Possible sources of glutamine for parenteral nutrition: impact on glutamine metabolism.
Due to its instability, glutamine is not included in solutions for parenteral solution. This problem can be obviated by providing glutamine as acetyl-, glycyl-, or alanylglutamine. Using an organ balance technique in conscious dogs, we investigated metabolism of these three sources of glutamine. Liver, gut, kidney, and muscle participated in clearance of glycyl- and alanylglutamine from plasma, but among these organs only kidney cleared acetylglutamine. Furthermore, there was a large urinary excretion for acetylglutamine (38 +/- 6% of amount infused) but only a trace amount for either dipeptide. The infusion of glutamine-dipeptides resulted in similar increases in blood level of free glutamine. The main source of this increase appeared to be hydrolysis of dipeptides by kidney and release of free glutamine to circulation. During the infusion of both dipeptides, glutamine balance (free and dipeptide forms) was always positive (net uptake) across liver, gut, and kidney but was neutral across muscle. Liver or gut glutamine balances were not significantly different during the infusion of dipeptides, but kidney glutamine balance was twofold greater during the infusion of glycyl- than alanylglutamine. We conclude that among these three sources of glutamine, acetylglutamine is least desirable for use in parenteral nutrition. Glycylglutamine may be preferable over alanylglutamine if the objective is to target glutamine for kidney.


Construction of self-cloning, indigenous wine strains of Saccharomyces cerevisiae with enhanced glycerol and glutathione production.
To improve wine taste and flavor stability, a novel indigenous strain of Saccharomyces cerevisiae with enhanced glycerol and glutathione (GSH) production for winemaking was constructed. ALD6 encoding an aldehyde dehydrogenases of the indigenous yeast was replaced by a GPD1 and CUP1 gene cassette, which are responsible for NAD-dependent glycerol-3-phosphatase dehydrogenase and copper resistance, respectively. Furthermore, the α-acetohydroxyacid synthase gene ILV2 of the indigenous yeast was disrupted by integration of the GSH1 gene which encodes γ-glutamylcysteine synthetase and the CUP1 gene cassette. The fermentation capacity of the recombinant was similar to that of the wild-type strain, with an increase of 21 and 19 % in glycerol and GSH production. No heterologous DNA was harbored in the recombinant in this study.


A method for timely assessment of influenza-associated mortality in the United States.
Influenza-associated mortality has traditionally been estimated as the excess mortality above a baseline of deaths during influenza epidemic periods. Excess mortality estimates are not timely, because national vital statistics data become available after a period of 2-3 years. To develop a method for timely reporting, we used the 121 Cities Surveillance System (121 Cities), maintained at the Centers for Disease Control and Prevention, as an alternative data source. We fit a cyclical regression model to time series of weekly 121 Cities pneumonia and influenza deaths for 1972-1996 to estimate the excess pneumonia and influenza mortality and to compare these figures with national vital statistics estimates for 20 influenza seasons during 1972-1992. Seasonal excess mortality based on 121 Cities correlated well with the national data: for 18 (90%) of 20 seasons, our influenza epidemic severity index category approximated the result based on national vital statistics. We generated preliminary severity categories for the four recent seasons during 1992-1996. We conclude that the 121 Cities Surveillance System can be used for the timely assessment of the severity of future influenza epidemics and pandemics. Timely pneumonia and influenza mortality reporting systems established in sentinel countries worldwide would help alert public health officials and allow prompt prevention and intervention strategies during future influenza epidemics and pandemics.


