Indirect assessment of left ventricular performance in acute myocardial infarction.
One hundred patients admitted to a cardiac care unit had indirect serial determinations of left ventricular systolic ejection times. Patients were divided into groups with transmural infarction, nontransmural infarction, and no infarction, according to clinical, biochemical, and electrocardiographic criteria. Total electromechanical systole and left ventricular ejection time were shortened in acute myocardial infarction, whereas the pre-ejection period and its components, the Q-S1 and isovolumic contraction time intervals, were prolonged. The most abnormal measurements were observed in patients with transmural infarction. Patients with nontransmural infarction demonstrated less severe abnormalities of the systolic ejection times, and the patients without infarction were the least affected. Some of the greatest deviations in the measured intervals were observed in the transmural infarction patients who died. Indirect measurement of left ventricular systolic ejection time is a valuable adjunct in the bedside assessment of left ventricular performance and provides a prognostic index for patients with acute myocardial infarction.


[Decentralization and health system reform: what is their impact on malaria incidence in Colombian municipalities?].
Colombia is one of the Latin-American countries with higher malaria incidence and its control is the responsibility of the departments and municipalities. To assess the effect of decentralization within the context of the Social Security Health System on the incidence of malaria in Colombian municipalities. An ecological trend study was carried out in municipalities which reported at least five cases of malaria in 5 of the 7 years between 1998 and 2004. Information on indicators of decentralization of the municipalities, population with health insurance in either the subsidized or contributive regimes as well as incidence of malaria was requested from the health authorities of the departments and municipalities. Socioeconomic and demographic variables were also collected. The behavior of the malaria rates was assessed in relation to the decentralization status of the municipalities. A repeated measure analysis was performed using the generalized estimating equation. The decentralization status of the municipality (IRR=2.36; 95%CI: 1.57-3.56), its proportion of unmet basic needs (IRR=9.35; 95%CI: 3.66-23.89) and of population younger than 40 years of age (IRR=1.8; 95%CI: 1.13-1.23) were associated with malaria incidence in Colombian municipalities. Decentralization status as well as socioeconomic and demographic factors are associated with increased malaria risk in Colombian municipalities.


Antibody-mediated bacterial adhesion to cytomegalovirus-induced Fc receptors. Potential relationship to secondary infections complicating herpesvirus infections.
Cytomegalovirus (CMV) and other viruses within the herpes group have recently been shown to induce Fc receptors in infected monolayers. We have examined the possibility that such receptors might facilitate the adherence of antibody-coated bacteria to CMV-infected cells. To do this, we infected confluent human embryonic lung (HEL) cell monolayers with CMV (strain AD 169) and then used a double radiolabel assay to measure adherence of Escherichia coli 06 to both infected and control monolayers. We examined infected monolayers 48 h after viral seeding, at which time 30-60% of the cells exhibited characteristic cytopathic changes. We compared the adherence of untreated E. coli 06 with the adherence of E. coli 06 that had been preincubated for 1 h at 37 degrees C with either nonimmune or anti-E. coli 06 antiserum. Pretreatment of the E. coli 06 with specific antiserum significantly enhanced its adherence to CMV-infected, but not to control, monolayers (P less than 0.01 by the Mann-Whitney U test). We did not see such enhancement when we used anti-E. coli 06 antiserum to treat a nontypable E. coli. The augmented adherence of antibody-coated E. coli 06 to CMV-infected monolayers was abrogated by pretreating the monolayers with nonimmune serum or purified Fc fragments, but not by pretreating with IgA, IgM, or 1 mM trypan blue. Preincubating HEL cell monolayers with 100 U/ml human leukocyte interferon for 72 h at 37 degrees C did not affect the adherence of antibody-coated E. coli 06 to the monolayers. To determine if antibody-coated bacteria that adhered to the surface of CMV-infected monolayers might themselves act as receptors for microorganisms with Fc binding potential, we compared the adherence of Cowan strain Staphylococcus aureus to CMV-infected and control monolayers that had been preincubated with antibody-coated E. coli 06. The S. aureus adhered significantly better to the former monolayers (P less than 0.001). These results illustrate a previously unrecognized mechanism by which certain herpesviruses might enhance the adherence of secondary pathogens to nonphagocytic cell populations. Such a mechanism, if active in vivo, might facilitate the colonization of mucosal surfaces by these pathogenic microorganisms, and in this way might contribute to both the reported predisposition of CMV-infected patients to secondary infections and to the high prevalence of S. aureus in the vaginal flora of women with histories of genital herpes.


Comorbidity (psycho) analysed.
Many psychiatric patients seem to suffer from a multitude of psychiatric disorders. The term 'comorbidity' was introduced to characterise that situation. This term, however, is multi-interpretable and thus conceals more than it clarifies. Five interpretations of the concept of comorbidity are discussed, in conjunction with the consequences their validity would have for psychiatric research, in particular for the biological branch. If the term comorbidity continues to be used without further qualification, it will slow down the process of conceptualization of novel research strategies in experimental psychiatry, which is so urgently needed.


Role of extracellular compounds in Cd-sequestration relative to Cd uptake by bacterium Sinorhizobium meliloti.
The role of bacterially derived compounds in Cd(II) complexation and uptake by bacterium Sinorhizobium meliloti wild type (WT) and genetically modified ExoY-mutant, deficient in exopolysaccharide production, was explored combining chemical speciation measurements and assays with living bacteria. Obtained results demonstrated that WT- and ExoY-strains excreted siderophores in comparable amounts, while WT-strain produced much higher amount of exopolysaccharides and less exoproteins. An evaluation of Cd(II) distribution in bacterial suspensions under short term exposure conditions, showed that most of the Cd is bound to bacterial surface envelope, including Cd bound to the cell wall and to the attached extracellular polymeric substances. However, the amount of Cd bound to the dissolved extracellular compounds increases at high Cd(II) concentrations. The implications of these findings to more general understanding of the Cd(II) fate and cycling in the environment is discussed.


Outcomes of patients with right ventricular failure requiring short-term hemodynamic support with the Impella RP device.
Right ventricular failure (RVF) after myocardial infarction, cardiotomy, or left ventricular assist device (LVAD) implantation increases morbidity and mortality. RVF also contributes to prolonged length of hospital stay and higher costs of care. The aim of this study was to evaluate the efficacy and safety of the Impella RP (Abiomed, Danvers, MA) in patients with severe RVF in these clinical settings. This is a prospective cohort study of patients with severe RVF treated with a percutaneous right ventricular assist device (RVAD). Sixty patients with RVF refractory to medical treatment received the Impella RP device at 14 United States institutions as part of the Impella RP pre‒ and post‒market approval studies. The study population included 2 cohorts: Cohort A, patients with RVF post-(LVAD) implantation (n = 31); and Cohort B, patients with RVF post-cardiotomy, heart transplant, or myocardial infarction (n = 29). The primary end-point was survival at 30 days or hospital discharge (whichever was longer). Mean age of patients was 59 ± 15 years; 68% were males, 84% had a history of congestive heart failure, 44% had valvular disease, and 35% had pre-operative renal dysfunction. Patients received an average of 3.4 inotropes/vasopressors before the Impella RP implant. Patients were supported with the Impella RP for 4.0 ± 1.5 (0.5 to 14) days. Hemodynamics improved immediately after initiation of device support, with an increase in cardiac index from 1.9 ± 0.1 to 3.1 ± 0.2 liters/min/m2 (p < 0.001) and a decrease in central venous pressure from 19.0 ± 1 to 13 ± 1 mm Hg (p < 0.001). The overall survival at 30 days (or discharge) was 72%. To the best of our knowledge, this study represents the largest prospective study of patients with life-threatening RVF. Mechanical support with the Impella RP device in patients with RVF resulted in rapid hemodynamic improvement with reversal of shock and favorable survival.


[Use of Escherichia coli recF mutant strains for analyzing the genotoxicity of complex platinum compounds].
The bioluminescence method of assessing SOS response in Escherichia coli cells was applied to test the genotoxicity of five complex platinum compounds: cis-diamminedichloroplatinum, cycloplatam, trans-diamminedichloroplatinum, and two trans-binuclear complexes. Strains AB1157 (pPLS-1) and JC9239 (pPLS-1), a variant of AB1157 carrying the recF143 mutation, were used in the test procedure. SOS response in JC9239 cells was shown to be induced by significantly lower concentrations of cis-DDP and cycloplatam than in AB1157 cells. A drastic increase in cis-DDP toxicity for the JC9239 strain was shown. However, the presence of the recF mutation in JC9239 cells retarded the SOS response induced by trans-platinum compounds. The results obtained indicated that the E. coli recF system plays an essential role in repair and utilization of cis-DDP-DNA adducts.


Identification and characterization of a novel endothelin receptor that binds both ETA- and ETB-selective ligands.
This study demonstrates the presence of a novel endothelin (ET) receptor subtype that displays high affinity for both ETA- and ETB-selective ligands. This subtype has been identified in canine spleen membranes using ETB-selective agonists ET-3, IRL-1620, sarafotoxin 6c (S6c) as well as ETA-selective antagonists BQ123 and related cyclic pentapeptides. Binding of 125I-ET-3 to canine spleen membranes was specific and saturable with an apparent dissociation constant of 130 pM and maximum binding (Bmax) of 240.0 fmol/mg protein. Although the apparent affinities obtained with 125I-ET-1 and 125I-ET-3 were comparable (90 and 130 pM, respectively), the maximum binding obtained with 125I-ET-3 was approximately 35% of that obtained with 125I-ET-1, which indicates that canine spleen possesses both ETA and ETB receptors in the ratio 65:35. Competition binding experiments using 125I-ET-3 and unlabeled ET-1, ET-3, S6c, and IRL-1620 suggested that although ET-1 and ET-3 displayed similar high affinity, S6c and IRL-1620 were 20-300-fold weaker than ET-1 and ET-3 in competing for 125I-ET-3 binding to canine spleen membranes. In addition, BQ123, an ETA-selective antagonist, displaced 125I-ET-3 binding from canine spleen with an IC50 value of 30 nM. Similar profiles were obtained with related cyclic pentapeptides. Electrophysiological studies performed on Xenopus laevis oocytes injected with canine spleen poly(A)+ RNA indicated that the ETB receptor present in these tissues is functional and displays the same pharmacology as that observed in binding studies using these membranes. As a comparison, both binding and functional studies were performed in canine lung and the data indicate that the ETB receptor present in this tissue is similar to that of the cloned human ETB receptor but different from that present in canine spleen. These observations were further confirmed by performing cross-linking experiments on these membranes. Although canine lung and cloned human ETB receptors displayed the same molecular weight bands with similar pharmacology, canine spleen ETB receptors displayed different molecular weight bands and different pharmacology. In addition, the ETB receptors present in canine spleen were also identified in canine bladder, monkey spleen and human spleen. Thus, the data presented in this manuscript provide evidence for the presence of a novel ETB receptor in different tissues as well as different species including human.


