac group were withdrawn from the study. The article presents treatment outcomes obtained for 139 participants who completed the study in accordance with the protocol (Per Protokol-analysis). The data analysis was based on a non-inferiority (or comparability) statistical design for efficacy endpoints. The antitussive effect of rengalin was significantly comparable (p < 0.025) with that of codelac; the time to complete resolution of cough (both daytime and nocturnal) was 7.2 ± 1.0 days (versus 7.0 ± 1.1 in the group of codelac). Rengalin's efficacy was evidenced by a sufficiently reduced cough severity in the initial few days after treatment onset. As a result of the entire 7-day treatment, the severity score was reduced by 3.1 ± 09 (versus 3.1 ± 1.0 in the group of codelac; p < 0.05), totaling 0.2 ± 0.5 point in both groups at the end of the administration period. The frequent non-productive/dry cough was fully resolved in 76% of patients. All the participants in Rengalin group achieved either convalescent outcomes or significant improvement; none of the patients developed secondary bacterial complications. Positive changes in the patients' state over the week were finally confirmed by evaluating the total quality of life scores, including physical and mental component scores (SF-36 questionnaire), and total sleep quality scores, which were comparative between patients treated with rengalin and codelac (p < 0.025). At the end of the administration period, the effect of rengalin was rated by the physician investigators as 'pronounced'. The Clinical Global Impression Scale-Efficacy Indices (CGI-EI) in the groups of rengalin and codelac were comparable, equating a score of 3.7 ± 0.5 (p < 0.025). The safety outcomes of rengalin treatment were assessed across all 143 randomized patients. The drug's high safety profile was confirmed by the absence of adverse events that could be reliably related to the study treatment, and by monitoring of laboratory variables. Rengalin demonstrated good tolerability and favorable compatibility with other medications for URIs with concomitant pathology. The patients showed 100% treatment compliance. Rengalin is a new efficacious and safe drug indicated for the treatment of URI-induced cough. The severity of daytime and nocturnal cough begins to decrease as soon as on the first day after rengalin administration, with severity reduction observed throughout the whole treatment period. At the completion of the 7-day administration, cough severity is reduced by almost 100% and its changes are comparable with the outcomes of treatment with codelac. By targeting various cough reflex mediators, rengalin enables achieving an antitussive effect in the early days after URI onset (in dry, irritative cough episodes), and a protussive effect at later points of treatment. Rengalin promotes resolution of URI-induced cough without development of secondary bacterial complications.


Arginine-Rich Peptide-Based mRNA Nanocomplexes Efficiently Instigate Cytotoxic T Cell Immunity Dependent on the Amphipathic Organization of the Peptide.
To date, the mRNA delivery field has been heavily dominated by lipid-based systems. Reports on the use of nonlipid carriers for mRNA delivery in contrast are rare in the context of mRNA vaccination. This paper describes the potential of a cell-penetrating peptide containing the amphipathic RALA motif to deliver antigen-encoding mRNA to the immune system. RALA condenses mRNA into nanocomplexes that display acidic pH-dependent membrane disruptive properties. RALA mRNA nanocomplexes enable mRNA escape from endosomes and thereby allow expression of mRNA inside the dendritic cell cytosol. Strikingly, RALA mRNA nanocomplexes containing pseudouridine and 5-methylcytidine modified mRNA elicit potent cytolytic T cell responses against the antigenic mRNA cargo and show superior efficacy in doing so when compared to RALA mRNA nanocomplexes containing unmodified mRNA. RALA's unique sequence and structural organization are vital to act as mRNA vaccine vehicle, as arginine-rich peptide variants that lack the RALA motif show reduced mRNA complexation, impaired cellular uptake and lose the ability to transfect dendritic cells in vitro and to evoke T cell immunity in vivo.


Procurement coordinator support group.
This article describes a support group for procurement coordinators. The group was initiated to help coordinators assist grieving donor families and to cope with their own reactions to stressful events. The theory underlying support groups is presented as a guide to understanding the purpose and process of the group. Issues of concern to coordinators are discussed along with strategies for facilitating group discussions and reducing stress.


The Maintenance of Telomere Length in CD28+ T Cells During T Lymphocyte Stimulation.
Telomerase activity is not readily detected in resting human T lymphocytes, however upon antigen presentation, telomerase is transiently upregulated. Presently, it is not known if telomerase activation is necessary for the proliferation of T cells or for the maintenance of telomere lengths. In this study, we found that telomerase activation is not required for the short- term proliferation of T cells and that telomeres progressively shorten in a heterogeneous population of T cells, even if telomerase is detected. By measuring telomerase activity at the single-cell level using quantitative ddPCR techniques (ddTRAP) and by monitoring changes in the shortest telomeres with more sensitive telomere length measurement assays, we show that only a subset of CD28+ T-cells have robust telomerase activity upon stimulation and are capable of maintaining their telomere lengths during induced proliferation. The study of this T-cell subset may lead to a better understanding on how telomerase is regulated and functions in immune cells.


Comparative efficacy and acceptability of treatments for restless legs syndrome in end-stage renal disease: a systematic review and network meta-analysis.
Restless legs syndrome (RLS) is common in end-stage renal disease (ESRD) patients and impairs health and quality of life significantly. However, the optimal treatment of RLS in ESRD patients is uncertain and less studied compared with idiopathic RLS patients. We conducted a systematic review and network meta-analysis to compare the efficacy and acceptability of treatments for RLS in ESRD patients. Randomized controlled trials (RCTs) by February 2019 in the PubMed, Cochrane Library, Embase and ClinicalTrials.gov were reviewed. RLS severity reduction was treated as treatment efficacy, and adverse events were treated as acceptable. Both outcomes were appraised using a random effects model expressed as standardized mean differences and odds ratios with 95% confidence intervals (CIs), respectively, and were ranked using surface under the cumulative ranking curve (SUCRA) probabilities to obtain a hierarchy of interventions. A total of 12 RCTs were included, comprising 9 interventions and 498 participants. All the interventions significantly improved RLS severity without critical side effects compared with placebo. Gabapentin achieved the greatest decrease of RLS severity [standardized mean difference (SMD) = 1.95, 95% CI 0.81-3.09 (SUCRA: 79.3%)], despite its frequent adverse events [SMD = 0.18, 95% CI 0.02-1.50 (19.9%)]. The combination therapy of exercise plus dopamine agonist had better efficacy [SMD = 1.60, 95% CI 0.08-3.12 (59.8%)] and acceptability [SMD = 1.41, 95% CI 0.01-142.53 (63.9%)] compared with that of vitamin C plus vitamin E [SMD = 1.50, 95% CI 0.47-2.54 (56.6%); SMD = 0.32, 95% CI 0.04-2.86 (32.5%)]. This network meta-analysis supports that gabapentin is the most effective treatment for RLS in ESRD patients. Exercise plus dopamine agonist is a favorable combination therapy concerning side effects. Future large RCTs with long-term treatment outcomes are necessary.


Rate of Listeria monocytogenes shedding from frogs.
Leopard frogs (Rana pipiens) were orally inoculated with Listeria monocytogenes. Most listeriae were shed 3-6 days after inoculation. The evidence indicated that the listeriae did not multiply in frogs and were unable to become established in the gastrointestinal tract.