Effects of endothelin on neuroeffector junction in mesenteric arteries of hypertensive rats.
The effect of endothelin, a novel vasoconstrictor peptide, on the adrenergic neuroeffector junction was investigated in isolated perfused mesenteric arteries of spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. The vasoconstrictor responses to periarterial sympathetic nerve stimulation and exogenous norepinephrine were determined. Infusion of endothelin-1 increased the baseline perfusion pressure dose dependently to similar extents in the two strains. A subpressor dose of endothelin-1 (10(-10) M) enhanced the pressor response to norepinephrine; its effect was greater in WKY rats than in SHR. Endothelin-1 (10(-12) to 10(-10) M) attenuated the pressor response to sympathetic nerve stimulation, and the degree of inhibition tended to be less in SHR than in WKY rats. Higher doses (3 x 10(-10) and 10(-9) M) of endothelin-1 enhanced the pressor response to nerve stimulation in both WKY rats and SHR. Endothelin-1 inhibited norepinephrine release from rat mesenteric arteries; the inhibition was significantly less in SHR than in WKY rats. These results suggest that endothelin enhances the responsiveness of alpha-adrenergic receptors to catecholamines, whereas it inhibits presynaptic adrenergic neurotransmission. Thus, endothelin can interact with the neuroeffector junction in addition to having a vasoconstricting effect in peripheral vessels. The difference in the mode of modulation by endothelin at the vascular neuroeffector junction in SHR from that in WKY rats might explain the maintenance of hypertension.


Invasion of the Nervous System.
In vertebrates, the nervous system (NS) is composed of a peripheral collection of neurons (the peripheral nervous system, PNS), a central set found in the brain and spinal cord (the central nervous system, CNS). The NS is protected by rather complicated multi-layer barriers that allow access to nutrients and facilitate contact with the peripheral tissues, but block entry of pathogens and toxins. Virus infections usually begin in peripheral tissues and if these barriers are weakened, they can spread into the PNS and more rarely into the CNS. Most viral infections of the NS are opportunistic or accidental pathogens that gain access via the bloodstream (e.g., HIV and various arboviruses). But a few have evolved to enter the NS efficiently by invading neurons directly and by exploiting neuronal cell biology (e.g., rhabdoviruses and alphaherpesviruses). Most NS infections are devastating and difficult to manage. Remarkably, the alphaherpesviruses establish life-long quiescent infections in the PNS, with rare but often serious CNS pathology. In this review, we will focus on how alphaherpesviruses gain access to and spread in the NS, with particular emphasis on bidirectional transport and spread within and between neurons and neural circuits, which is regulated by complex viral-host protein interactions. Finally, we will describe the wide use of alphaherpesviruses as tools to study nerve connectivity and function in animal models.


Bluetongue, epizootic haemorrhagic disease of deer and related viruses: current situation in Australia.
Since 1975 3 serotypes of bluetongue (BT) virus (BTV) have been identified in Australia: BTV1 (CSIRO156), BTV20 (CSIRO19) and BTV21. At present 2 further BT viruses (DPP90 and DPP192) have been isolated from the blood of healthy cattle in the Northern Territory (NT) and are undergoing identification. There is serological evidence for BTV15 infection in Western Australia (WA) and the NT, and a background level of serological activity to BTV serotypes 1 to 17. In addition, over 50 isolations of epizootic hemorrhagic disease of deer (EHD) viruses (EHDV) have been made and can be divided into 5 main serotypes, CSIRO157, CSIRO439, CSIRO753, CSIRO775 and DPP59. All 5 serotypes occur within the ecological range of Culicoides brevitarsis, however, as for BTV, the high prevalence of antibodies in cattle in the Kimberley region of WA and the top end of the NT suggests other vectors may also be responsible. CSIRO439 has been shown to be closely related serologically to Ibaraki virus while the other EHD viruses were distinct from Ibaraki and from each other. The complex serological relationships between BTV, EHDV and Palyam virus groups are currently being examined using serological and biochemical techniques. It is often difficult to determine the differences between BTV and EHDV serogroup members. Antibodies to both BTV and EHDV have been found in cattle, buffaloes and deer but there has been no evidence of clinical disease in any of these species or in sheep under field conditions in Australia.


[The presence of cooperative structures in tryptic fragments of troponin C].
Scanning microcalorimetry has been used to study the ability of TR-1 and TR-2 tryptic fragments of troponin C to form an ordered compact structure in solution under different conditions. It has been shown that: (1) in the presence, as well as in the absence of bivalent ions both fragments have a structure which can melt with an intensive heat absorption at heating; (2) the structure of fragment TR-1 containing two Ca2+-specific domains (domains I and II) melts as a whole under all conditions studied and therefore the domains form one cooperative block. Binding of Ca2+ or Mg2+ ions stabilizes the block structure, however, significant conformational rearrangements which would lead to a change of denaturational enthalpy do not occur; (3) Ca2+Mg2+-domains of fragment TR-2 (domains III and IV) represent individual cooperative units, blocks. Stability of these cooperative blocks strongly depends on concentration of bivalent ions and in the presence of 2 mM EDTA the melting temperature of one of them is below 10 degrees. Thermodynamic melting temperature of one of them is below 10 degrees. Thermodynamic melting parameters of cooperative blocks within peptides and in the intact molecule of troponic C are compared.