Keratoconus staging by decades: a baseline ABCD classification of 1000 patients in the Homburg Keratoconus Center.
This retrospective cross-sectional study aims to analyse the keratoconus (KC) stage distribution at different ages within the Homburg Keratoconus Center (HKC). 1917 corneae (1000 patients) were allocated to decades of age, classified according to Belin's ABCD KC grading system and the stage distribution was analysed. 73 per cent (n=728) of the patients were males, 27% (n=272) were females. The highest KC prevalence occurred between 21 and 30 years (n=585 corneae, 294 patients). Regarding anterior (A) and posterior (B) curvature, the frequency of A was significantly higher than B in all age groups for stage 0, 1 and 2 (A0>B0; A1>B1; A2>B2; p<0.03, Wilcoxon matched-pairs test). There was no significant difference between the number of A3 and B3, but significantly more corneae were classified as B4 than A4 in all age groups (p<0.02). The most frequent A|B combinations were A4|B4 (n=451), A0|B0 (n=311), A2|B4 (n=242), A2|B2 (n=189) and A1|B2 (n=154). Concerning thinnest pachymetry (C), most corneae in all age groups were classified as C0>C1>C2>C3>C4 (p<0.04, Wilcoxon matched-pairs test). For the best distance visual acuity (D), a significantly higher number of corneae were classified as D1 compared to D0 (p<0.008; D1>D0>D2>D3>D4). The stage distributions in all age groups were similar. Early KC rather becomes manifest in the posterior than the anterior corneal curvature whereas advanced stages of posterior corneal curvature coincide with early and advanced stages of anterior corneal curvature. Thus, this study emphasises the necessity of posterior corneal surface assessment in KC as enabled by the ABCD grading system.


Moral distress in acute psychiatric nursing: Multifaceted dilemmas and demands.
In this article, the sources and features of moral distress as experienced by acute psychiatric care nurses are explored. A qualitative design with 16 individual in-depth interviews was chosen. Braun and Clarke's six analytic phases were used. Approval was obtained from the Norwegian Social Science Data Services. Participation was confidential and voluntary. Based on findings, a somewhat wider definition of moral distress is introduced where nurses experiencing being morally constrained, facing moral dilemmas or moral doubt are included. Coercive administration of medicines, coercion that might be avoided and resistance to the use of coercion are all morally stressful situations. Insufficient resources, mentally poorer patients and quicker discharges lead to superficial treatment. Few staff on evening shifts/weekends make nurses worry when follow-up of the most ill patients, often suicidal, in need of seclusion or with heightened risk of violence, must be done by untrained personnel. Provision of good care when exposed to violence is morally challenging. Feelings of inadequacy, being squeezed between ideals and clinical reality, and failing the patients create moral distress. Moral distress causes bad conscience and feelings of guilt, frustration, anger, sadness, inadequacy, mental tiredness, emotional numbness and being fragmented. Others feel emotionally 'flat', cold and empty, and develop high blood pressure and problems sleeping. Even so, some nurses find that moral stress hones their ethical awareness. Moral distress in acute psychiatric care may be caused by multiple reasons and cause a variety of reactions. Multifaceted ethical dilemmas, incompatible demands and proximity to patients' suffering make nurses exposed to moral distress. Moral distress may lead to reduced quality care, which again may lead to bad conscience and cause moral distress. It is particularly problematic if moral distress results in nurses distancing and disconnecting themselves from the patients and their inner selves.


Neoadjuvant cemiplimab and surgery for stage II-IV cutaneous squamous-cell carcinoma: follow-up and survival outcomes of a single-arm, multicentre, phase 2 study.
We previously reported rates of pathological complete responses (51% [95% CI 39-62] per independent central review, the primary endpoint) and major pathological responses (13% per independent central review, a secondary endpoint) to neoadjuvant cemiplimab (an anti-PD-1 inhibitor) among 79 patients with locoregionally advanced, resectable cutaneous squamous cell carcinoma. Here, we present follow-up data, including event-free, disease-free, and overall survival. This single-arm, multicentre, phase 2 study included patients aged 18 years or older with resectable stage II-IV (M0) cutaneous squamous cell carcinoma and Eastern Cooperative Oncology Group performance status of 0 or 1. Patients received up to four planned doses of neoadjuvant cemiplimab 350 mg intravenously every 3 weeks followed by curative-intent surgery. After surgery, per investigator discretion, patients received either adjuvant cemiplimab for up to 48 weeks, radiotherapy, or observation alone. Secondary endpoints included in this follow-up analysis are event-free survival, disease-free survival, and overall survival, all summarised using the Kaplan-Meier method. Activity and safety endpoints were analysed for all enrolled patients who received at least one dose of neoadjuvant cemiplimab. In this report, safety data are reported for all patients who received at least one dose of adjuvant cemiplimab. This trial is registered with ClinicalTrials.gov, NCT04154943, has completed enrolment and follow-up is ongoing. Between March 20, 2020, and July 8, 2021, 79 patients were enrolled. Median age was 73 years (IQR 66-81), 67 (85%) patients were male, 12 (15%) were female, 69 (87%) were White, one was Asian (1%), one was other race (1%), and race was not reported for eight (10%). As of data cutoff (Dec 1, 2022), median follow-up was 18·7 months (IQR 15·6-22·1) for all 79 patients. Among 70 patients who had surgery, 65 (93%) had post-surgical management data: 32 (49%) of 65 were observed postoperatively, 16 (25%) received adjuvant cemiplimab, and 17 (26%) received adjuvant radiotherapy. 11 (14%) of 79 patients had event-free survival events, with an estimated 12-month event-free survival of 89% (95% CI 79-94) for all patients. None of 40 patients who had a pathological complete response and one (10%) of ten patients with major pathological response had recurrence. Six (9%) of 70 patients who completed surgery had a disease-free survival event, with an estimated 12-month disease-free survival of 92% (95% CI 82-97). Nine (11%) of 79 patients died, with an estimated 12-month overall survival for all patients of 92% (95% CI 83-96). Four (25%) of 16 patients who received adjuvant cemiplimab treatment had grade 3 adverse events, including one (6%) who had increased blood potassium, one (6%) who had traumatic limb amputation, and two who had serious adverse events (one [6%] cardiomyopathy and one [6%] hypophysitis). There were no grade 4 adverse events or treatment-related deaths. For patients with resectable stage II-IV cutaneous squamous cell carcinoma, neoadjuvant cemiplimab followed by surgery might be a potential treatment option, addressing a substantial unmet need. Regeneron Pharmaceuticals and Sanofi.


Dissolution rate of zinc-containing beta-tricalcium phosphate ceramics.
The dissolution rates of ceramic tricalcium phosphate (TCP) and zinc-containing tricalcium phosphate (ZnTCP) ceramics at pH 5.5 decrease with increasing zinc content. The relative dissolution rates of ceramic ZnTCP with zinc contents of 0.5 and 1.0 mol% are more than three times higher than the relative resorbed volumes of these ZnTCP ceramics which were reported previously. The dissolution rates are likely controlled by only one mechanism, that is polynucleation, in the zinc content range from 0 to 1.0 mol%. Since the surface area of ceramic TCP or ZnTCP is much lower than that of powdered TCP or ZnTCP, the initial dissolution flux of ceramic TCP or ZnTCP remains unchanged for a longer period than that of powdered TCP. As a result, no change in dissolution kinetics is expected during one cycle of osteoclastic resorption. Since neither the dissolution rate nor the change in dissolution kinetics accounts for the previously reported drastic reduction in osteoclastic resorption associated with ZnTCP, it is concluded that the drastic reduction in resorption arises from the suppressive effects of ZnTCP on osteoclast formation or osteoclastic activity.


Etiology of Uveitis in Upper Egypt.
To report the causes of uveitis in a referral ocular inflammation clinic in Upper Egypt. Retrospective medical chart review of all uveitis cases visiting a referral uveitis clinic during the period between January 2015 and January 2020. A total of 982 patients were included. Uveitis was bilateral in 51.7% of the patients. Anterior uveitis was the most common type, followed by posterior uveitis, affecting 34.4% and 25.6% of the study cohort, respectively. About one-third of our patients were beneath the age of 18, and among that group, no specific etiology of uveitis could be determined in about a quarter of the patients by the end of the study period, and juvenile idiopathic arthritis was the most common disease entity. In conclusion, the present report attempted to illustrate the most common causes of uveitis in Upper Egypt. Tuberculosis followed by sarcoidosis were the two leading causes of uveitis in our group of patients.


Australian Cerebral Palsy Child Study: protocol of a prospective population based study of motor and brain development of preschool aged children with cerebral palsy.
Cerebral palsy (CP) results from a static brain lesion during pregnancy or early life and remains the most common cause of physical disability in children (1 in 500). While the brain lesion is static, the physical manifestations and medical issues may progress resulting in altered motor patterns. To date, there are no prospective longitudinal studies of CP that follow a birth cohort to track early gross and fine motor development and use Magnetic Resonance Imaging (MRI) to determine the anatomical pattern and likely timing of the brain lesion. Existing studies do not consider treatment costs and outcomes. This study aims to determine the pathway(s) to motor outcome from diagnosis at 18 months corrected age (c.a.) to outcome at 5 years in relation to the nature of the brain lesion (using structural MRI). This prospective cohort study aims to recruit a total of 240 children diagnosed with CP born in Victoria (birth years 2004 and 2005) and Queensland (birth years 2006-2009). Children can enter the study at any time between 18 months to 5 years of age and will be assessed at 18, 24, 30, 36, 48 and 60 months c.a. Outcomes include gross motor function (GMFM-66 & GMFM-88), Gross Motor Function Classification System (GMFCS); musculoskeletal development (hip displacement, spasticity, muscle contracture), upper limb function (Manual Ability Classification System), communication difficulties using Communication and Symbolic Behaviour Scales-Developmental Profile (CSBS-DP), participation using the Paediatric Evaluation of Disability Inventory (PEDI), parent reported quality of life and classification of medical and allied health resource use and determination of the aetiology of CP using clinical evaluation combined with MRI. The relationship between the pathways to motor outcome and the nature of the brain lesion will be analysed using multiple methods including non-linear modelling, multilevel mixed-effects models and generalised estimating equations. This protocol describes a large population-based study of early motor development and brain structure in a representative sample of preschool aged children with CP, using direct clinical assessment. The results of this study will be published in peer reviewed journals and presented at relevant international conferences. Australia and New Zealand Clinical Trials Register (ACTRN1261200169820).