Broccoli and watercress suppress matrix metalloproteinase-9 activity and invasiveness of human MDA-MB-231 breast cancer cells.
A high dietary intake of cruciferous vegetables has been associated with a reduction in numerous human pathologies particularly cancer. In the current study, we examined the inhibitory effects of broccoli (Brassica oleracea var. italica) and watercress (Rorripa nasturtium aquaticum) extracts on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced cancer cell invasion and matrix metalloproteinase-9 activity using human MDA-MB-231 breast cancer cells. Aberrant overexpression of matrix metalloproteinases, including metalloproteinase-9, is associated with increased invasive potential in cancer cell lines. Our results demonstrate that extracts of broccoli and Rorripa suppressed TPA-induced MMP-9 activity and invasiveness in a concentration dependent manner as determined by zymographic analysis. Furthermore, fractionation of individual extracts followed by liquid chromatography mass spectroscopy analysis (LC-MS) revealed that the inhibitory effects of each vegetable were associated with the presence of 4-methysulfinylbutyl (sulforaphane) and 7-methylsulphinylheptyl isothiocyanates. Taken together, our data indicate that isothiocyanates derived form broccoli and Rorripa inhibit metalloproteinase 9 activities and also suppress the invasive potential of human MDA-MB-231 breast cancer cells in vitro. The inhibitory effects observed in the current study may contribute to the suppression of carcinogenesis by diets high in cruciferous vegetables.


Effects of somatic cell count and stage of lactation on raw milk composition and the yield and quality of Cheddar cheese.
The effects of somatic cell count and stage of lactation on the yield and quality of Cheddar cheese were investigated. Cheese was manufactured in a pilot scale factory using milk of low bulk milk cell count (BMCC) from herds in early (LE) and late (LL) lactation, and milk of high BMCC from herds in early (HE) and late (HL) lactation. The deleterious effect of an elevated BMCC on product yield and quality in late lactation was clear. Cheese made from LL milk was significantly superior to that made from HL milk for most yield and quality characteristics measured. Stage of lactation also affected cheese yield and quality, as evidenced by the lower recovery of fat and poorer flavour score for cheese from LL milk compared with that manufactured from LE milk. The observed differences could be explained largely by differences in raw milk composition. We conclude that the effect of stage of lactation was magnified by an elevated BMCC, and that many of the problems encountered when processing late season milk could be overcome by containing mastitis at this time.


A theoretical foundation for state-transition cohort models in health decision analysis.
Following its introduction over three decades ago, the cohort model has been used extensively to model population trajectories over time in decision-analytic modeling studies. However, the stochastic process underlying cohort models has not been properly described. In this study, we explicate the stochastic process underlying a cohort model, by carefully formulating the dynamics of populations across health states and assigning probability rules on these dynamics. From this formulation, we explicate a mathematical representation of the system, which is given by the master equation. We solve the master equation by using the probability generation function method to obtain the explicit form of the probability of observing a particular realization of the system at an arbitrary time. The resulting generating function is used to derive the analytical expressions for calculating the mean and the variance of the process. Secondly, we represent the cohort model by a difference equation for the number of individuals across all states. From the difference equation, a continuous-time cohort model is recovered and takes the form of an ordinary differential equation. To show the equivalence between the derived stochastic process and the cohort model, we conduct a numerical exercise. We demonstrate that the population trajectories generated from the formulas match those from the cohort model simulation. In summary, the commonly-used cohort model represent the average of a continuous-time stochastic process on a multidimensional integer lattice governed by a master equation. Knowledge of the stochastic process underlying a cohort model provides a theoretical foundation for the modeling method.


Native Mass Spectrometry: Insights and Opportunities for Targeted Protein Degradation.
Native mass spectrometry (nMS) is one of the most powerful biophysical methods for the direct observation of noncovalent protein interactions with both small molecules and other proteins. With the advent of targeted protein degradation (TPD), nMS is now emerging as a compelling approach to characterize the multiple fundamental interactions that underpin the TPD mechanism. Specifically, nMS enables the simultaneous observation of the multiple binary and ternary complexes [i.e., all combinations of E3 ligase, target protein of interest, and small molecule proximity-inducing reagents (such as PROteolysis TArgeting Chimeras (PROTACs) and molecular glues)], formed as part of the TPD equilibrium; this is not possible with any other biophysical method. In this paper we overview the proof-of-concept applications of nMS within the field of TPD and demonstrate how it is providing researchers with critical insight into the systems under study. We also provide an outlook on the scope and future opportunities offered by nMS as a core and agnostic biophysical tool for advancing research developments in TPD.


Identification of odour-active compounds of pasteurised orange juice using multidimensional gas chromatography techniques.
Odour-active compounds present in pasteurised orange juice were identified by gas chromatography-olfactometry (GC-O) employing heart-cut multidimensional GC techniques with olfactometry (O) and mass spectrometry (H/C MDGC-O/MS) and comprehensive two-dimensional gas chromatography-accurate mass time-of-flight MS (GC×GC-accTOFMS). Headspace solid phase microextraction sampling proved to be qualitatively adequate for the analysis of pasteurised orange juice. The GC-O approach distinguished 13 potent odour regions (detection frequency≥3) in the orange juice extract, in which 7 regions were then subjected to detailed identification of the compounds that contribute to the odour, by using higher resolution H/C MDGC-O/MS. This analysis permitted the odour-active peaks to be better resolved on the 2D column, with removal from background matrix, for the seven regions. GC×GC-FID and GC×GC-accTOFMS reveal the overall complexity of the volatile compounds in the product and assisted in assignment of the isolated peaks of the odour-active compounds, confirming the identification in a number of cases. Four aldehydes (hexanal, heptanal, octanal, citral), 2 esters (ethyl butanoate, methyl hexanoate), and 4 monoterpenes (α-pinene, D-limonene, linalool, α-terpineol) were confirmed in accordance with olfactometry assessment in the processed juice. This multi-assessment instrument approach of GC-O, GC×GC, and H/C MDGC provided an effective insight into the processed orange juice aroma.


Modular Homogeneous Chromophore-Catalyst Assemblies.
Photosynthetic reaction center (RC) proteins convert incident solar energy to chemical energy through a network of molecular cofactors which have been evolutionarily tuned to couple efficient light-harvesting, directional electron transfer, and long-lived charge separation with secondary reaction sequences. These molecular cofactors are embedded within a complex protein environment which precisely positions each cofactor in optimal geometries along efficient electron transfer pathways with localized protein environments facilitating sequential and accumulative charge transfer. By contrast, it is difficult to approach a similar level of structural complexity in synthetic architectures for solar energy conversion. However, by using appropriate self-assembly strategies, we anticipate that molecular modules, which are independently synthesized and optimized for either light-harvesting or redox catalysis, can be organized into spatial arrangements that functionally mimic natural photosynthesis. In this Account, we describe a modular approach to new structural designs for artificial photosynthesis which is largely inspired by photosynthetic RC proteins. We focus on recent work from our lab which uses molecular modules for light-harvesting or proton reduction catalysis in different coordination geometries and different platforms, spanning from discrete supramolecular assemblies to molecule-nanoparticle hybrids to protein-based biohybrids. Molecular modules are particularly amenable to high-resolution characterization of the ground and excited state of each module using a variety of physical techniques; such spectroscopic interrogation helps our understanding of primary artificial photosynthetic mechanisms. In particular, we discuss the use of transient optical spectroscopy, EPR, and X-ray scattering techniques to elucidate dynamic structural behavior and light-induced kinetics and the impact on photocatalytic mechanism. Two different coordination geometries of supramolecular photocatalyst based on the [Ru(bpy)3](2+) (bpy = 2,2'-bipyridine) light-harvesting module with cobaloxime-based catalyst module are compared, with progress in stabilizing photoinduced charge separation identified. These same modules embedded in the small electron transfer protein ferredoxin exhibit much longer charge-separation, enabled by stepwise electron transfer through the native [2Fe-2S] cofactor. We anticipate that the use of interchangeable, molecular modules which can interact in different coordination geometries or within entirely different structural platforms will provide important fundamental insights into the effect of environment on parameters such as electron transfer and charge separation, and ultimately drive more efficient designs for artificial photosynthesis.