Paraoxonase activity as a tool for clinical monitoring of dogs treated for canine leishmaniasis.
This study was designed to determine if the activity of paraoxonase (PON1), an antioxidant enzyme that works as a negative acute phase reactant, is a better predictor for the clinical recovery of leishmaniotic dogs receiving standard treatments compared with inflammatory markers such as C reactive protein (CRP) and electrophoretic fractions. For this purpose we tested 20 healthy dogs (controls) and 39 leishmaniotic dogs classified as sick (group A, n=23) or severely sick (group B, n=16) and tested at admission and after 3, 7, 14, 21, 28, 35 and 42 days. At admission, CRP and electrophoresis were altered in both groups, while PON1 activity was abnormal only in group B. There were no differences related to the outcome (mortality, complications or time of recovery). PON1 activity normalized in about 2 weeks in dogs that had abnormal values at admission and a final positive outcome; CRP normalized in 4-6 weeks and electrophoretic fractions were still altered after 6 weeks. The results show that, at admission, inflammatory markers did not predict the outcome of leishmaniasis. PON1 activity decreased only in some dogs with systemic inflammation but not in those with mild leishmaniasis: when decreased, PON1 normalized earlier than other markers in dogs that responded to treatment. This finding most likely depends on the rapid decrease in oxidative phenomena. PON1 activity should therefore be tested on admission: if low values are recorded, severe inflammation may be suspected and PON1 measurement may be repeated during treatment to early identify responsive dogs.


Effectiveness of interventions for adults with psychological or emotional impairment after stroke: an evidence-based review.
This evidence-based review was conducted to evaluate the effectiveness of occupational therapy interventions to prevent or mitigate the effects of psychological or emotional impairments after stroke. Thirty-nine journal articles met the inclusion criteria. Six types of interventions were identified that addressed depression, anxiety, or mental health-related quality of life: exercise or movement based, behavioral therapy and stroke education, behavioral therapy only, stroke education only, care support and coordination, and community-based interventions that included occupational therapy. Evidence from well-conducted research supports using problem-solving or motivational interviewing behavioral techniques to address depression. The evidence is inconclusive for using multicomponent exercise programs to combat depression after stroke and for the use of stroke education and care support and coordination interventions to address poststroke anxiety. One study provided support for an intensive multidisciplinary home program in improving depression, anxiety, and health-related quality of life. The implications of the findings for practice, research, and education are discussed.


[MRI in shoulder joint instability].
The paper provides the results of MRI studies in 100 patients having complaints of pain and impaired movements in the shoulder joint in order to establish a diagnosis. Sixty-three patients were found to have MRI signs of shoulder joint instability (SJI). The paper presents and states the found MRI symptoms of SJI. The author concludes that MRI of the shoulder joint in its instability should be used appropriately as it may early reveal changes in the articular osseous, cartilaginous, and soft tissues, which is useful in diagnosing and choosing a treatment.