Carotid body tumors: a subject review and suggested surgical approach.
Carotid body tumors are a rare but potentially difficult surgical entity. Their pathology, physiology, and natural history are reviewed along with surgical results reported in the literature. A surgical approach for removal of these tumors is presented which differs significantly from the recommended techniques in that emphasis is placed on intraoperative monitoring of cerebral blood flow, the the selective use of shunts, a tumor-adventitial plane of dissection, preservation of the carotid artery complex, and mobilization of the parotid gland. Thirteen cases using these techniques are reviewed. The mortality rate and the incidence of cerebrovascular sequelae were both 0%. The major morbidity consisted of injury to the lower cranial nerves in five patients (39%) with tumors larger than 5 cm in length.


Apelin inhibits adipogenesis and lipolysis through distinct molecular pathways.
Apelin is an adipokine secreted by adipocytes. Co-expression of apelin and apelin receptor (APJ) in adipocytes implies the autocrine regulations of apelin on adipocyte functions through yet unknown molecular mechanisms. In the present study, we provide evidence that apelin, through its interaction with APJ receptor, inhibits adipogenesis of pre-adipocytes and lipolysis in mature adipocytes. The detailed molecular pathways underlying apelin signaling is proposed based on our experimental observations. Specifically, we show that apelin suppresses adipogenesis through MAPK kinase/ERK dependent pathways. And by preventing lipid droplet fragmentation, apelin inhibits basal lipolysis through AMP kinase dependent enhancement of perilipin expression and inhibits hormone-stimulated acute lipolysis through decreasing perilipin phosphorylation. Apelin induced decrease of free fatty acid release can be attributed to its dual inhibition on adipogenesis and lipolysis. This study suggests that the autocrine signaling of apelin may serve as a novel therapeutic target for obesity and other metabolic disorders.


COQ6 mutations in human patients produce nephrotic syndrome with sensorineural deafness.
Steroid-resistant nephrotic syndrome (SRNS) is a frequent cause of end-stage renal failure. Identification of single-gene causes of SRNS has generated some insights into its pathogenesis; however, additional genes and disease mechanisms remain obscure, and SRNS continues to be treatment refractory. Here we have identified 6 different mutations in coenzyme Q10 biosynthesis monooxygenase 6 (COQ6) in 13 individuals from 7 families by homozygosity mapping. Each mutation was linked to early-onset SRNS with sensorineural deafness. The deleterious effects of these human COQ6 mutations were validated by their lack of complementation in coq6-deficient yeast. Furthermore, knockdown of Coq6 in podocyte cell lines and coq6 in zebrafish embryos caused apoptosis that was partially reversed by coenzyme Q10 treatment. In rats, COQ6 was located within cell processes and the Golgi apparatus of renal glomerular podocytes and in stria vascularis cells of the inner ear, consistent with an oto-renal disease phenotype. These data suggest that coenzyme Q10-related forms of SRNS and hearing loss can be molecularly identified and potentially treated.


Multivalent adhesion molecule 7 clusters act as signaling platform for host cellular GTPase activation and facilitate epithelial barrier dysfunction.
Vibrio parahaemolyticus is an emerging bacterial pathogen which colonizes the gastrointestinal tract and can cause severe enteritis and bacteraemia. During infection, V. parahaemolyticus primarily attaches to the small intestine, where it causes extensive tissue damage and compromises epithelial barrier integrity. We have previously described that Multivalent Adhesion Molecule (MAM) 7 contributes to initial attachment of V. parahaemolyticus to epithelial cells. Here we show that the bacterial adhesin, through multivalent interactions between surface-induced adhesin clusters and phosphatidic acid lipids in the host cell membrane, induces activation of the small GTPase RhoA and actin rearrangements in host cells. In infection studies with V. parahaemolyticus we further demonstrate that adhesin-triggered activation of the ROCK/LIMK signaling axis is sufficient to redistribute tight junction proteins, leading to a loss of epithelial barrier function. Taken together, these findings show an unprecedented mechanism by which an adhesin acts as assembly platform for a host cellular signaling pathway, which ultimately facilitates breaching of the epithelial barrier by a bacterial pathogen.


Shaping ability of ProFile and K3 rotary Ni-Ti instruments when used in a variable tip sequence in simulated curved root canals.
To compare the shaping ability of ProFile and K3 rotary Ni-Ti instruments when used in a variable tip sequence in simulated curved root canals with different curvature and radius. ProFile or K3.06 taper instruments were used to prepare simulated canals of 20 degrees curvature and 5 mm radius (n = 10) and 30 degrees curvature and 3 mm radius canals (n = 10) in resin blocks. All canals were prepared to an apical size 40 at 0.5 mm from the canal terminus using a variable tip crown-down sequence. Pre- and postinstrumentation digital images were recorded, and an assessment of the canal shape was determined using a computer image analysis program. The material removal from the inner and outer wall of the canal was measured at 28 measuring points, beginning 0.5 mm from the end-point of the canal and the data compared using the Mann-Whitney U-test. In 20 degrees and 30 degrees canals both instruments significantly removed more (P < 0.05) material on the outer wall than the inner wall in the apical half of the canal. For ProFile files there was no significant difference in the amount of material removed on the outer canal wall between the 20 degrees and 30 degrees canals. However, in the K3 groups significantly more (P < 0.05) outer canal wall was removed in the apical area in 20 degrees canals. When comparing both instruments the results showed that in 20 degrees canals K3 instruments removed more outer and inner canal wall than ProFile instruments (P < 0.05) but that there was no significant difference (P > 0.05) between the instruments in 30 degrees canals. Within the limitation of this study, both rotary nickel-titanium instruments prepared a well-shaped root canal with minimal canal transportation.


A Pilot Study of Blood Pressure Monitoring After Cardiac Surgery Using a Wearable, Non-invasive Sensor.
Background: Continuous blood pressure (BP) measurement in intensive care units is based on arterial line (AL) transducers, sometimes associated with clinical complications. Our objective was to evaluate continuous BP measurements obtained from a non-invasive, wireless photoplethysmography (PPG)-based device using two distinct configurations (wristwatch and chest-patch monitors) compared to an AL. Methods: In this prospective evaluation study, comparison of the PPG-based devices to the AL was conducted in 10 patients immediately following cardiac surgery. Pulse rate (PR), systolic BP (SBP), diastolic BP (DBP), and mean arterial pressure (MAP) were recorded using both the AL and the PPG-based devices simultaneously for an average of 432 ± 290 min starting immediately after cardiac surgery. Bland-Altman plots and Pearson's correlations were used to assess the accuracy and degree of agreement between techniques. Results: A total of ~4,000 data points were included in the final analysis. AL measurements for PR, SBP, DBP and MAP were significantly (p < 0.001) and strongly correlated with both the wristwatch (r = 0.99, r = 0.94, r = 0.93 and r = 0.96, respectively) and the chest-patch (r = 0.99, r = 0.95, r = 0.93 and r = 0.95, respectively) monitors. Both configurations showed a marginal bias of <1 mmHg for BP measurements and <1 beat/min for PR [95% limits of agreement -3,3 beat/min; BP measurements: (-6)-(-10), 6-10 mmHg] compared to AL measurements. Conclusion: The PPG-based devices offer a high level of accuracy for cardiac-related parameters compared to an AL in post-cardiac surgery patients. Such devices could provide advanced monitoring capabilities in a variety of clinical settings, including immediate post-operative and intensive care unit settings. Clinical Trial Registration:www.clinicaltrials.gov, NCT03603860.


[The finances and statistics in materials of the G. E. Rhein Commission].
The article analyses financial and statistical aspects of materials from the Interdepartmental Commission for the Revision of Medical Legislation (the G. E. Rhein Commission). The brief analysis of structure, composition and mechanisms of functioning of the Commission is presented. The specificity of the Commission materials is discussed. The information sources are established which were used as background for statistical calculations of the Commission. The calculations themselves are briefly discussed. The particular attention is paid to methods of calculating financial losses in the Russian Empire due to diseases. The study focuses on state health statistics projects and on the financial foundations of the new health care system as well as on their perception by the participants of the Pirogov Society. The data obtained by the Commission was used for devising plans for optimization and centralization of health care, according to which sanitary statistics were to be subordinated to the state. However, financial support to local self-government bodies for sanitary needs was mentioned. Although the physicians from the Pirogov Society had critical attitude to most ideas of the Commission, in the matter of financing they were not only agreed upon with main proposals of the Commission but were also ready to go even further.


Vascular angiotensin-converting enzyme expression regulates local angiotensin II.
We tested the hypothesis that changes in angiotensin-converting enzyme (ACE) gene expression can regulate the rate of local vascular angiotensin II (Ang II) production. We perfused isolated rat hindlimbs with an artificial medium and infused renin and Ang I via the perfusate. Ang I and II were measured by radioimmunoassay. We then increased ACE gene expression and ACE levels in the rat aorta by producing two-kidney, one clip (2K1C) hypertension for 4 weeks. Gene expression was measured by RNAse protection assay, and ACE activity in the vessel wall was measured by the Cushman-Cheung assay. Angiotensin I infusion at 1, 10, 100, and 1000 pmol/mL led to 371 +/- 14 (+/-SEM), 3611 +/- 202, 44,828 +/- 1425, and 431,503 +/- 16,439 fmol/mL Ang II released, respectively, from the hindlimbs (r = .98, P < .001). Thus, the conversion rate did not change across four orders of magnitude, and the system was not saturable under these conditions. In 2K1C hindlimbs, Ang I infusion (0.5 pmol/mL) resulted in increased Ang II generation (157 +/- 16 versus 123 +/- 23 fmol/mL, P = .014 at minute 10) compared with controls. ACE gene expression and ACE activity were increased in 2K1C hindlimbs compared with controls (36 +/- 4 versus 17 +/- 1 mU/mg protein, P < .001). Ang II degradation in the two groups did not differ. To investigate the conversion of locally generated Ang I, we infused porcine renin (0.5 milliunits per mL) into 2K1C and control hindlimbs. Despite markedly higher Ang I release in sham-operated than in 2K1C rats (71 +/- 8 versus 37 +/- 6 pmol/mL, P = .008 at minute 12), Ang II was only moderately increased (36 +/- 3 versus 25 +/- 6 pmol/mL, P = .12 at minute 12). This difference between 2K1C rats and controls reflected a higher rate of conversion in 2K1C rats. Thus, Ang I conversion in the rat hindlimb is linear over a wide range of substrate concentrations and occurs at a fixed relationship. Nevertheless, increased ACE gene expression and ACE activity in the vessel wall lead to an increase in the conversion of Ang I to Ang II. We conclude that local ACE gene expression and ACE activity can influence the local rate of Ang II production.