Sustained expression of coagulation factor IX by modified cord blood-derived mesenchymal stromal cells.
Hemophilia B patients are subject to frequent and spontaneous bleeding caused by a deficiency of clotting factor IX (FIX). Mesenchymal stromal cells (MSCs) have been used in cellular therapies as a result of their immunomodulatory properties, the ability to home to sites of injury and their amenability to various ex vivo modifications, including lentiviral-mediated gene transfer. MSCs were isolated from human umbilical cord blood and differentiated into adipogenic, chondrogenic and osteogenic lineages. A lentiviral DNA vector containing the human FIX gene was generated using traditional restriction enzyme digest and ligation techniques to generate viable replication-incompetent lentiviral particles that were used to transduce MSCs. Quantitative measurement of FIX expression was conducted using an enzyme-linked immunosorbent assay. The over-expression of FIX was sustained in vitro at levels > 4 µg/10(6) cells/24 h and FIX coagulant activity was > 2.5 mIU/10(6) cells/24 h for the 6-week duration of study. Lentiviral modification of cells with a multiplicity of infection of 10 did not adversely affect the potential of cord blood (CB) MSCs to differentiate to adipocytes, chondrocytes and osteoblastic cells, and the expression of functional FIX was sustained after differentiation and was similar to that in nondifferentiated cells. Modification of human CB MSCs with a lentiviral vector resulted in sustained high FIX expression in vitro after differentiation to adipogenic, chondrogenic and osteoblastic cells. These modified MSCs could have applications in cellular therapies for hemophilia B.


Integrating Patient Priorities with Science by Community Engagement in the Kidney Precision Medicine Project.
The Kidney Precision Medicine Project (KPMP) is a multisite study designed to improve understanding of CKD attributed to diabetes or hypertension and AKI by performing protocol-driven kidney biopsies. Study participants and their kidney tissue samples undergo state-of-the-art deep phenotyping using advanced molecular, imaging, and data analytical methods. Few patients participate in research design or concepts for discovery science. A major goal of the KPMP is to include patients as equal partners to inform the research for clinically relevant benefit. The purpose of this report is to describe patient and community engagement and the value they bring to the KPMP. Patients with CKD and AKI and clinicians from the study sites are members of the Community Engagement Committee, with representation on other KPMP committees. They participate in KPMP deliberations to address scientific, clinical, logistic, analytic, ethical, and community engagement issues. The Community Engagement Committee guides KPMP research priorities from perspectives of patients and clinicians. Patients led development of essential study components, including the informed consent process, no-fault harm insurance coverage, the ethics statement, return of results plan, a "Patient Primer" for scientists and the public, and Community Advisory Boards. As members across other KPMP committees, the Community Engagement Committee assures that the science is developed and conducted in a manner relevant to study participants and the clinical community. Patients have guided the KPMP to produce research aligned with their priorities. The Community Engagement Committee partnership has set new benchmarks for patient leadership in precision medicine research.


Structural basis for Scc3-dependent cohesin recruitment to chromatin.
The cohesin ring complex is required for numerous chromosomal transactions including sister chromatid cohesion, DNA damage repair and transcriptional regulation. How cohesin engages its chromatin substrate has remained an unresolved question. We show here, by determining a crystal structure of the budding yeast cohesin HEAT-repeat subunit Scc3 bound to a fragment of the Scc1 kleisin subunit and DNA, that Scc3 and Scc1 form a composite DNA interaction module. The Scc3-Scc1 subcomplex engages double-stranded DNA through a conserved, positively charged surface. We demonstrate that this conserved domain is required for DNA binding by Scc3-Scc1 in vitro, as well as for the enrichment of cohesin on chromosomes and for cell viability. These findings suggest that the Scc3-Scc1 DNA-binding interface plays a central role in the recruitment of cohesin complexes to chromosomes and therefore for cohesin to faithfully execute its functions during cell division.


Family-based association study between NOS-I and -IIA polymorphisms and autism spectrum disorders in Korean trios.
Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder with a strong genetic component and environmental risk factors. Nitric oxide (NO), which is produced by nitric oxide synthase (NOS), may play a role in the development of ASD. We genotyped nine single nucleotide polymorphisms (SNPs) in the NOS-I gene and nine SNPs in the NOS-IIA gene and carried out the transmission disequilibrium test (TDT) and haplotype analysis in 151 Korean ASD trios. We found preferential transmission of the A allele of rs8068149 (P = 0.039) and G allele of rs1060826 (P = 0.035) of NOS-IIA in ASD and the haplotype analysis revealed that the two haplotypes had significant associations (P = 0.014 and 0.031, respectively). The behavioral subdomain score of failure to use nonverbal behaviors to regulate social interaction in Autism Diagnostic Interview-Revised (ADI-R) was significantly higher in subjects with the GG or AG allele in rs1060826 of NOS-IIA compared to those who had the AA allele (P = 0.027). These results provide significant but weak evidence for an association between NOS-IIA and ASD in the Korean population.


Making the Invisible Visible: Identifying and Articulating Culture in Practice-Based Evidence.
This study describes a conceptual tool, labeled the "culture cube," developed to identify and articulate the cultural underpinnings of prevention and early intervention projects in five priority populations (i.e., African American, Asian Pacific Islander, Latino, Native American, and Lesbian, Gay, Bisexual, Transgender, Queer, and Questioning), participating in the California Reducing Disparities Project Phase 2 (CRDP Phase 2). The culture cube was developed for evaluation of these practice-based evidence services (PBEs) for three purposes: (a) to focus attention on revealing and articulating more fully the operative worldview and culturally grounded frameworks underlying PBEs, explicitly identifying the links between cultural beliefs and values, community needs, and intervention design; (b) to guide the methods used to assess and evaluate PBEs so that the outcome indicators and process measures are conceptually consistent, community defined, and culturally centered; and (c) to invite communities to use their own indigenous epistemological frameworks to establish credible evidence. After reviewing the literature in this area and describing the theoretical framework for the culture cube, we describe its development, application, and the response to its use in the initial stages of the California Reducing Disparities Project-Phase 2.


Effect of anesthesia on cardiovascular and renal function in the newborn piglet.
The effects of anesthetic agents, nitrous oxide-oxygen, nitrous oxide-oxygen plus ketamine, pentobarbital sodium and halothane, were determined in piglets 6 to 76 hr of age. After observations of cardiovascular and renal function in the unanesthetized state, the drugs were administered and observations were repeated in the anesthetized state. All of the anesthetics caused a decrease in cardiac output and an increase in total peripheral resistance in the newborn piglet. Both nitrous oxide-oxygen plus ketamine and halothane produced a decrease in heart rate while pentobarbital caused an increase and halothane a decrease in systemic blood pressure. Changes in renal function were minimal. Changes in renal blood flow were inconsistent and renal vascular resistance was not modified by any of the drugs. There was little evidence of redistribution of intrarenal blood flow and no significant changes in glomerular filtration rate or sodium excretion. Plasma renin concentration increased after halothane. It was concluded that changes in renal and cardiovascular function due to anesthesia in the newborn were small. Differences between newborn and adult animals may be related to several factors including the state of development of the autonomic nervous system and species differences used in different studies.