Use of Interventional Radiology in Critically Injured Children Admitted in a Pediatric Intensive Care Unit of a Developing Country.
Objective The aim of this study was to describe the outcome of the use of interventional radiological procedures (IRP) (angioembolization) in critically injured children. Methods A retrospective review of medical records of all children who underwent an IRP from January 2010 to December 2015 was done. Data were collected on a structured proforma and results are presented as mean with standard deviation and frequency with percentages. Result Eighteen patients were identified who underwent IRP during the study period. The mean age was 10.4 ± 4.3 years and 10 (55%) were males. Ten patients had a road traffic accident, four had a history of fall, one patient had glass cut pelvic injury, and two patients had blunt abdominal trauma, while one patient had bleeding secondary to hemipelvectomy. The genitourinary system was involved in five patients, liver in four, and spleen in two and pancreas in one patient. Bleeding was from branches of internal iliac artery in seven patients, hepatic artery in three patients, splenic artery in two patients, and middle colic artery in one patient, while one patient had blood oozing from the bone after hemi-pelvictomy. Four French vascular access sheath was placed under ultrasound guidance; this was followed by the placement of C1 catheter (Cordis, Miami, FL). After vessel identification, a 2.7F Progreat microcatheter (Terumo, Tokyo) was used for super-selective cannulation of the bleeding vessel. Intravascular coil, polyvinyl alcohol (PVA) particles, or gel foam was used for the embolization of bleeding vessels. No procedural complications were observed except minor oozing in one patient. One patient expired due to multiorgan dysfunction. Conclusion Angioembolization is a useful and relatively safe procedure in the management of vitally stable children with hemorrhagic abdominopelvic injuries. However, further studies may be needed to evaluate the efficacy and cost-effectiveness of this practice, especially in resource-constrained settings.


Regulatory space and the contextual mediation of common functional pressures: analyzing the factors that led to the German Efficiency Frontier approach.
There are no automatic links between the functional advantages and pressures associated with delegation to independent agencies for Health Technology Assessment (HTA) and their emergence in national regulatory spaces. We argue that the rise of these organizations is mediated by contextual factors, which must be explained. Accordingly, we analyze the German 'regulatory space' for health policy decision-making, identifying contextual factors relevant to the adoption of the Efficiency Frontier approach. Based on qualitative interviews with key stakeholders, we argue that the development of the Efficiency Frontier can be associated with cultural reluctance to frame healthcare prioritization decisions around cost based valuations of human health and related doubts about the validity of metrics for human health gain. Based on this finding, we conclude that the delegation of authority to independent HTA agencies follows a broadly evolutionary pattern, in which contextual factors allow for significant variation in institutional and methodological responses to the functional pressures and advantages leading to their establishment.


New oral agents for type II diabetes.
Preview Status quo obviously is not optimal in the management of patients with non-insulin-dependent diabetes, given the fact that the average hemoglobin A1c value in this diabetic population is between 9% and 10%. There is an impetus, therefore, to move up the treatment ladder more quickly than at present. The availability of new oral hypoglycemic agents provides the potential for expanding the role of this category of agents and for improving overall glycemic control in these patients.


Toward the Cessation of Seclusion and Mechanical Restraint Use in Psychiatric Hospitals: A Call for Regulatory Action.
The use of seclusion and mechanical restraints (S-R) in psychiatric hospitals remains widespread despite the traumatizing effects and risk for lethality. Neither the Centers for Medicare and Medicaid Services (CMS) nor The Joint Commission (TJC) have updated their guidelines on the use of S-R since 2005. Their regulations do not include current best practices, such as the evidence-based six core strategies (6CS) or other trauma-informed approaches, despite robust data on their effectiveness in preventing violence and S-R use. The authors describe Pennsylvania State hospitals' nearly 10-year cessation of S-R use via their continuous adherence to 6CS. In contrast, the authors describe the significant decrease in S-R use during the implementation of 6CS at a public psychiatric hospital while under U.S. Department of Justice (DOJ) monitoring and the resumption of high S-R use after DOJ monitoring and adherence to 6CS ended. The authors emphasize the importance of external regulatory oversight and mandates to safely achieve and sustain the cessation of S-R use in psychiatric hospitals. Urging CMS and TJC to update their regulations, the authors offer a roadmap to more effectively mandate the reduction and eventual cessation of S-R use in psychiatric hospitals.


[Initial kinetics of milk protein hydrolysis by chymotrypsin].
The kinetics of the initial stages of hydrolysis of alpha- and beta-caseins, alpha-lactalbumin, beta-lactoglobulin, and bovine serum albumin was studied quantitatively by electrophoresis. The hydrolysis rates of caseins (demasked polypeptide chains) exceeded the rates of the degradation of milk whey proteins limited by the protein globule demasking by more than two orders of magnitude. Even relatively accessible for hydrolysis globular proteins (beta-lactoglobulin and BSA) were hydrolyzed by the one-by-one mechanism. The maximum value of the ratio between the rates of the peptide chain demasking and the protein globule hydrolysis was 0.0035.