Homozygous familial hypercholesterolemia in Italy: Clinical and molecular features.
Homozygous familial hypercholesterolemia (HoFH) is a rare genetic disorder characterized by extremely elevated plasma levels of low density lipoprotein cholesterol (LDL-C) and high risk of premature atherosclerotic cardiovascular disease (ASCVD). HoFH is caused by pathogenic variants in several genes, such as LDLR, APOB and PCSK9, responsible for autosomal dominant hypercholesterolemia (ADH), and LDLRAP1 responsible for autosomal recessive hypercholesterolemia (ARH). Aim of this study was the review of the clinical and molecular features of patients with HoFH identified in Italy from 1989 to 2019. Data were collected from lipid clinics and laboratories, which had performed molecular diagnosis in suspected HoFH. Clinical data included baseline lipid levels and ASCVD events. A total of 125 subjects with ADH were identified, of whom 60 were true homozygotes, 58 compound heterozygotes and 7 double heterozygotes for LDLR (likely) pathogenic variants. Compared with compound heterozygotes, true homozygotes showed a more severe lipid phenotype and more ASCVD events. ADH carriers of LDLR negative variants (R-NEG) presented with a more aggressive phenotype, as compared to carriers of LDLR defective variants (R-DEF). Kaplan-Meier analysis showed that the median age of ASCVD event-free survival was 25 years of age in R-NEG as opposed to 50 years of age in R-DEF patients. A total of 66 patients with ARH were also identified, of whom 46 were homozygotes and 20 compound heterozygotes. The phenotypic features of ARH patients were similar to those of R-DEF/ADH patients. Overall, 45% ADH patients and 33% ARH patients did not meet the classic diagnostic criteria for HoFH. In our cohort, the phenotypic variability of HoFH was dependent on the candidate gene involved and the functional impact of its variants on the LDL receptor pathway.


Incomplete presentations in a series of 37 children with Kawasaki disease: the role of the pediatric ophthalmologist.
Kawasaki disease (KD) is an acute and sometimes fatal febrile vasculitis of childhood, the presenting signs of which include conjunctival vessel dilatation and iridocyclitis. Consultation with a pediatric ophthalmologist is helpful for early recognition of the disease, especially in identifying "incomplete" cases, ie, those which lack all of the classical, systemic signs. After encountering a case of incomplete KD in which diagnosis was delayed, we reviewed the hospital records of 37 children with KD to establish how often the disease manifested "incompletely," and how often pediatricians consulted pediatric ophthalmologists to help in its early diagnosis. Forty-five percent of the children eventually diagnosed with KD lacked the complete diagnostic criteria of KD when admitted to the hospital, and diagnosis and treatment therefore were delayed. Coronary artery aneurysms, a complication that might have less serious consequences if treated earlier, developed in 24% of these patients. Although 67% presented to their pediatrician with bilateral conjunctival injection and/or iridocyclitis, pediatricians requested ophthalmologic consultation for only 5% of them. Our review of these cases indicates that the eye findings in KD could play a role in earlier diagnosis and treatment. Pediatric ophthalmologists and pediatricians should be more aware of their combined responsibilities for expedient recognition of KD.


Aripiprazole, an antipsychotic with a novel mechanism of action, and risperidone vs placebo in patients with schizophrenia and schizoaffective disorder.
Aripiprazole is a dopamine D2 receptor partial agonist with partial agonist activity at serotonin 5HT1A receptors and antagonist activity at 5HT2A receptors. This multicenter trial examined the efficacy, safety, and tolerability of aripiprazole in patients with acute exacerbation of schizophrenia or schizoaffective disorder. In this 4-week double-blind study, 404 patients were randomized to 20 mg/d (n = 101) or 30 mg/d (n = 101) of aripiprazole, placebo (n = 103), or 6 mg/d of risperidone (n = 99). Efficacy assessments included Positive and Negative Syndrome Scale (PANSS) scores and Clinical Global Impression scores. Safety and tolerability evaluations included extrapyramidal symptoms and effects on weight, prolactin, and corrected QT (QTc) interval. Aripiprazole (20 and 30 mg/d) and risperidone (6 mg/d) were significantly better than placebo on all efficacy measures. Separation from placebo occurred at week 1 for PANSS total and positive scores with aripiprazole and risperidone and for PANSS negative scores with aripiprazole. There were no significant differences between aripiprazole and placebo in mean change from baseline in the extrapyramidal symptom rating scales. Mean prolactin levels decreased with aripiprazole but significantly increased 5-fold with risperidone. Mean change in QTc interval did not differ significantly from placebo with any active treatment group. Aripiprazole and risperidone groups showed a similar low incidence of clinically significant weight gain. Aripiprazole is effective, safe, and well tolerated for the positive and negative symptoms in schizophrenia and schizoaffective disorder. It is the first non-D2 receptor antagonist with clear antipsychotic effects and represents a novel treatment development for psychotic disorders.


Conformational and biological properties of glucagon fragments containing residues 1-17 and 19-29.
The 29 amino acid polypeptide hormone glucagon was cleaved into two large fragments by the enzyme clostripain. The conformational properties of these two fragments were monitored by circular dichroism at pH 2 and 12 in both the presence and absence of sodium dodecyl sulfate. Both glucagon (1-17) and glucagon (19-29) have reduced abilities to fold in aqueous solution. However, both fragments can take on structure of higher apparent helical content in acidic solution in the presence of sodium dodecyl sulfate but only the glucagon (19-29) retains this conformation at high pH. Neither of the two fragments react with dimyristoylphosphatidylcholine as the intact peptide does. Only the carboxyl terminal fragment was capable of reacting with an antibody specific for glucagon. The glucagon (1-17) has markedly reduced affinity for binding to the glucagon receptor as well as markedly reduced ability to stimulate adenylate cyclase activity which is not affected by the presence of glucagon (19-29). It is proposed that the intact sequence provides specific groups required for activity as well as the potential for forming a stable amphipathic helix, both of which are necessary for full biological activity at low peptide concentrations.


Use of the internet and self-collected samples as a sexually transmissible infection intervention in rural Illinois communities.
In the USA, reported cases of chlamydia (Chlamydia trachomatis) continue to rise despite substantial funding for screening. National gonorrhoea (Neisseria gonorrhoeae) rates have remained relatively stable, with clusters associated with metropolitan areas. Rural areas are no exception, as every county in Illinois reported cases of chlamydia in 2007. Morbidity associated with infection remains a public health concern, with costs of $US2.5+ billion annually. Novel screening interventions must be examined for their ability to reach those at risk who are missed by traditional methods. The website Iwantthekit.org was modified to allow residents from 25 contiguous counties in Central Illinois to request a self-collected sample kit. Returned kits were tested for chlamydia and gonorrhoea. The initial study period was 12 months. During the study period, 343 kits were requested from 20 counties and 39.9% were satisfactorily returned for analysis. Positivity rates for chlamydia and gonorrhoea were 5.8% and 1.2%, respectively, for females and 1.9% and 0% for males. Males comprised 37.7% of all internet samples (compared with 23.4% for traditional screening venues) and 40.4% of all internet samples submitted by whites (compared with only 17.2% of traditional screening). The female positivity rate was comparable to those seen in other screening venues and the method successfully engaged at-risk males. Overall, participation was low and the costs associated with the program outweighed the averted costs associated with the few cases identified. While this methodology resulted in sample requests from a wide area, it must be utilised by more individuals to become cost-effective.


SMOKING STATUS AND TREATMENT OUTCOMES OF VASCULAR ENDOTHELIAL GROWTH FACTOR INHIBITORS FOR NEOVASCULAR AGE-RELATED MACULAR DEGENERATION.
To assess whether smoking status affects 1-year visual outcomes in eyes treated with vascular endothelial growth factor inhibitors for neovascular age-related macular degeneration. Retrospective analysis of data from a prospectively designed, multicenter, observational database. Nine hundred and eighty seven treatment-naive eyes of patients with neovascular age-related macular degeneration were tracked by the Fight Retinal Blindness! outcome registry in Australia, New Zealand, Singapore, and Switzerland who had documented smoking status at baseline and commenced vascular endothelial growth factor inhibitor therapy from January 2006 to December 2016. Generalized additive models were used to display visual acuity results. There was a significant difference in mean improvement in visual acuity at 12 months between nonsmokers, ex-smokers, and current smokers (7.7 vs. 6.1 vs. 3.5 letters of change; P = 0.046) among patients who completed 12 months of treatment when adjusted for age, baseline visual acuity, and choroidal neovascular membrane lesion type and nested for practice. There was no significant difference in the median number of injections over 12 months of treatment by smoking status. Current smokers were a mean of 6.2 years younger than nonsmokers when they started treatment (P < 0.001). This study found inferior 12-month visual outcomes in patients who continued to smoke while receiving vascular endothelial growth factor inhibitor therapy for neovascular age-related macular degeneration.


Roles for gamma-aminobutyric acid in the development of the paraventricular nucleus of the hypothalamus.
The development of the hypothalamic paraventricular nucleus (PVN) involves several factors that work together to establish a cell group that regulates neuroendocrine functions and behaviors. Several molecular markers were noted within the developing PVN, including estrogen receptors (ER), neuronal nitric oxide synthase (nNOS), and brain-derived neurotrophic factor (BDNF). By contrast, immunoreactive gamma-aminobutyric acid (GABA) was found in cells and fibers surrounding the PVN. Two animal models were used to test the hypothesis that GABA works through GABA(A) and GABA(B) receptors to influence the development of the PVN. Treatment with bicuculline to decrease GABA(A) receptor signaling from embryonic day (E) 10 to E17 resulted in fewer cells containing immunoreactive (ir) ERalpha in the region of the PVN vs. control. GABA(B)R1 receptor subunit knockout mice were used to examine the PVN at P0 without GABA(B) signaling. In female but not male GABA(B)R1 subunit knockout mice, the positions of cells containing ir ERalpha shifted from medial to lateral compared with wild-type controls, whereas the total number of ir ERalpha-containing cells was unchanged. In E17 knockout mice, ir nNOS cells and fibers were spread over a greater area. There was also a significant decrease in ir BDNF in the knockout mice in a region-dependent manner. Changes in cell position and protein expression subsequent to disruption of GABA signaling may be due, in part, to changes in nNOS and BDNF signaling. Based on the current study, the PVN can be added as another site where GABA exerts morphogenetic actions in development.