Local smoke-free policy development in Santa Fe, Argentina.
To describe the process of approval and implementation of a comprehensive smoke-free law in the province of Santa Fe, Argentina, between 2005 and 2009. Review of the Santa Fe smoke-free legislation, articles published in local newspapers and documentation on two lawsuits filed against the law, and interviews with key individuals in Santa Fe. Efforts to implement smoke-free policies in Santa Fe began during the 1990s without success, and resumed in 2005 when the provincial Legislature approved the first 100% smoke-free subnational law in Argentina. There was no strong opposition during the discussions within the legislature. As in other parts of the world, pro-tobacco industry interests attempted to block the implementation of the law using well known strategies. These efforts included a controversy media campaign set up, the creation of a hospitality industry association and a virtual smokers' rights group, the introduction of a counterproposal seeking modification of the law, the challenge of the law in the Supreme Court, and the proposal of a weak national bill that would 'conflict' with the subnational law. Tobacco control advocates sought media attention as a strategy to protect the law. Santa Fe is the first subnational jurisdiction in Latin America to have enacted a comprehensive smoke-free policy following the recommendations of the World Health Organization (WHO) Framework Convention on Tobacco Control. After 3 years of implementation, pro-tobacco industry forces failed to undermine the law. Other subnational jurisdictions in Argentina, as well as in Mexico and Brazil are following the Santa Fe example.


Reproduction in upheaval: ethnic-specific fertility responses to societal turbulence in Kazakhstan.
This study contributes to the literature on demographic adjustments to societal crises by examining ethnic-specific probabilities of having first, second, and third marital births in late-twentieth-century Kazakhstan. Discrete-time logit models, employing data from the 1995 and 1999 Kazakhstan Demographic and Health Surveys, are fitted. The results show that the probability of a first birth responded to societal cataclysms of the post-Soviet transition, but this response was most manifest and enduring in the ethnic group that had been most demographically advanced and that also found itself most politically and economically vulnerable. While ethnic differences in the probability of second and third births were generally more pronounced than in the probabilities of first birth, the pace of their post-Soviet decline was relatively uniform across all ethnic groups.


Vacuum ultraviolet photoabsorption spectroscopy of crystalline and amorphous benzene.
We present the first high resolution vacuum ultraviolet photoabsorption study of amorphous benzene with comparisons to annealed crystalline benzene and the gas phase. Vapour deposited benzene layers were grown at 25 K and annealed to 90 K under conditions pertinent to interstellar icy dust grains and icy planetary bodies in our solar system. Three singlet-singlet electronic transitions in solid benzene correspond to the 1B2u, 1B1u and 1E1u states, redshifted by 0.05, 0.25 and 0.51 eV respectively with respect to the gas phase. The symmetry forbidden 1B2u ← 1A1g and 1B1u ← 1A1g transitions exhibit vibronic structure due to vibronic coupling and intensity borrowing from the allowed 1E1u ← 1A1g transition. Additionally the 1B2u ← 1A1g structure shows evidence of coupling between intramolecular vibrational and intermolecular lattice modes in crystalline benzene with Davydov crystal field splitting observed. The optically forbidden 0-0 electronic origin is clearly visible as a doublet at 4.69/4.70 eV in the crystalline solid and as a weak broadened feature at 4.67 eV in amorphous benzene. In the case of the 1B1u ← 1A1g transition the forbidden 0-0 electronic origin is only observed in crystalline benzene as an exciton peak at 5.77 eV. Thicker amorphous benzene samples show diffuse bands around 4.3, 5.0 and 5.4 eV that we tentatively assign to spin forbidden singlet-triplet 3B2u ← 1A1g, 3E1u ← 1A1g and 3B1u ← 1A1g transitions respectively, not previously reported in photoabsorption spectra of amorphous benzene. Furthermore, our results show clear evidence of non-wetting or 'islanding' of amorphous benzene, characterised by thickness-dependent Rayleigh scattering tails at wavelengths greater than 220 nm. These results have significant implications for our understanding of the physical and chemical properties and processes in astrochemical ices and highlight the importance of VUV spectroscopy.


Donor variation in the growth properties and osteogenic potential of human marrow stromal cells.
Human marrow stromal cells (MSCs) were isolated from posterior illiac crest marrow aspirates obtained from 17 healthy donors, ages 19-45 years, with no apparent physical disability. First passage hMSCs exhibited growth rates in vitro that varied up to 12-fold between donors. No correlation between growth rate and the age or gender of the donor was evident (P </= 0.05). When hMSCs were cultured without passage for eight days (subconfluent cultures) or 22 days (confluent cultures) in the absence of any osteogenic agonists, levels of alkaline phosphatase enzyme activity varied 40-fold and 10-fold, respectively, between donors. When exposed to osteo-inductive media, donor populations also showed dramatic differences in levels of bone-specific gene induction. Collectively, these data demonstrate that hMSC cultures are composed of a heterogeneous mixture of cells at various stages of differentiation and with distinct osteogenic potentials. Differences in both growth rate and ALP activity were evident in hMSC cultures established from multiple aspirates obtained over a six month period from the same donors. Therefore, it appears that cellular heterogeneity produced by the method of harvest is propagated within and among different donor populations during culture expansion in vitro.


Effects of age on GABA turnover rates in specific hypothalamic areas in female rats.
During aging in female rats the estrous cycle ceases and the animals develop phases of constant estrous (CE) or constant diestrous (CD) prior to the irreversible transition into anestrous. In young rats, gamma-aminobutyric acid (GABA) is of pivotal importance for the release of GnRH. In the medial-preoptic area (MPO) where the majority of the GnRH perikarya are located in the rat, GABA release decreases at the time of the preovulatory LH surge. The suprachiasmatic nucleus (SCN) contains numerous GABA neurons. Neurochemical signals from this hypothalamic nucleus provide temporal information to GnRH neurons and thereby influence the preovulatory LH surge and the length of estrous cycles. To investigate aging-related changes of the activity of hypothalamic GABAergic neurons, we determined GABA turnover rates in various hypothalamic nuclei of CE and CD rats and compared them to those determined in young estrous (E) or diestrous rats (D1). In old female rats, GABA activity in the MPO was significantly decreased compared to E and D1 rats. A selective increase of GABA turnover rates was observed in the SCN of CE animals. No age-related changes were observed in the other examined brain areas. These data provide the first evidence for alterations in GABAergic activity in specific hypothalamic areas that depend on age and reproductive status. These may cause changes in ability to induce preovulatory LH surges and to maintain regular estrous cyclicity.


Group independent component analysis of MR spectra.
This study investigates the potential of independent component analysis (ICA) to provide a data-driven approach for group level analysis of magnetic resonance (MR) spectra. ICA collectively analyzes data to identify maximally independent components, each of which captures covarying resonances, including those from different metabolic sources. A comparative evaluation of the ICA approach with the more established LCModel method in analyzing two different noise-free, artifact-free, simulated data sets of known compositions is presented. The results from such ideal simulations demonstrate the ability of data-driven ICA to decompose data and accurately extract components resembling modeled basis spectra from both data sets, whereas the LCModel results suffer when the underlying model deviates from assumptions, thus highlighting the sensitivity of model-based approaches to modeling inaccuracies. Analyses with simulated data show that independent component weights are good estimates of concentrations, even of metabolites with low intensity singlet peaks, such as scyllo-inositol. ICA is also applied to single voxel spectra from 193 subjects, without correcting for baseline variations, line-width broadening or noise. The results provide evidence that, despite the presence of confounding artifacts, ICA can be used to analyze in vivo spectra and extract resonances of interest. ICA is a promising technique for decomposing MR spectral data into components resembling metabolite resonances, and therefore has the potential to provide a data-driven alternative to the use of metabolite concentrations derived from curve-fitting individual spectra in making group comparisons.