Association between lack of health insurance and risk of death and ESRD: results from the Kidney Early Evaluation Program (KEEP).
Uninsured adults in the United States have poor access to health care services and worse health outcomes than insured adults. Little is known about the association between lack of insurance and chronic kidney disease (CKD) progression to end-stage renal disease (ESRD) or death in patients at high risk of kidney disease. We used 2000-2011 data from the National Kidney Foundation's Kidney Early Evaluation Program (KEEP) to examine this association. The study population included KEEP participants younger than 65 years. Outcomes were time to ESRD (chronic kidney failure treated by renal replacement therapy) and time to death. Incident ESRD was ascertained by linkage to the US Renal Data System, and vital status, by linkage to the Social Security Administration Death Master File. We used Cox proportional hazard regression to examine the association between insurance and risk of death or ESRD after adjusting for demographic variables. Of 86,588 participants, 27.8% had no form of insurance, 10.3% had public insurance, and 61.9% had private insurance; 15.0% had CKD (defined as estimated glomerular filtration rate <60 mL/min/1.73 m(2) or urine albumin-creatinine ratio ≥ 30 mg/g), 63.3% had hypertension, and 27.7% had diabetes. Of participants with CKD, 29.3% had no health insurance. Participants without insurance were younger, more likely to be Hispanic and to have 12 or fewer years of education, and less likely to have seen a physician in the past year. After adjustment for demographic characteristics, uninsured KEEP participants were 82% more likely than privately insured participants to die (HR, 1.82; 95% CI, 1.56-2.12; P < 0.001) and 72% more likely to develop ESRD (HR, 1.72; 95% CI, 1.33-2.22; P < 0.001). The association between insurance and outcomes varied by CKD stage. Lack of insurance is an independent risk factor for early death and ESRD in this population at high risk of kidney disease. Considering the high morbidity and mortality and increasing cost associated with ESRD, access to appropriate health insurance coverage is warranted.


Patient satisfaction questionnaires for primary care out-of-hours services: a systematic review.
Patient satisfaction questionnaires are increasingly used for assessing quality of care. To review the evidence for the reliability and validity of patient satisfaction questionnaires for out-of-hours care. Systematic review. Primary care out-of-hours services. Searches of CINAHL, EMBASE, MEDLINE((R)) and PsycINFO using terms relevant to the measurement of patient satisfaction and out-of-hours services. Abstracts were reviewed and information relating to questionnaire content, data quality, reliability, and validity were extracted from articles by two independent researchers. Four questionnaires were found, two from the UK - the Patient Satisfaction with Out-of-Hours Care (PSOC) and Short Questionnaire for Out-of-Hours Care (SQOC) - and two from the Netherlands - the van Uden and Moll van Charante questionnaires. Questionnaire content was based on literature reviews and expert opinion; the PSOC and Moll van Charante questionnaires were also developed following interviews or focus groups with patients. Cronbach's alpha values were below 0.7 for some scales within the PSOC and van Uden questionnaires. Test-retest reliability was reported for the PSOC and Moll van Charante questionnaires. Tests of validity were few and did not give explicit consideration to the size of expected associations. Potential users wishing to assess patient satisfaction should carefully consider the content of the questionnaires and its relevance to the application and patient group. The four questionnaires have limitations relating to their development and evaluation. The PSOC and van Uden questionnaires have low levels of reliability for some scales, which should be used with caution in future surveys.