A brain image database for structure/function analysis.
Lesion-deficit-based structure-function analysis has traditionally been empirical and nonquantitative. Our purpose was to establish a new brain image database (BRAID) that allows the statistical correlation of brain functional measures with anatomic lesions revealed by clinical brain images. Data on 303 participants in the MR Feasibility Study of the Cardiovascular Health Study were tested for lesion/deficit correlations. Functional data were derived from a limited neurologic examination performed at the time of the MR examination. Image data included 3D lesion descriptions derived from the MR examinations by hand segmentation. MR images were normalized in-plane using local, linear Talairach normalization. A database was implemented to support spatial data structures and associated geometric and statistical operations. The database stored the segmented lesions, patient functional scores, and several anatomic atlases. Lesion-deficit association was sought by contingency testing (chi2-test) for every possible combination of each neurologic variable and each labeled atlas structure. Significant associations that confirmed accepted lesion-deficit relationships were sought. Two-hundred thirty-five infarctlike lesions in 117 subjects were viewed collectively after mapping into Talairach cartesian coordinates. Anatomic structures most strongly correlated with neurologic deficits tended to be situated in anatomically appropriate areas. For example, infarctlike lesions associated with visual field defects were correlated with structures in contralateral occipital structures, including the optic radiations and occipital gyri. Known lesion-deficit correlations can be established by a database using a standard coordinate system for representing spatial data and incorporating functional and structural data together with appropriate query mechanisms. Improvements and further applications of this methodology may provide a powerful technique for uncovering new structure-function relationships.


Molecular evidence for the intermolecular S···S interaction in the surface molecular packing motifs of a fused thiophene derivative.
A microscopic investigation of the molecular packing structures of a fused thiophene derivative reveals the important role of intermolecular S···S interaction in directing the 2D self-assembly. Thermal annealing of the assembly results in the irreversible phase transition to a new structure with different molecular trimeric packing motifs.


An optimization-based method for geometrical calibration in cone-beam CT without dedicated phantoms.
In this paper we present a new method for the determination of geometrical misalignments in cone-beam CT scanners, from the analysis of the projection data of a generic object. No a priori knowledge of the object shape and positioning is required. We show that a cost function, which depends on the misalignment parameters, can be defined using the projection data and that such a cost function has a local minimum in correspondence to the actual parameters of the system. Hence, the calibration of the scanner can be carried out by minimizing the cost function using standard optimization techniques. The method is developed for a particular class of 3D object functions, for which the redundancy of the fan beam sinogram in the transaxial midplane can be extended to cone-beam projection data, even at wide cone angles. The method has an approximated validity for objects which do not belong to that class; in that case, a suitable subset of the projection data can be selected in order to compute the cost function. We show by numerical simulations that our method is capable to determine with high accuracy the most critical misalignment parameters of the scanner, i.e., the transversal shift and the skew of the detector. Additionally, the detector slant can be determined. Other parameters such as the detector tilt, the longitudinal shift and the error in the source-detector distance cannot be determined with our method, as the proposed cost function has a very weak dependence on them. However, due to the negligible influence of these latter parameters in the reconstructed image quality, they can be kept fixed at estimated values in both calibration and reconstruction processes without compromising the final result. A trade-off between computational cost and calibration accuracy must be considered when choosing the data subset used for the computation of the cost function. Results on real data of a mouse femur as obtained with a small animal micro-CT are shown as well, proving the capability of the proposed calibration method. In principle, the method can be adapted to other cone-beam imaging modalities (e.g., single photon emission computed tomography).


Immediate versus delayed zoledronic acid for prevention of bone loss in postmenopausal women with breast cancer starting letrozole after tamoxifen-N03CC.
Postmenopausal women with breast cancer (BC) are at increased risk for bone loss. Bisphosphonates improve bone mineral density (BMD) in normal postmenopausal women. The purpose of this study was to determine if immediate treatment with zoledronic acid preserves BMD in postmenopausal women with BC starting letrozole after tamoxifen. Postmenopausal women with BC completing tamoxifen were treated with daily letrozole 2.5 mg/vitamin D 400 I.U., calcium 500 mg twice daily and were randomized to upfront or delayed zoledronic acid 4 mg every 6 months. Patients in the delayed arm were only given zoledronic acid if they developed a post-baseline BMD T score <-2.0 or had a fracture. The primary endpoint was the mean percent change in lumbar spine (LS) BMD at 1 year. About 558 women enrolled; 395 provided 1 year BMD data. The upfront arm experienced a mean change of +3.66% in LS BMD versus -1.66% for the delayed group (P < 0.001). Changes at the femoral neck/total hip were also greater for the upfront versus delayed arms (P < 0.001; P < 0.001) with differences persisting at 2 years. Patients in the delayed arm were more likely to experience a clinically meaningful 5% loss of BMD at all sites versus the upfront zoledronate group. Patients in the upfront arm were slightly more likely to report limb edema, fatigue, fever, nausea and jaw osteonecrosis(1%). Upfront zoledronic acid prevents bone loss in postmenopausal women with BC starting letrozole after tamoxifen.


Antimetabolite pemetrexed primes a favorable tumor microenvironment for immune checkpoint blockade therapy.
The immune checkpoint blockade (ICB) targeting programmed cell death-1 (PD-1) and its ligand (PD-L1) has been proved beneficial for numerous types of cancers, including non-small-cell lung cancer (NSCLC). However, a significant number of patients with NSCLC still fail to respond to ICB due to unfavorable tumor microenvironment. To improve the efficacy, the immune-chemotherapy combination with pemetrexed, cis/carboplatin and pembrolizumab (anti-PD-1) has been recently approved as first-line treatment in advanced NSCLCs. While chemotherapeutic agents exert beneficial effects, the underlying antitumor mechanism(s) remains unclear. Pemetrexed, cisplatin and other chemotherapeutic agents were tested for the potential to induce PD-L1 expression in NSCLC cells by immunoblotting and flow cytometry. The ability to prime the tumor immune microenvironment was then determined by NSCLC/T cell coculture systems and syngeneic mouse models. Subpopulations of NSCLC cells responding differently to pemetrexed were selected and subjected to RNA-sequencing analysis. The key signaling pathways were identified and validated in vitro and in vivo. Pemetrexed induced the transcriptional activation of PD-L1 (encoded by CD274) by inactivating thymidylate synthase (TS) in NSCLC cells and, in turn, activating T-lymphocytes when combined with the anti-PD-1/PD-L1 therapy. Nuclear factor κB (NF-κB) signaling was activated by intracellular reactive oxygen species (ROSs) that were elevated by pemetrexed-mediated TS inactivation. The TS-ROS-NF-κB regulatory axis actively involves in pemetrexed-induced PD-L1 upregulation, whereas when pemetrexed fails to induce PD-L1 expression in NSCLC cells, NF-κB signaling is unregulated. In syngeneic mouse models, the combinatory treatment of pemetrexed with anti-PD-L1 antibody created a more favorable tumor microenvironment for the inhibition of tumor growth. Our findings reveal novel mechanisms showing that pemetrexed upregulates PD-L1 expression and primes a favorable microenvironment for ICB, which provides a mechanistic basis for the combinatory chemoimmunotherapy in NSCLC treatment.


The use of inductively coupled plasma mass spectrometry as a detector in drug metabolism studies.
The inherent properties of element selectivity combined with high sensitivity and structure independent response, make inductively coupled plasma mass spectrometry (ICP-MS) an interesting alternative detection technique in drug metabolism studies. The application of online separation with ICP-MS detection in drug metabolism studies is reviewed with focus on the merits and demerits of this detection technique. The prerequisite for inclusion in this review is that the study involves a separation technique hyphenated online to the ICP-MS detection. ICP-MS detection is found to be advantageous for analysis of all drug substances detectable by ICP-MS compared to radiochemical detection. Detectable drugs are limited to halogen-, sulfur-, metal- and metalloid-containing compounds. The drawback of interference from endogenous compounds on quantitative mass balance estimations of non-metal drugs is addressed. The potential of determining the stoichiometry in metallo-drug biomolecule interactions is pointed out by presenting examples of simultaneous monitoring of metals in metallo-drugs and intrinsic ICP-MS detectable elements in biomolecules. It is concluded that ICP-MS detection is an indispensable technique in drug metabolism studies of metallo-drugs, although the applicability for traditional drugs is limited.


Emerging roles for the influenza A virus nuclear export protein (NEP).
Influenza virus is a major human and animal pathogen causing seasonal epidemics and occasional pandemics in the human population that are associated with significant morbidity and mortality. Influenza A virus, a member of the orthomyxovirus family, contains an RNA genome with a coding capacity for a limited number of proteins. In addition to ensuring the structural integrity of virions, these viral proteins facilitate the replication of virus in the host cell. Consequently, viral proteins often evolve to perform multiple functions, the influenza A virus nuclear export protein (NEP) (also referred to as non-structural protein 2, or NS2) being an emerging example. NEP was originally implicated in mediating the nuclear export of viral ribonucleoprotein (RNP) complexes, which are synthesized in the infected cell nucleus and are assembled into progeny virions at the cell membrane. However, since then, new and unexpected roles for NEP during the influenza virus life cycle have started to emerge. These recent studies have shown NEP to be involved in regulating the accumulation of viral genomic vRNA and antigenomic cRNA as well as viral mRNA synthesized by the viral RNA-dependent RNA polymerase. Subsequently, this regulation of viral RNA transcription and replication by NEP was shown to be an important factor in the adaptation of highly pathogenic avian H5N1 influenza viruses to the mammalian host. Unexpectedly, NEP has also been implicated in recruiting a cellular ATPase to the cell membrane to aid the efficient release of budding virions. Accordingly, NEP is proposed to play multiple biologically important roles during the influenza virus life cycle.


[The placental histo- and ultrastructure peculiarities during the cytomegalovirus infection in pregnant].
Object of the investigation - estimation of changes of the feto-placental complex and syncytiotrophoblaston based on the placental morphology and the activity of the antiviral antibodies in expectant mothers. The work is based on findings from 50 expectant mothers with cytomegalovirus (CMV) infection - 43 with reactivation of the latent infection according to the data of high avid IgG (86%) and 7 with primary active CMV infection according the level of the low avid IgG (14%). 20 healthy expectant mothers were included in the control group. It was shown that the active primary and reactivated CMV infection accompanied by the drawing in pathologic process the chorion structures in a productive inflammation - chorionamnionis and choriovillusitis manner, which testify to drawing the fetus immune system in reciprocal reaction with subsequent intrauterine growth retardation.