PP2A inhibition is a common event in colorectal cancer and its restoration using FTY720 shows promising therapeutic potential.
Protein phosphatase 2A (PP2A) is a tumor suppressor that regulates many signaling pathways crucial for cell transformation. In fact, decreased activity of PP2A has been reported as a recurrent alteration in many types of cancer. Here, we show that PP2A is frequently inactivated in patients with colorectal cancer, indicating that PP2A represents a potential therapeutic target for this disease. We identified overexpression of the endogenous PP2A inhibitors SET and CIP2A, and downregulation of regulatory PP2A such as PPP2R2A and PPP2R5E, as contributing mechanisms to PP2A inhibition in colorectal cancer. Moreover, we observed that its restoration using FTY720 impairs proliferation and clonogenic potential of colorectal cancer cells, induces caspase-dependent apoptosis, and affects AKT and extracellular signal-regulated kinase-1/2 activation status. Interestingly, treatment with FTY720 showed an additive effect with 5-fluorouracil, SN-38, and oxaliplatin, drugs used in standard chemotherapy in patients with colorectal cancer. These results suggest that PP2A activity is commonly decreased in colorectal cancer cells, and that the use of PP2A activators, such as FTY720, might represent a potential novel therapeutic strategy in colorectal cancer.


Caffeine and diazepam: separate and combined effects on mood, memory, and psychomotor performance.
The effects of caffeine and diazepam on several mood, cognitive, learning, memory, and psychomotor tasks were investigated in a double-blind study of 108 young healthy adults who were randomly assigned to nine treatments; oral administration of caffeine (0, 3 and 6 mg/kg), diazepam (0, 0.15, and 0.30 mg/kg) and their combinations. Subjects completed a battery of tasks once before and twice after administration of the drugs. Caffeine alone showed no effects on cognitive, learning, and memory performance, but impaired fine motor coordination and increased anxiety and tenseness. Diazepam alone produced sedation, lowered other ratings of subjective moods, and impaired cognitive, learning, and memory performance. The two drugs did not antagonize the effects of each other, except in the symbol cancellation task.


Modeling reaching impairment after stroke using a population vector model of movement control that incorporates neural firing-rate variability.
The directional control of reaching after stroke was simulated by including cell death and firing-rate noise in a population vector model of movement control. In this model, cortical activity was assumed to cause the hand to move in the direction of a population vector, defined by a summation of responses from neurons with cosine directional tuning. Two types of directional error were analyzed: the between-target variability, defined as the standard deviation of the directional error across a wide range of target directions, and the within-target variability, defined as the standard deviation of the directional error for many reaches to a single target. Both between- and within-target variability increased with increasing cell death. The increase in between-target variability arose because cell death caused a nonuniform distribution of preferred directions. The increase in within-target variability arose because the magnitude of the population vector decreased more quickly than its standard deviation for increasing cell death, provided appropriate levels of firing-rate noise were present. Comparisons to reaching data from 29 stroke subjects revealed similar increases in between- and within-target variability as clinical impairment severity increased. Relationships between simulated cell death and impairment severity were derived using the between- and within-target variability results. For both relationships, impairment severity increased similarly with decreasing percentage of surviving cells, consistent with results from previous imaging studies. These results demonstrate that a population vector model of movement control that incorporates cosine tuning, linear summation of unitary responses, firing-rate noise, and random cell death can account for some features of impaired arm movement after stroke.


Updates on cytochrome p450-mediated cardiovascular drug interactions.
Cytochrome P (CYP) 450 is a superfamily of hemoproteins that play an important role in the metabolism of steroid hormones, fatty acids, and many medications. Many agents used for management of cardiovascular diseases are substrates, inhibitors, or inducers of CYP450 enzymes. When two agents that are substrates, inhibitors, or inducers of CYP450 are administered together, drug interactions with significant clinical consequences may occur. This review discusses CYP450-mediated cardiovascular drug interactions as well as noncardiovascular drug interactions that produced significant cardiovascular side effects. The principles in predicting drug interactions are also discussed.


Cimetidine-resistant Zollinger-Ellison syndrome: successful management with ranitidine.
In a patient with the Zollinger-Ellison syndrome who had undergone vagotomy, pyloroplasty and, later, gastrojejunostomy, epigastric pain and stomal ulceration recurred despite the use of high doses (2700 mg/d) of cimetidine. Ranitidine, a new histamine H2-receptor antagonist, reversed all symptoms and healed the stomal ulcer without side effects, thus obviating the need for further surgery. Ranitidine may prove to be the drug of choice in the medical management of patients with the Zollinger-Ellison syndrome.


Elucidating Best Geospatial Estimation Method Applied to Environmental Sciences.
The aim of this study is to assess and identify the most suitable geospatial interpolation algorithm for environmental sciences. The research focuses on evaluating six different interpolation methods using annual average PM10 concentrations as a reference dataset. The dataset includes measurements obtained from a target air quality network (scenery 1) and a sub-dataset derived from a partitive clustering technique (scenery 2). By comparing the performance of each interpolation algorithm using various indicators, the study aims to determine the most reliable method. The findings reveal that the kriging method demonstrates the highest performance within environmental sciences, with a spatial similarity of approximately 70% between the two scenery datasets. The performance indicators for the kriging method, including RMSE (root mean square error), MAE (mean absolute error), and MAPE (mean absolute percentage error), are measured at 3.2 µg/m3, 10.2 µg/m3, and 7.3%, respectively.This study addresses the existing gap in scientific knowledge regarding the comparison of geospatial interpolation techniques. The findings provide valuable insights for environmental managers and decision-makers, enabling them to implement effective control and mitigation strategies based on reliable geospatial information and data. In summary, this research evaluates and identifies the most suitable geospatial interpolation algorithm for environmental sciences, with the kriging method emerging as the most reliable option. The study's findings contribute to the advancement of knowledge in the field and offer practical implications for environmental management and planning.


Comprehensive Metabolomic Profiling and Incident Cardiovascular Disease: A Systematic Review.
Metabolomics is a promising tool of cardiovascular biomarker discovery. We systematically reviewed the literature on comprehensive metabolomic profiling in association with incident cardiovascular disease (CVD). We searched MEDLINE and EMBASE from inception to January 2016. Studies were eligible if they pertained to adult humans; followed an agnostic and/or comprehensive approach; used serum or plasma (not urine or other biospecimens); conducted metabolite profiling at baseline in the context of examining prospective disease; and included myocardial infarction, stroke, and/or CVD death in the CVD outcome definition. We identified 12 original articles (9 cohort and 3 nested case-control studies); participant numbers ranged from 67 to 7256. Mass spectrometry was the predominant analytical method. The number and chemical diversity of metabolites were very heterogeneous, ranging from 31 to >10 000 features. Four studies used untargeted profiling. Different types of metabolites were associated with CVD risk: acylcarnitines, dicarboxylacylcarnitines, and several amino acids and lipid classes. Only tiny improvements in CVD prediction beyond traditional risk factors were observed using these metabolites (C index improvement ranged from 0.006 to 0.05). There are a limited number of longitudinal studies assessing associations between comprehensive metabolomic profiles and CVD risk. Quantitatively synthesizing the literature is challenging because of the widely varying analytical tools and the diversity of methodological and statistical approaches. Although some results are promising, more research is needed, notably standardization of metabolomic techniques and statistical approaches. Replication and combinations of novel and holistic methodological approaches would move the field toward the realization of its promise.