Telomerase RNA TERC and the PI3K-AKT pathway form a positive feedback loop to regulate cell proliferation independent of telomerase activity.
The core catalytic unit of telomerase comprises telomerase reverse transcriptase (TERT) and telomerase RNA (TERC). Unlike TERT, which is predominantly expressed in cancer and stem cells, TERC is ubiquitously expressed in normal somatic cells without telomerase activity. However, the functions of TERC in these telomerase-negative cells remain elusive. Here, we reported positive feedback regulation between TERC and the PI3K-AKT pathway that controlled cell proliferation independent of telomerase activity in human fibroblasts. Mechanistically, we revealed that TERC activated the transcription of target genes from the PI3K-AKT pathway, such as PDPK1, by targeting their promoters. Overexpression of PDPK1 partially rescued the deficiency of AKT activation caused by TERC depletion. Furthermore, we found that FOXO1, a transcription factor negatively regulated by the PI3K-AKT pathway, bound to TERC promoter and suppressed its expression. Intriguingly, TERC-induced activation of the PI3K-AKT pathway also played a critical role in the proliferation of activated CD4+ T cells. Collectively, our findings identify a novel function of TERC that regulates the PI3K-AKT pathway via positive feedback to elevate cell proliferation independent of telomerase activity and provide a potential strategy to promote CD4+ T cells expansion that is responsible for enhancing adaptive immune reactions to defend against pathogens and tumor cells.


General Practitioners Views of Implementing a Stratified Treatment Approach for Low Back Pain in Germany: A Qualitative Study.
The STarT Back stratified primary care approach has demonstrated clinical and cost effectiveness in the UK, and is commonly used by General Practitioners (GPs). However, it remains unknown how this approach could be implemented into the German healthcare system. The aim of this study was therefore to explore the views and perceptions of German GPs in respect to using a stratified primary care for low back pain (LBP). A 90-minute think-tank workshop was conducted with 14 male and five female GPs, during which the STarT-Back-Screening-Tool (SBST) and related research evidence was presented. This was followed by two focus groups, based on a semi-structured interview guideline to identify potential implementation barriers and opportunities. Discussions were audiotaped, transcribed and coded using a content analysis approach. For the three deductively developed main themes, 15 subthemes emerged: (1) application of the SBST, with the following subthemes: which health profession should administer it, patients known to the GP practice, the reason for the GP consultation, scoring the tool, the tool format, and the anticipated impact on GP practice; (2) psychologically informed physiotherapy, with subthemes including: provision by a physiotherapist, anticipated impact, the skills of physiotherapists, management of patients with severe psychosocial problems, referral and remuneration; (3) the management of low-risk patients, with subthemes including: concern about the appropriate advising health professional, information and media, length of consultation, and local exercise venues. The attitudes of GPs towards stratified primary care for LBP indicated positive support for pilot-testing in Germany. However, there were mixed reactions to the ability of German physiotherapists to manage high-risk patients and handle their complex clinical needs. GPs also mentioned practical difficulties in providing extended advice to low-risk patients, which nevertheless could be addressed by involvement of specifically trained medical assistants.


Suprapubic catheterization complicated by an iatrogenic enterocutaneous fistula: a case report.
Suprapubic catheterization is a common procedure employed by urologists to manage long standing voiding dysfunction or neuropathic bladders in patients. Bowel injury is a rare but important complication, in light of the consequences to patient morbidity. An 81 year old Caucasian female presented with a blocked suprapubic catheter 3 weeks after passage under cystoscopic guidance. The Foley catheter was replaced and attempted aspiration brought up a faeculent fluid. There were no signs to suggest peritonitis. Catheter was kept in situ until computed tomography imaging was obtained; this illustrated the catheter in small bowel with balloon inflated, causing partial small bowel obstruction. Patient underwent exploratory laparotomy with bowel resection, with an uneventful post-operative recovery. We speculate that the injury occurred at the time of first catheter exchange, with the tip directly piercing the small bowel. To our knowledge this particular mechanism of injury has not previously been reported. This case demonstrates the importance of remaining vigilant to iatrogenic bowel injury after cystostomy, and aids initial management if injury is suspected.


5-methylcytidylic acid: absence from mitochondrial DNA of frogs and HeLa cells.
The content of 5-methylcytidylic acid in nuclear DNA and mito chondrial DNA of Xenopus laevis and HeLa cells has been determined. Both nuclear DNA's contain 5-methylcytidylic acid. The 5-methylcytidylic acid content of X. laevis DNA is 1.7 mole percent of total nucleotides, and that of HeLc cell DNA is 0.7 mole percent. In neither mitochondrial DNA could any 5-methylcytidylic acid be detected; the limit of sensitivity was judged at below 0.1 mole percent for X. laevis DNA and below 0.05 mole percent for HeLa cell DNA.