Global prevalence, cascade of care, and prophylaxis coverage of hepatitis B in 2022: a modelling study.
The 2016 World Health Assembly endorsed the elimination of hepatitis B virus (HBV) infection as a public health threat by 2030; existing therapies and prophylaxis measures make such elimination feasible, even in the absence of a virological cure. We aimed to estimate the national, regional, and global prevalence of HBV in the general population and among children aged 5 years and younger, as well as the rates of diagnosis, treatment, prophylaxis, and the future burden globally. In this modelling study, we used a Delphi process with data from literature reviews and interviews with country experts to quantify the prevalence, diagnosis, treatment, and prevention measures for HBV infection. The PRoGReSs Model, a dynamic Markov model, was used to estimate the country, regional, and global prevalence of HBV infection in 2022, and the effects of treatment and prevention on disease burden. The future incidence of morbidity and mortality in the absence of additional interventions was also estimated at the global level. We developed models for 170 countries which resulted in an estimated global prevalence of HBV infection in 2022 of 3·2% (95% uncertainty interval 2·7-4·0), corresponding to 257·5 million (216·6-316·4) individuals positive for HBsAg. Of these individuals, 36·0 million were diagnosed, and only 6·8 million of the estimated 83·3 million eligible for treatment were on treatment. The prevalence among children aged 5 years or younger was estimated to be 0·7% (0·6-1·0), corresponding to 5·6 million (4·5-7·8) children with HBV infection. Based on the most recent data, 85% of infants received three-dose HBV vaccination before 1 year of age, 46% had received a timely birth dose of vaccine, and 14% received hepatitis B immunoglobulin along with the full vaccination regimen. 3% of mothers with a high HBV viral load received antiviral treatment to reduce mother-to-child transmission. As 2030 approaches, the elimination targets remain out of reach for many countries under the current frameworks. Although prevention measures have had the most success, there is a need to increase these efforts and to increase diagnosis and treatment to work towards the elimination goals. John C Martin Foundation, Gilead Sciences, and EndHep2030.


[Influence of infection related factors on neuropsychological performance of HIV seropositive subjects].
There are no definitive risk factors which predict the development of neuropsychological deficits associated to HIV infection, and, taking into account that neuropsychological impairment is a risk factor of early death in all stages of HIV infection, there is a pressing need to determine those factors, either related or foreign to HIV, which help to explain the existing intragroup differences and that may also be associated with the development and progression of the dysfunction. The aim of this work is to revise those factors related with the infection that can influence neuropsychological functioning of the seropositive patients, such as antiretroviral treatments, immunological impairment, viral load and the advance of the infection. Identification of these risk factors will help us to outline characteristics of the seropositive subjects who are neuropsychologically more vulnerable, in order to detect them early with a view to preventing the manifestation of neuropsychological disturbances, and offer quick, suitable treatment.


Telemental Health Collaborative Care Medication Management: Implementation and Outcomes.
Introduction: Access to quality mental health medication management (MM) in the United States is limited, even among those with employment-based health insurance. This implementation, feasibility, and outcome study sought to design and evaluate an evidence-based telemental health MM service using a collaborative care model (CoCM). Materials and Methods: CoCM MM was available to adult employees/dependents through their employer benefits, in addition to therapy. Outcomes included Patient Health Questionnaire-9 (PHQ-9) and the Generalized Anxiety Disorder-7 (GAD-7) collected at baseline and throughout participation. This analysis was not deemed to be human subjects research by the Western Institutional Review Board. Results: Over 17 months, 212 people enrolled and completed >2 assessments; the enrollees were 58.96% female with average age of 32.00 years (standard deviation [SD] = 7.38). In people with moderate to severe depression or anxiety, PHQ-9 and GAD-7 scores reduced by an average of 7.27 (SD = 4.80) and 6.71 (SD = 5.18) points after at least 12 ± 4 weeks in the program. At 24 ± 4 weeks, the PHQ-9 and GAD-7 reductions were on average 7.17 (SD = 5.00) and 6.03 (SD = 5.37), respectively. Approximately 65.88% of participants with either baseline depression or anxiety had a response on either the PHQ-9 or GAD-7 at 12 ± 4 weeks and 44.71% of participants experienced remission; at 24 ± 4 weeks, 56.41% had response and 41.03% experienced remission. Conclusions: An evidence-based CoCM telemedicine service within an employee behavioral health benefit is feasible and effective in reducing anxiety and depression symptoms when using measurement-based care. Widespread implementation of a benefit like this could expand access to evidence-based mental health MM.


Social class, smoking and the severity of respiratory symptoms in the general population.
The prevalence of respiratory symptoms has been found in some studies to vary with social class. One explanation of this link may be the effect of exposure to cigarette smoke. To investigate this, the relation between social class, smoking and respiratory symptoms was explored in a population based survey. A cross sectional survey using a validated questionnaire. Two general practices in Staffordshire, United Kingdom. A random sample of 4237 patients aged 16 and over from two general practices in Staffordshire were mailed a questionnaire enquiring about respiratory symptoms and their severity. The severity of respiratory symptoms increased with increasing exposure to cigarette smoke and was greater among manual social classes. Current smokers (odds ratio (OR) = 2.9, 95% confidence limits (CI) 2.3, 3.6), past smokers (OR = 1.5, 95% CI 1.2, 1.8) and passive smokers (OR = 1.4, 95% CI 1.0, 1.8) were more likely to report the more severe respiratory symptoms compared with non-smokers. Responders from social class V (OR = 2.4, 95% CI 1.3, 4. 4) were more likely to report the more severe respiratory symptoms compared with social class I, as were responders from social classes IIIM (OR = 1.3, 95% CI 0.9, 1.9) and IV (OR = 1.4, 95% CI 0.9, 2.1). These effects were independent of each other. This study has shown that social class is linked to the severity of respiratory symptoms, independently of smoking. Although the need to reduce and quit smoking in manual class households remains a crucial preventive issue, other mechanisms by which social class differences may influence symptom occurrence and severity need to be explored.


Removal of amoxicillin and cefuroxime axetil by advanced membranes technology, activated carbon and micelle-clay complex.
Two antibacterials, amoxicillin trihydrate and cefuroxime axetil spiked into wastewater were completely removed by sequential wastewater treatment plant's membranes, which included activated sludge, ultrafiltration (hollow fibre and spiral wound membranes with 100 and 20 kDa cut-offs), activated carbon column and reverse osmosis. Adsorption isotherms in synthetic water which employed activated carbon and micelle-clay complex (octadecyltrimethylammonium-montmorillonite) as adsorbents fitted the Langmuir equation. Qmax of 100 and 90.9 mg g(-1), and K values of 0.158 and 0.229 L mg(-1) were obtained for amoxicillin trihydrate using activated carbon and micelle-clay complex, respectively. Filtration of antibacterials in the ppm range, which yielded variable degrees of removal depending on the volumes passed and flow rates, was simulated and capacities for the ppb range were estimated. Stability study in pure water and wastewater revealed that amoxicillin was totally stable for one month when kept at 37°C, whereas cefuroxime axetil underwent slow hydrolysis to cefuroxime.


[Posplacenta intrauterine device. Guide wires not visible].
Little information exists about the follow- up of the strings of intrauterine device (IUD) applied posplacenta. When the strings are of 10 cm in length are within the uterine cavity should be visible after postpartum involution. To know the incidence of visible strings of intrauterine device (IUD) after posplacental insertion and to establish guidelines of follow-up when strings are not seen. A prospective cohort study of 530 women with posplacental insertion of T Cu 380-A IUD with strings 10 cm long. Follow-up was at 6 weeks, 3, 6, 9 and 12 months after IUD insertion, at each visit women were questioned about IUD expulsion or removal and the cervix was inspected to visualize the strings of IUD. We analyzed the cumulative incidence of visible strings and procedures that were performed to locate the IUD when strings were not seen. At one year of follow-up the IUD was in situ in 398 women (75.1%), there were 44 expulsions (8.3%), 24 removals (4.5%) and 64 lost to follow up (12.1%). In women with IUDs in situ the strings were seen in 90.2% (n = 359) and were not seen in 9.8% (n = 39). In 65 women uterine cavity was probed identifying the IUD in situ in 64 cases, we performed 33 pelvic ultrasounds with IUDs in situ in 31 cases and in 2 cases with IUD in the cervical canal.. Strings of IUD after postplacental insertion was seen in most women at one year of follow up. When we are reasonably sure the woman is not pregnant, interventions for the location of the IUD should be avoided. A pelvic ultrasound should be the initial procedure and usually sufficient to locate IUDs whose strings are not visible.


Ca:Mg Ratio, APOE Cytosine Modifications, and Cognitive Function: Results from a Randomized Trial.
Deterioration of ionized calcium (Ca2+) handling in neurons could lead to neurodegenerative disease. Magnesium (Mg) antagonizes Ca during many physiologic activities, including energy metabolism and catalyzation of demethylation from 5-methylcytosine(5-mC) to 5-hydroxymethylcytosine(5-hmC). To test the hypothesis that actively reducing the Ca:Mg intake ratio in the diet through Mg supplementation improves cognitive function, and to test whether this effect is partially mediated by modified cytosines in Apolipoprotein E (APOE). This study is nested within the Personalized Prevention of Colorectal Cancer Trial (PPCCT), a double-blind 2×2 factorial randomized controlled trial, which enrolled 250 participants from Vanderbilt University Medical Center. Target doses for both Mg and placebo arms were personalized. Among those aged > 65 years old who consumed a high Ca:Mg ratio diet, we found that reducing the Ca:Mg ratio to around 2.3 by personalized Mg supplementation significantly improved cognitive function by 9.1% (p = 0.03). We also found that reducing the Ca:Mg ratio significantly reduced 5-mC at the cg13496662 and cg06750524 sites only among those aged > 65 years old (p values = 0.02 and 0.03, respectively). Furthermore, the beneficial effect of reducing the Ca:Mg ratio on cognitive function in those aged over 65 years was partially mediated by reductions in 5-mC levels (i.e., cg13496662 and cg06750524) in APOE (p for indirect effect = 0.05). Our findings suggest that, among those age 65 and over with a high dietary Ca:Mg ratio, optimal Mg status may improve cognitive function partially through modifications in APOE methylation. These findings, if confirmed, have significant implications for the prevention of cognitive aging and Alzheimer's disease.Clinical Trial Registry number and website: #100106 https://clinicaltrials.gov/ct2/show/NCT03265483.