State-dependent FRET reports calcium- and voltage-dependent gating-ring motions in BK channels.
Large-conductance voltage- and calcium-dependent potassium channels (BK, "Big K+") are important controllers of cell excitability. In the BK channel, a large C-terminal intracellular region containing a "gating-ring" structure has been proposed to transduce Ca(2+) binding into channel opening. Using patch-clamp fluorometry, we have investigated the calcium and voltage dependence of conformational changes of the gating-ring region of BK channels, while simultaneously monitoring channel conductance. Fluorescence resonance energy transfer (FRET) between fluorescent protein inserts indicates that Ca(2+) binding produces structural changes of the gating ring that are much larger than those predicted by current X-ray crystal structures of isolated gating rings.


Fecal calprotectin concentration in children affected by SIBO.
Calprotectin is a protein especially expressed in neutrophil cytosol. In the last few years, Fecal calprotectin (FC) turned out to be a direct marker of gastrointestinal inflammation. Because of the simplicity of the method, it has been studied in several gastroenterologic diseases but no data are available about its concentration in children with small intestinal bacterial overgrowth (SIBO), a complex and not well known condition defined by an excessive germs proliferation, especially anaerobic, in the small bowel, and characterized by dyspeptic and malabsorption symptoms. The aim of this study was to evaluate FC values in children with SIBO, comparing to healthy subjects, in order to clarify if an inflammatory process coexists with SIBO. We enrolled fifty-eight children affected by SIBO, as diagnosed by Lactulose Breath Test (LBT). They were assessed for FC values on stool samples. We compared them with a control population of 60 healthy children. In SIBO patients, a median value of 36.0 mg/kg and a mean value +/- SD of 43.0 +/- 31.6 mg/kg were calculated, while in healthy controls the median value was 29.5 mg/kg and the mean value +/- SD was 35.7 +/- 20.7 mg/kg, showing no statistically significant differences between the two groups (p = 0.07). FC values are negative in children affected by SIBO, not differing from those obtained in healthy children, suggesting that no subclinical intestinal inflammation involving neutrophils occurs in patients with higher proliferation of bacteria in the small bowel. The presence of high FC levels in children affected by SIBO might not be caused by bacterial overgrowth itself and, in this case, another cause should be investigated.


The development of the stria vascularis in the human foetus.
The development of the stria vascularis in the human cochlea was studied in step sections of 81 human foetal temporal bones. The stria vascularis primordium can be identified as a ridge of epithelial cells on the lateral wall of the cochlear duct. The first signs of differentiation appear at the 11th week, but it is not until the 17th-18th week that the typical trilaminar structure is observed. The appearance of similar cells with notched nuclei in both marginal and mesenchymal layers at this stage suggests the possibility that some of the intermediate cells may be of epithelial origin. By the 21st week, the overall appearance resembles that of the adult structure. This occurs 1 week after the opening of the tunnel of Corti, and possibly marks the onset of cochlear function.


Adaptation-Induced Blindness Is Orientation-Tuned and Monocular.
We examined the recently discovered phenomenon of Adaptation-Induced Blindness (AIB), in which highly visible gratings with gradual onset profiles become invisible after exposure to a rapidly flickering grating, even at very high contrasts. Using very similar stimuli to those in the original AIB experiment, we replicated the original effect across multiple contrast levels, with observers at chance in detecting the gradual onset stimuli at all contrasts. Then, using full-contrast target stimuli with either abrupt or gradual onsets, we tested both the orientation tuning and interocular transfer of AIB. If, as the original authors suggested, AIB were a high-level (perhaps parietally mediated) effect resulting from the 'gating' of awareness, we would not expect the effects of AIB to be tuned to the adapting orientation, and the effect should transfer interocularly. Instead, we find that AIB (which was present only for the gradual onset target stimuli) is both tightly orientation-tuned and shows absolutely no interocular transfer, consistent with a very early cortical locus.


The taxonomy of a new parvicursorine alvarezsauroid specimen IVPP V20341 (Dinosauria: Theropoda) from the Upper Cretaceous Wulansuhai Formation of Bayan Mandahu, Inner Mongolia, China.
A new parvicursorine alvarezsauroid theropod specimen IVPP V20341 from the Upper Cretaceous Wulansuhai Formation of Bayan Mandahu, Inner Mongolia, China is described. IVPP V20341 appears to be distinguishable amongst alvarezsauroids by possible cervical procoely and relatively larger semi-circular caudal neural canals, but these features are not proposed as autapomorphies because current knowledge of alvarezsauroid necks and tails remains sparse. IVPP V20341 is distinguishable from Linhenykus-the sole parvicursorine at Bayan Mandahu-by the location of the origination points of the anterior caudal transverse processes; in IVPP V20341 this is the anterodorsal corner of the centra, whereas in Linhenykus it is the posterior end of the prezygapophyses. A number of additional tentative differences between IVPP V20341 and Linhenykus are also identified, but cannot be confirmed until further details of anatomical variation along the neck and tail are revealed by future finds. Thus, following the study of IVPP V20341 there are still seven parvicursorine species from the Upper Cretaceous Gobi Basin, but future finds could increase this to eight species.


Relationships of work-related psychosocial risks, stress, individual factors and burnout - Questionnaire survey among emergency physicians and nurses.
Psychosocial risks represent a great challenge for safety and health protection at work in Europe. The purpose of this study has been to determine the relationships of psychosocial risks arising from work, stress, personal characteristics and burnout among physicians and nurses in the Emergency Medical Service (EMS). We performed a cross-sectional study based on a questionnaire survey which contained the Copenhagen Psychosocial Questionnaire (COPSOQ) and Copenhagen Burnout Inventory (CBI). A total of 88 physicians and 80 nurses completed the survey. Physicians demonstrated higher emotional (mean (M) ± standard deviation (SD) = 74.57±16.85) and cognitive (M±SD = 75.95±13.74) demands as compared to nurses. Both groups had high sensory demands and responsibilities at work, in spite of the low degree of their autonomy. The meaning of work, commitment to the workplace, and insecurity at work were high for both groups. Among all participants, stressful behavior and reactions were within the limits of low values (< 40) and coping strategies showed high values (> 60). Personal and patient-related burnout was high for both groups, where physicians were significantly affected by work-related burnout. The influence at work, degree of freedom at work, social support, sense of coherence, mental health, and problem-focused coping are negatively related to work-related burnout. Based on personal factors and coping styles, emergency physicians and nurses are representing a self-selective professional group that meets high work demands, great responsibility, strong commitment and insecurity at work. Burnout of physicians and nurses in the EMS tends to be ignored, although it has severe consequences on their mental and general health. Med Pr 2017;68(2):178-178.


Hereditary C5 deficiency in man. III. Studies of hemostasis and platelet responses to zymosan.
Platelet-rich-plasma from two hemostatically normal individuals, genetically lacking the fifth component of complement (C5), failed to exhibit normal platelet aggregation, or serotonin release, in the presence of zymosan. This abnormality was found to reside in the C5D plasma rather than in the platelets as demonstrated by the inability of the deficient plasma to activate zymosan for the aggregation of washed normal platelets. The defect could be corrected by the addition of normal plasma, normal serlm, or highly purified human C5. A plasma abnormality similar to that found in the C5D individuals was also noted in plasmas deficient in C3, C6, and C7; whereas C8 plasma D behaved normally. These data suggest that this platelet reaction requires late acting C components, perhaps as the C567 complex, bound to the zymosan particles.