Multiparametric PET/CT-perfusion does not add significant additional information for initial staging in lung cancer compared with standard PET/CT.
The purpose of this study was to assess the relationship of CT-perfusion (CTP), 18F-FDG-PET/CT and histological parameters, and the possible added value of CTP to FDG-PET/CT in the initial staging of lung cancer. Fifty-four consecutive patients (median age 65 years, 15 females, 39 males) with suspected lung cancer were evaluated prospectively by CT-perfusion scan and 18F-FDG-PET/CT scan. Overall, 46 tumors were identified. CTP parameters blood flow (BF), blood volume (BV), and mean transit time (MTT) of the tumor tissue were calculated. Intratumoral microvessel density (MVD) was assessed quantitatively. Differences in CTP parameters concerning tumor type, location, PET positivity of lymph nodes, TNM status, and UICC stage were analyzed. Spearman correlation analyses between CTP and 18F-FDG-PET/CT parameters (SUVmax, SUVmean, PETvol, and TLG), MVD, tumor size, and tumor stage were performed. The mean BF (mL/100 mL min-1), BV (mL/100 mL), and MTT (s) was 35.5, 8.4, and 14.2, respectively. The BF and BV were lower in tumors with PET-positive lymph nodes (p = 0.02). However, the CTP values were not significantly different among the N stages. The CTP values were not different, depending on tumor size and location. No significant correlation was found between CTP parameters and MVD. Overall, the CTP information showed only little additional information for the initial staging compared with standard FDG-PET/CT. Low perfusion in lung tumors might possibly be associated with metabolically active regional lymph nodes. Apart from that, both CTP and 18F-FDG-PET/CT parameter sets may reflect different pathophysiological mechanisms in lung cancer.


Immunopathogenesis Versus Protection in Dengue Virus Infections.
Dengue viruses (DENV) are mosquito-borne viruses that cause significant morbidity. The existence of four serotypes of DENV with partial immunologic cross-reactivity creates the opportunity for individuals to experience multiple acute DENV infections over the course of their lifetimes. Research over the past several years has revealed complex interactions between DENV and the human innate and adaptive immune systems that can have either beneficial or detrimental influences on the outcome of infection. Further studies that seek to distinguish protective from pathological immune responses in the context of natural DENV infection as well as clinical trials of candidate DENV vaccines have an important place in efforts to control the global impact of this re-emerging viral disease.


The effect of extended calving intervals in high-yielding lactating cows on milk production and profitability.
A field trial was conducted to examine the effect of extended calving interval (CI) on production and profitability of high yielding cows (n = 937). First insemination was performed at 154 and 93 d postpartum (pp), for treatment and control primiparous cows, respectively, and at 124 and 71 d pp for treatment and control multiparous cows, respectively. During the first experimental lactation, average daily value-corrected milk (VCM) yield was 28.5 and 27.7 kg/d of CI for treatment (n = 131) and control (n = 133) primiparous cows, respectively. No significant difference in average daily VCM yield (33.0 and 32.8 kg/d of CI) was found between treatment (n = 271) and control (n = 215) multiparous cows . In the first 150 d of the subsequent lactation, there were significant differences in milk and VCM production in favor of the treatment primiparous cows (41.4 vs. 39.7 kg of VCM/d) but no significant differences in the production of multiparous cows. Primiparous and multiparous cows with extended lactations were more profitable. During the first experimental lactation, there were advantages of $0.19 and $0.12/d of CI in the net returns for primiparous cows and multiparous cows with longer CI, respectively. When the economic analysis included the first experimental lactation plus the first 150 d of the subsequent one, the net return per day of CI was higher for cows with an extended voluntary waiting period: $0.21/d and $0.16/d for primiparous and multiparous cows, respectively. A delay of 60 d with respect to the usual voluntary waiting period in the beginning of inseminations of high yielding cows has economic advantages and allows the farmer an option for decisions regarding individual cows.


Flying with the wind: scale dependency of speed and direction measurements in modelling wind support in avian flight.
Understanding how environmental conditions, especially wind, influence birds' flight speeds is a prerequisite for understanding many important aspects of bird flight, including optimal migration strategies, navigation, and compensation for wind drift. Recent developments in tracking technology and the increased availability of data on large-scale weather patterns have made it possible to use path annotation to link the location of animals to environmental conditions such as wind speed and direction. However, there are various measures available for describing not only wind conditions but also the bird's flight direction and ground speed, and it is unclear which is best for determining the amount of wind support (the length of the wind vector in a bird's flight direction) and the influence of cross-winds (the length of the wind vector perpendicular to a bird's direction) throughout a bird's journey. We compared relationships between cross-wind, wind support and bird movements, using path annotation derived from two different global weather reanalysis datasets and three different measures of direction and speed calculation for 288 individuals of nine bird species. Wind was a strong predictor of bird ground speed, explaining 10-66% of the variance, depending on species. Models using data from different weather sources gave qualitatively similar results; however, determining flight direction and speed from successive locations, even at short (15 min intervals), was inferior to using instantaneous GPS-based measures of speed and direction. Use of successive location data significantly underestimated the birds' ground and airspeed, and also resulted in mistaken associations between cross-winds, wind support, and their interactive effects, in relation to the birds' onward flight. Wind has strong effects on bird flight, and combining GPS technology with path annotation of weather variables allows us to quantify these effects for understanding flight behaviour. The potentially strong influence of scaling effects must be considered and implemented in developing sampling regimes and data analysis.


[Protective effects of cilostazol against transient focal cerebral ischemia and hemorrhagic transformation].
Cilostazol, a selective inhibitor of phosphodiesterase III, is an antiplatelet drug and a vasodilator via increased cAMP levels. It has been approved for the treatment of ischemic symptoms in chronic peripheral arterial obstruction or intermittent claudication and for secondary prevention of cerebral infarction (CSPS I). Recently, cilostazol has been reported to be more effective than aspirin in the secondary prevention of all types of stroke in patients and, in particular, prevent the secondary attack of hemorrhagic stroke in patients (CSPS II). Laboratory investigations revealed that cilostazol has a neuroprotective effect against ischemic brain injury. The neuroprotective potential is dependent on its antiinflammatory and antiapoptotic effects mediated by scavenging hydroxyl radicals, decreasing formation of tumor necrosis factor-α, and inhibition of poly (ADP-ribose) polymerase activity. In addition, increasing evidence indicates that cilostazol may offer endothelial protection via both the inhibition of lipopolysaccharide-induced apoptosis and induced nitric oxide (NO) production by endothelial NO synthase activation. The breakdown of the barrier permeability of the blood brain barrier (BBB) often accelerates the progression of diseases such as cerebral ischemia. However, the molecular mechanisms involved in BBB disruption have not been fully determined. Identification of the molecules responsible for the disruption of the endothelial barrier may yield new therapeutic targets in intractable diseases. This article reviews the protective effects of cilostazol against transient focal cerebral ischemia and hemorrhagic transformation and its mechanism of action.


The genetic basis of a rare flower color polymorphism in Mimulus lewisii provides insight into the repeatability of evolution.
A long-standing question in evolutionary biology asks whether the genetic changes contributing to phenotypic evolution are predictable. Here, we identify a genetic change associated with segregating variation in flower color within a population of Mimulus lewisii. To determine whether these types of changes are predictable, we combined this information with data from other species to investigate whether the spectrum of mutations affecting flower color transitions differs based on the evolutionary time-scale since divergence. We used classic genetic techniques, along with gene expression and population genetic approaches, to identify the putative, loss-of-function mutation that generates rare, white flowers instead of the common, pink color in M. lewisii. We found that a frameshift mutation in an anthocyanin pathway gene is responsible for the white-flowered polymorphism found in this population of M. lewisii. Comparison of our results with data from other species reveals a broader spectrum of flower color mutations segregating within populations relative to those that fix between populations. These results suggest that the genetic basis of fixed differences in flower color may be predictable, but that for segregating variation is not.


[Antibacterial action of ether oils of some plants].
Inhibitory effect of clove oil on Escherichia coli, Staphylococcus aureus, Salmonella typhimurium, Shigella dysenteriae and Candida albicans was detected. Mint ether oil had the high antibacterial action on S. aureus, however against other microorganisms mint oil had a reliably low effect then clove oil. Fennel oil had high antibacterial effect on C. albicans, and bactericidal action on S. typhimurium and S. dysenteriae.


Prenatal drug exposure and maternal and infant feeding behaviour.
To evaluate feeding difficulties and maternal behaviour during a feeding session with 1 month old infants prenatally exposed to cocaine and/or opiates. The study is part of the maternal lifestyle study, which recruited 11 811 subjects at four urban hospitals, then followed 1388 from 1 to 36 months of age. Exposure to cocaine and opiates was determined by maternal interview and meconium assay. At the 1 month clinic visit, biological mothers were videotaped while bottle feeding their infants. This sample included 364 exposed to cocaine, 45 exposed to opiates, 31 exposed to both drugs, and 588 matched comparison infants. Mothers were mostly black, high school educated, and on public assistance. Videotapes were coded without knowledge of exposure status for frequency, duration and quality of infant sucking, arousal, feeding problems, and maternal feeding activity and interaction. No cocaine effects were found on infant feeding measures, but cocaine-using mothers were less flexible (6.29 v 6.50), less engaged (5.77 v 6.22), and had shorter feeding sessions (638 v 683 seconds). Opiate exposed infants showed prolonged sucking bursts (29 v 20 seconds), fewer pauses (1.6 v 2.2 per minute), more feeding problems (0.55 v 0.38), and increased arousal (2.59 v 2.39). Their mothers showed increased activity (30 v 22), independent of their infants' feeding problems. Previous concerns about feeding behaviour in cocaine exposed infants may reflect the quality of the feeding interaction rather than infant feeding problems related to prenatal exposure. However, opiate exposed infants and their mothers both contributed to increased arousal and heightened feeding behaviour.


H2-rich fluids from serpentinization: geochemical and biotic implications.
Metamorphic hydration and oxidation of ultramafic rocks produces serpentinites, composed of serpentine group minerals and varying amounts of brucite, magnetite, and/or FeNi alloys. These minerals buffer metamorphic fluids to extremely reducing conditions that are capable of producing hydrogen gas. Awaruite, FeNi3, forms early in this process when the serpentinite minerals are Fe-rich. Olivine with the current mantle Fe/Mg ratio was oxidized during serpentinization after the Moon-forming impact. This process formed some of the ferric iron in the Earth's mantle. For the rest of Earth's history, serpentinites covered only a small fraction of the Earth's surface but were an important prebiotic and biotic environment. Extant methanogens react H2 with CO2 to form methane. This is a likely habitable environment on large silicate planets. The catalytic properties of FeNi3 allow complex organic compounds to form within serpentinite and, when mixed with atmospherically produced complex organic matter and waters that circulated through basalts, constitutes an attractive prebiotic substrate. Conversely, inorganic catalysis of methane by FeNi3 competes with nascent and extant life.