Comparison of different protocols for telomere length estimation by combination of quantitative fluorescence in situ hybridization (Q-FISH) and flow cytometry in human cancer cell lines.
The end of eukaryotic chromosomes terminates with nucleoprotein structures called telomeres. They insure several functions including capping the end of the chromosomes, ensuring their stability and protecting them from end-to-end fusion and preventing the activation of the DNA damage checkpoints. A flow-FISH methodology, i.e. quantitative fluorescence in situ hybridization (Q-FISH) in combination with flow cytometry, has been developed in our laboratory in order to estimate telomere length in three human cancer cell lines: K-562 (chronic myelogenous leukaemia), IM-9 (multiple myeloma) and 1301 (T cell lymphoblastic leukaemia). Telomeres were visualised after hybridisation with FITC-labelled PNA (Peptide Nucleic Acid) probes. We evaluated the most critical steps of the flow-FISH protocol to ensure reproducibility. Different methodological set ups were compared. Three fixation procedures (ethanol 80%, methanol 80% and formaldehyde 4%) were tested besides different fixation times (15 min and 60 min) as well as hybridization times (2 h and overnight). For each of these protocols the following parameters were compared: forward scatter (related to the cell size), side scatter (related to the cell granularity), DNA (FL3 and FL4 fluorescence) and PNA content (FL1 fluorescence) using an EPICS XL flow cytometer. Regarding the fixation procedures, methanol proved to be the best, followed by ethanol and formaldehyde, with respect to the efficiency to measure the different parameters cited above. Indeed, fixation using methanol gave the optimal PNA signal compared to using ethanol and formaldehyde in two of the studied cell lines (K-562 and 1301); the difference observed was highly significant in the 1301 cell line. The duration of fixation did not show significant interference in the reproducibility of the results for the three cell lines studied. An overnight hybridization appeared to be more effective when compared to the 2-h hybridization in the case of the K-562 cell line. The most important steps of the flow-FISH technique, namely the fixative procedure, as well as the hybridization and the fixation times, were investigated. Considering the latter, suitable protocols were set up for routine and fast telomere length estimation in the cancer cell lines.


[Proteolytic activity of duodenal mucosa in elderly and senile patients in the presence of duodenal hemorrhage from ulcer].
During investigation of proteolisis of duodenum mucosa in patients of elderly and senile age with duodenal ulcer, complicated by hemorrhage, in terms up 8 for days there was revealed considerable increase of proteolytic activity, especially of high-molecular proteins and collagens, maximum--on the first day of hemorrhage. In all terms the revealed activity of anterior wall of duodenum was higher, than posterior.


[The role of dysembryogenesis in the development of spinal cord tumors].
Experience of treatment of 36 patients with extramedullary spinal cord tumors is summarized. Pigmented macules, papillomas or the spinous processes painfulness while irritation were determined on skin in accordance to tumor localization in the innervation zone of one or two segments.


Simultaneous determination of four phytoecdysteroids by LC-MS/MS: application to a comparative pharmacokinetic study in normal and adjuvant arthritis rats after oral administration of C. officinalis Kuan phytoecdysteroids extract.
C. officinalis Kuan is the dry root of Cyathula officinalis Kuan. Clinically, it is used for fall and flutter injury, rheumatism and arthralgia. Phytoecdysteroids have significant anti-inflammatory effects, and the phytoecdysteroids present in C. officinalis Kuan exhibit potential for treating rheumatoid arthritis.This study first developed a selective, accurate and efficient LC-MS/MS method for 12-day pharmacokinetic studies regarding the simultaneous determination of cyasterone, 25-epi-28-epi-cyasterone, precyasterone and capitasterone from C. officinalis Kuan phytoecdysteroids extract in normal and adjuvant arthritis rats.An Agilent Eclipse Plus C18 RRHD column (1.8 µm, 50mm × 2.1 mm) with a gradient mobile phase consisting of water (A) and acetonitrile (B) was used for analysis. The mass analysis was performed in an Agilent 6430 QQQ-MS mass spectrometer with positive mode multiple reaction monitoring (MRM).The results indicated that the AUC0-t and AUC0-∞ values of the four phytoecdysteroids in adjuvant arthritis rats were different from those in normal rats on the first day, which could provide a helpful reference for pharmacological and toxicological studies, as well as clinical applications of C. officinalis Kuan in the treatment of rheumatoid arthritis.


Hot-Band Anti-Stokes Fluorescence Properties of Alexa Fluor 568.
Hot-band absorption and anti-Stokes emission properties of an organic fluorescent dye, Alexa Fluor 568, were characterized and compared with those of Rhodamine 101. The comparison of the properties (e.g., quantum efficiency, spectral distribution, thermal properties, and fluorescence lifetime) between the two dyes confirms that both dyes undergo the same process when excited in the red spectral region. Possible undesirable crosstalk effects and applications in dSTORM microscopy were demonstrated and discussed.


Physical and textural properties of functional edible protein films from soybean using an innovative 3D printing technology.
Increasing market demand for sustainable, environmentally friendly edible film materials has called for the development of new customizable production methods utilizing emerging technologies such as 3D printing. We hereby report a new method to generate functional edible soy protein isolate films prepared from three types of soybeans (AR-R11-7999, MO-S17-17168, and MO-S17-19874R) using an innovative 3D printing technology. The protein contents in AR-R11-7999, MO-S17-17168, and MO-S17-19874R soybean meals and their corresponding protein isolates were 40.0, 39.1, and 39.9; and 84.5, 84.7, and 87.3 % (w/w, dry basis), respectively. Response surface methodology was used to maximize the tensile and puncture strength and minimize the thickness of the 3D-printed edible films using protein concentration, plasticizer concentration (glycerol), and drying time as the independent variables. The optimized film production conditions were determined as soy protein concentration: 8.91%, plasticizer concentration: 3.00%, and drying time: 3.98 h with a desirability value of 0.7428. The optimized conditions were then successfully verified with the original soybean lot with a nonsignificant difference in physical properties. At the optimized conditions, the 3D-printed edible films using three soybean lots revealed: 0.108-0.114 mm thickness; 14.79-16.07 MPa tensile strength; 6.97-8.20 N puncture strength; 90.81-91.53, -1.89 to -1.31, and 14.85-17.25 were color parameters L*, a*, and b*, respectively; 1.22-1.36 g/cm3 density; and 104.4-105.7% elongation at break ratio (%). PRACTICAL APPLICATION: Edible soy protein films produced by an extrusion-based 3D printing approach are highly customizable and precise, and could be produced at an industrial scale. This newly produced environment-friendly soy protein-based edible film can serve as an alternate packaging to synthetic plastics and reduce the environmental landfill problem while adding value to soybean produced in the mid-south United States.


Long-acting Injectable Risperidone Use in an 11-Years-Old Bipolar Child.
Early-onset bipolar disorder is difficult for child psychiatrists in terms of both diagnosis and treatment. The proper diagnostic evaluation is negatively impacted by the atypical clinical manifestation and rapid cycling pattern of the disease, together with common comorbidity with attention-deficit hyperactivity disorder and anxiety disorder. In addition to poor insight, nonadherence to treatment, poor family coping skills, and insufficient child psychiatric inpatient units make clinicians unsuccessful in following up and treating such patients. Risperidone is a commonly used atypical antipsychotic it has been approved for the treatment of manic and mixed episodes of bipolar disorder even in 10-17-year-old patients, and it is commonly used. It has a long-acting injectable formulation. Studies on its long-acting form in younger children are limited. In this case presentation, the diagnostic procedure in an 11-year old child with bipolar disorder will be presented. Long-acting injectable risperidone use in the case of nonadherence to treatment and observed side effects will be discussed.


PKCζ phosphorylates VASP to mediate chemotaxis in breast cancer cells.
Breast carcinoma (BC) is one of the most common malignant cancers in females, and metastasis remains the leading cause of death in these patients. Chemotaxis plays an important role in cancer cell metastasis and the mechanism of breast cancer chemotaxis has become a central issue in contemporary research. PKCζ, a member of the atypical PKC family, has been reported to be an essential component of the EGF-stimulated chemotactic signaling pathway. However, the molecular mechanism through which PKCζ regulates chemotaxis remains unclear. Here, we used a proteomic approach to identify PKCζ-interacting proteins in breast cancer cells and identified VASP as a potential binding partner. Intriguingly, stimulation with EGF enhanced this interaction and induced the translocalization of PKCζ and VASP to the cell membrane. Further experiments showed that PKCζ catalyzes the phosphorylation of VASP at Ser157, which is critical for the biological function of VASP in regulating chemotaxis and actin polymerization in breast cancer cells. Furthermore, in PKCζ knockdown BC cells, the enrichment of VASP at the leading edge was reduced, and its interaction with profilin1 was attenuated, thereby reducing the chemotaxis and overall motility of breast cancer cells after EGF treatment. In functional assays, PKCζ promoted chemotaxis and motility of BC cells through VASP. Our findings demonstrate that PKCζ, a new kinase of VASP, plays an important role in promoting breast cancer metastasis and provides a theoretical basis for expanding new approaches to tumor biotherapy.


Plio-Pleistocene environmental variability in Africa and its implications for mammalian evolution.
Understanding the climatic drivers of environmental variability (EV) during the Plio-Pleistocene and EV's influence on mammalian macroevolution are two outstanding foci of research in African paleoclimatology and evolutionary biology. The potential effects of EV are especially relevant for testing the variability selection hypothesis, which predicts a positive relationship between EV and speciation and extinction rates in fossil mammals. Addressing these questions is stymied, however, by 1) a lack of multiple comparable EV records of sufficient temporal resolution and duration, and 2) the incompleteness of the mammalian fossil record. Here, we first compile a composite history of Pan-African EV spanning the Plio-Pleistocene, which allows us to explore which climatic variables influenced EV. We find that EV exhibits 1) a long-term trend of increasing variability since ∼3.7 Ma, coincident with rising variability in global ice volume and sea surface temperatures around Africa, and 2) a 400-ky frequency correlated with seasonal insolation variability. We then estimate speciation and extinction rates for fossil mammals from eastern Africa using a method that accounts for sampling variation. We find no statistically significant relationship between EV and estimated speciation or extinction rates across multiple spatial scales. These findings are inconsistent with the variability selection hypothesis as applied to macroevolutionary processes.


[Obesity as a current social problem].
The increasing prevalence of overweight and obesity, the classification of the population whose age is the most favourable for working activities as patients, considerable prevalence of the disease among children and young people, the disease progress and severe complications make obesity one of the most important problems today. The development of obesity and complications is mainly caused by energy imbalance which gets aggravated in subjects manifesting lipid metabolism pathology. Disorders in the character and rhythm of nutrition are most remarkable in cases of obesity complicated by cardiovascular pathology, diseases of the hepatobiliary system, diabetes mellitus, and so forth. Taking into consideration the significance of the problem of obesity the Institute of Nutrition, Academy of Medical Sciences of the USSR, developed a program of measures whose realization will favour the solution of the Food Program.


'DrinkThink' alcohol screening and brief intervention for young people: a qualitative evaluation of training and implementation.
Alcohol Screening and Brief Intervention (ASBI) helps reduce risky drinking in adults, but less is known about its effectiveness with young people. This article explores implementation of DrinkThink, an ASBI co-produced with young people, by health, youth and social care professionals trained in its delivery. A qualitative evaluation was conducted using focus groups with 33 staff trained to deliver DrinkThink, and eight interviews with trained participants and service managers. These were recorded, transcribed and a thematic analysis undertaken. DrinkThink was not delivered fully by health, youth or social care agencies. The reasons for this varied by setting but included: the training staff received, a working culture that was ill-suited to the intervention, staff attitudes towards alcohol which prioritized other health problems presented by young people, over alcohol use. Implementation was limited because staff had not been involved in the design and planning of DrinkThink. Staffs' perceptions of alcohol problems in young people and the diverse cultures in which they work were subsequently not accounted for in the design. Co-producing youth focused ASBIs with the professionals expected to deliver them, and the young people whom they target, may ensure greater success in integrating them into working practice.


Polyadenylate polymerase enzymatic activity in mammary tumor cytosols: A new independent prognostic marker in primary breast cancer.
Polyadenylate polymerase (PAP) is one of the enzymes involved in the formation of the polyadenylate tail of the 3' end of mRNA. High levels of PAP activity were associated with rapidly proliferating cells. Here we evaluate the prognostic value of PAP activity in breast cancer patients. PAP specific activity values were measured by a highly sensitive assay in the tumor cytosols of 228 women with primary breast cancer. The median follow-up period was 58 months. PAP specific activity values ranged from 2.1-39.4 units/mg protein in the breast tumor cytosols, and the activity was correlated with the level of expression of the antigen. An optimal cutoff value of 5.5 units/mg extracted protein was first defined by statistical analysis. PAP status was then compared with other established prognostic factors in terms of relapse-free survival (RFS) and overall survival (OS). PAP activity levels had a tendency to increase with tumor-node-metastasis (TNM) stage and were higher in node-positive patients. Evaluation of the prognostic value of PAP was performed using univariate and multivariate analyses. Univariate analysis showed that PAP-positive patients had a less favorable prognosis for both RFS (relative risk (RR) = 2.35; P < 0.001] and OS (RR = 3.15; P < 0.001). PAP significantly added to the prognostic power for RFS (RR = 2.51; P = 0.0012) and OS (RR = 4.21; P < 0.001) in multivariate analysis, whereas patient age, tumor size, and nodal and ER status remained independent factors for predicting survival. When only node-negative patients were examined, PAP was found to be an independent factor for predicting RFS (RR = 3.68; P = 0.0032) and OS (RR = 4.81; P < 0.001). PAP did not appear to have a prognostic significance for node-positive patients. PAP is a new prognostic factor for early recurrence and death in breast cancer patients. Our results suggest that PAP may be used as an independent unfavorable prognostic factor in node-negative breast cancer patients because there were no significant associations between PAP and the other prognostic indicators evaluated in this group of patients.


UBE2N Promotes Melanoma Growth via MEK/FRA1/SOX10 Signaling.
UBE2N is a K63-specific ubiquitin conjugase linked to various immune disorders and cancer. Here, we demonstrate that UBE2N and its partners UBE2V1 and UBE2V2 are highly expressed in malignant melanoma. Silencing of UBE2N and its partners significantly decreased melanoma cell proliferation and subcutaneous tumor growth. This was accompanied by increased expression of E-cadherin, p16, and MC1R and decreased expression of melanoma malignancy markers including SOX10, Nestin, and ABCB5. Mass spectrometry-based phosphoproteomic analysis revealed that UBE2N loss resulted in distinct alterations to the signaling landscape: MEK/ERK signaling was impaired, FRA1 and SOX10 gene regulators were downregulated, and p53 and p16 tumor suppressors were upregulated. Similar to inhibition of UBE2N and MEK, silencing FRA1 decreased SOX10 expression and cell proliferation. Conversely, exogenous expression of active FRA1 increased pMEK and SOX10 expression, and restored anchorage-independent cell growth of cells with UBE2N loss. Systemic delivery of NSC697923, a small-molecule inhibitor of UBE2N, significantly decreased melanoma xenograft growth. These data indicate that UBE2N is a novel regulator of the MEK/FRA1/SOX10 signaling cascade and is indispensable for malignant melanoma growth. Our findings establish the basis for targeting UBE2N as a potential treatment strategy for melanoma.Significance: These findings identify ubiquitin conjugase UBE2N and its variant partners as novel regulators of MAPK signaling and potential therapeutic targets in melanoma. Cancer Res; 78(22); 6462-72. ©2018 AACR.