Recommandations of the Tunisian Association of Surgery for the practice of visceral surgery during COVID-19 pandemic.
The World Health Organization declared on March 11, 2020 that the COVID-19 epidemic has become a pandemic. In Tunisia, the Ministry of Health has recommended enhanced preventive hygiene measures to contain and limit the spread of the virus. Following the entry of Tunisia into phase 4 of the COVID-19 epidemic, the Tunisian Association of Surgery proposed recommendations related to surgical activity. Surgical emergencies must be treated urgently and without delay. Non-tumor pathologies which require surgery in an elective situation and for which the risk of aggravation or complication is considered low shoud be postponed. For digestive tumor pathology, and apart from complicated forms, neoadjuvant treatment is highly recommended in the context of multidisciplinary concertation staff.


Evaluation and application of a luciferase fusion system for rapid in vivo analysis of RNAi targets and constructs in plants.
The practical use of RNA-mediated approaches including antisense RNA, ribozymes and siRNAs for specific inhibition of gene expression is limited by lack of simple quantitative methods to rapidly test efficacy in vivo. There have been indications that cotransfer of target::reporter gene fusions with constructs designed against the target sequence, followed by quantification of transient reporter gene activity might be effective. Here, we report detailed testing of the approach in plants, using diverse target::luciferase fusions and antisense or ribozyme constructs. We used quantitative transient luciferase activity (Luc) assays to test antisense constructs against beta-glucuronidase, PR glucanase, vacuolar invertase and cucumber mosaic virus, as well as ribozymes against watermelon mosaic virus and tobacco anionic peroxidase. For constructs previously tested in transgenic plants, the results correspond well with those from the transient expression assay. Target susceptibility was generally not strongly influenced by luciferase fusion, and the assay was not highly dependent on target sequence length. Some sequences reduced Luc activity below the level for reliable quantification, but suitable alternative fusions were readily produced. Transcriptional and translation fusions were effective for 5' target::luc constructs. Translational fusions were more reliable for luc::target 3' constructs. With minimal preliminary work to prepare suitable target::luciferase fusions, the approach appears generally applicable for rapid in vivo validation of effectiveness and specificity of constructs designed for RNA-mediated down-regulation of plant genes.


Metabolically obese normal-weight children.
Non-obese children with elevated serum insulin levels and metabolic disorders such as, hyperglycemia, hypertension, and/or hypertriglyceridemia are a subset of children in high risk of developing cardiovascular disease later in life. Since usually the health policies for the prevention of the obesity associated disorders in children are based on the screening focused on the obese, frequently the metabolically obese normal-weight (MONW) children are not identified in primary care setting. Given that characterization of the MONW children is an important public health issue, and that a large amount of resources might be unnecessarily used in the screening of metabolic risk of nonobese children; we review data regarding criteria for the early recognition of this subset of children in high risk of developing cardiovascular disease. Results of our review suggests that the presence of family history of type 2 diabetes and/or hypertension, the elevated percentage of body fat, and the high birth-weight should be taken into account as criteria of high cardiovascular risk, irrespective of obesity.


Nano-Plumber Reshapes Glymphatic-Lymphatic System to Sustain Microenvironment Homeostasis and Improve Long-Term Prognosis after Traumatic Brain Injury.
Traumatic brain injury (TBI) is a leading cause of death and disability worldwide. Long-term changes in the microenvironment of the brain contribute to the degeneration of neurological function following TBI. However, current research focuses primarily on short-term modulation during the early phases of TBI, not on the critical significance of long-term homeostasis in the brain microenvironment. Notably, dysfunction of the glymphatic-lymphatic system results in the accumulation of danger/damage-associated molecular patterns (DAMPs) in the brain, which is regarded as the leading cause of long-term microenvironmental disturbances following TBI. Here, a nanostructure, Nano-plumber, that co-encapsulates the microenvironment regulator pro-DHA and the lymphatic-specific growth factor VEGF-C is developed, allowing for a sustainable and orderly regulation of the microenvironment to promote long-term neurological recovery. Nano-plumber reverses the injury microenvironment by suppressing microglia and astrocytes activation and maintaining reduced activation via enhanced glymphatic-lymphatic drainage, and significantly improves the neurological function of rodents with TBI. This study demonstrates that glymphatic-lymphatic system reconstruction is essential for enhancing long-term prognosis following TBI, and that the Nano-plumber developed here may serve as a clinically translatable treatment option for TBI.


Effects of whole-body vibration with an unstable surface on muscle activation.
The current study examined the effects of using an unstable surface during whole-body vibration (WBV) exercise on leg and trunk muscle activity during a static semi-squat. Twenty-eight recreationally active university students completed 4 different test conditions: 1) stable surface with no WBV; 2) unstable surface with no WBV; 3) unstable surface with 30 Hz WBV low amplitude; and 4) unstable surface with 50 Hz WBV low amplitude. Surface electromyography (sEMG) was measured for the gastrocnemius medialis (GM), vastus medialis oblique (VMO), vastus lateralis (VL), rectus abdominis (RA), and multifidus (MF) muscles. Normalized to the stable condition, WBV at 30 Hz and an unstable surface increased EMG in the GM vs the unstable and stable surfaces (~35%; p<0.05). VMO EMG decreased in the unstable vs stable condition (~20%), WBV at 30 Hz and an unstable surface increased EMG vs all other conditions (~20-40%; p<0.05). MF EMG increased with WBV at 30 Hz (25%; p<0.05) vs the stable condition but not vs all other conditions. Using an unstable surface during WBV exposure increases EMG of muscles in the lower extremities and trunk suggesting the combination of an unstable surface combined with WBV may be an effective modality to further increase EMG.


Quantitative Lys-ϵ-Gly-Gly (diGly) proteomics coupled with inducible RNAi reveals ubiquitin-mediated proteolysis of DNA damage-inducible transcript 4 (DDIT4) by the E3 ligase HUWE1.
Targeted degradation of proteins through the ubiquitin-proteasome system (UPS) via the activities of E3 ubiquitin ligases regulates diverse cellular processes, and misregulation of these enzymes contributes to the pathogenesis of human diseases. One of the challenges facing the UPS field is to delineate the complete cohort of substrates for a particular E3 ligase. Advances in mass spectrometry and the development of antibodies recognizing the Lys-ϵ-Gly-Gly (diGly) remnant from ubiquitinated proteins following trypsinolysis have provided a tool to address this question. We implemented an inducible loss of function approach in combination with quantitative diGly proteomics to find novel substrates of HUWE1 (HECT, UBA, and WWE domain containing 1, E3 ubiquitin protein ligase), an E3 ligase implicated in cancer and intellectual disabilities. diGly proteomics results led to the identification of DNA damage-inducible transcript 4 (DDIT4) as a putative HUWE1 substrate. Cell-based assays demonstrated that HUWE1 interacts with and regulates ubiquitination and stability of DDIT4. Together these data suggest a model in which HUWE1 mediates DDIT4 proteasomal degradation. Our results demonstrate proof of concept that inducible knockdown of an E3 ligase in combination with diGly proteomics provides a potentially advantageous method for identifying novel E3 substrates that may help to identify candidates for therapeutic modulation in the UPS.


Menopausal Symptoms according to Sasang Constitution and Menopausal Status among Perimenopause Women.
The purpose of this study was to address the difference menopausal symptoms according to sasang constitution and menopausal status among perimenopausal women. Data for this cross-sectional study was collected by administering questionnaires that elicited general information. The menopause rating scale (MRS) and Questionnaire for the Sasang Constitution Classification II (QSCC II) was given to 284 perimenopausal women who met the eligibility criteria and agreed to participate in the study. The study subjects were composed of 27.5% so-yangin, 24.6% so-eumin, 18.7% tae-eumin, and 29.6% undefined category. The total MRS and all subscales were significantly higher for postmenopausal women in comparison to premenopausal women. Especially, so-eumin and tae-eumin displayed significantly higher scores in urogenital symptoms. The MRS score in postmenopausal women who belonged to the group of tae-eumin and so-eumin was significantly higher than those for premenopausal women. However, so-yangin displayed higher scores in psychological and urogenital symptoms than tae-eumin for premenopausal women. So-eumin presented significantly higher scores in urogenital symptoms in comparison to the undefined category for postmenopausal women. These findings suggest that sasang constitution could be an important factor in understanding the woman's menopausal symptoms and identify the best treatment.


Women's perspectives about cannabis use during pregnancy and the postpartum period: An integrative review.
Cannabis is the most commonly used illicit drug among general and pregnant populations. Despite recommendations to abstain from cannabis use, its use is increasing during the perinatal period. In this integrative review, we aim to understand women's perspectives about the health aspects of perinatal cannabis use. The following databases were searched: MEDLINE, PsycInfo, EMBASE, and CINAHL. We included quantitative and qualitative studies with a primary focus on perinatal cannabis use. The methodological quality of the included studies was appraised using appropriate checklists. Extracted data were integrated into a single data matrix and iteratively compared across studies to summarize and synthesize the data. Full text of 33 citations was retrieved and reviewed of which 6 studies met the inclusion criteria. Women who continued to use cannabis during pregnancy often perceived no general or pregnancy-specific risk compared to nonusers. The uncertainty regarding adverse perinatal consequences, its perceived therapeutic effects, and lower costs of cannabis compared to that of cigarettes contributed to cannabis use. A lack of communication with health care providers regarding the health aspects of cannabis was evident. Women perceived this lack of counseling as an indication that adverse outcomes associated with cannabis use are not significant. Women's perceptions of health risks associated with cannabis use are important factors in the decision-making process regarding use or cessation, particularly as legal concerns are fading away. A discussion about health concerns surrounding cannabis use may influence women's perceptions of risk and help them to make informed choices.


[Normoaldosterone spironolactone sensitive hypertension].
We report the case of normoaldosterone spironolactone sensitive hypertension. Patients suffering from this recently identified form of arterial hypertension exhibit clinical symptoms resembling the symptoms of primary aldosteronism. The described disturbances may be misdiagnosed as essential hypertension because plasma aldosterone levels and aldosterone/renin ratio in patients are within the normal ranges. In the described state of the examined female, detection of the disease and implementation of spironolactone treatment not only normalised arterial blood pressure but also allowed to avoid unnecessary administration of many antihypertensive agents.