The effect of endometrial injury on ongoing pregnancy rate in unselected subfertile women undergoing in vitro fertilization: a randomized controlled trial.
Does endometrial injury in the cycle preceding ovarian stimulation for in vitro fertilization (IVF) improve the ongoing pregnancy rate in unselected subfertile women? Endometrial injury induced by endometrial aspiration in the preceding cycle does not improve the ongoing pregnancy rate in unselected subfertile women undergoing IVF. Implantation failure remains one of the major limiting factors for IVF success. Mechanical endometrial injury in the cycle preceding ovarian stimulation of IVF treatment has been shown to improve implantation and pregnancy rates in women with repeated implantation failures. There is limited data on unselected subfertile women, especially those undergoing their first IVF treatment. This randomized controlled trial recruited 300 unselected subfertile women scheduled for IVF/ICSI treatment between March 2011 and August 2013. Subjects were randomized into endometrial aspiration (EA) (n = 150) and non-EA (n = 150) groups according to a computer-generated randomization list. Subjects were recruited and randomized in the assisted reproductive unit at the University of Hong Kong. In the preceding cycle, women in the EA group underwent endometrial aspiration using a Pipelle catheter in mid-luteal phase. All women were treated with a cycle of IVF/ICSI. Pregnancy outcomes were compared. There were no significant differences in baseline or cycle characteristics between the groups. There were 209 subjects (69.7%) who were undergoing their first IVF cycle and 91 (30.3%) subjects who had repeated cycles. There was no significant difference in ongoing pregnancy rates [26.7% (40/150) versus 32.0% (48/150); RR 0.833 (95% CI 0.585-1.187), P = 0.375] in the EA and non-EA groups. The implantation rates [32.8% (67/204) versus 29.7% (68/229); RR 1.080 (95% CI 0.804-1.450), P = 0.120], clinical pregnancy rates [34.0% (51/150) versus 38.0 (57/150); RR 0.895 (95% CI 0.661-1.211), P = 0.548], miscarriage rates [30.3% (17/56) versus 18.6% (11/59), RR 1.628 (95% CI 0.838-3.164), P = 0.150] and multiple pregnancy rates [31.3% (16/51) versus 19.3% (11/57), RR 1.626 (95% CI 0.833-3.172), P = 0.154] were all comparable between the EA and non-EA groups. Subgroup analysis in women having first embryo transfer (n = 209) also demonstrated no significant difference in ongoing pregnancy rates, but for women undergoing repeated cycles (n = 91), the on-going pregnancy rate was significantly lower in the EA group than in the non-EA group. The study aimed at assessing an unselected population of subfertile women by recruiting consecutive women attending our fertility clinic. However, since the majority of the recruited women (69.7%) were having their first IVF treatments, the results may not be generalizable to all women undergoing IVF. Previous RCTs and meta-analyses have suggested improved pregnancy rates after pretreatment endometrial injury in women with repeated implantation failure. A recent RCT also showed increased pregnancy rates in unselected subfertile women after endometrial injury, although that study was terminated early and thus underpowered. Our study showed with adequate power that no significant improvement in pregnancy rates was observed after endometrial injury in unselected women undergoing IVF treatment. The study was supported by the Small Project Funding 201309176012 of the Committee on Research and Conference Grants, University of Hong Kong. The authors have nothing to disclose. HKCTR-1646 and NCT 01977976.


Severity-associated markers and assessment model for predicting the severity of COVID-19: a retrospective study in Hangzhou, China.
The severity of COVID-19 associates with the clinical decision making and the prognosis of COVID-19 patients, therefore, early identification of patients who are likely to develop severe or critical COVID-19 is critical in clinical practice. The aim of this study was to screen severity-associated markers and construct an assessment model for predicting the severity of COVID-19. 172 confirmed COVID-19 patients were enrolled from two designated hospitals in Hangzhou, China. Ordinal logistic regression was used to screen severity-associated markers. Least Absolute Shrinkage and Selection Operator (LASSO) regression was performed for further feature selection. Assessment models were constructed using logistic regression, ridge regression, support vector machine and random forest. The area under the receiver operator characteristic curve (AUROC) was used to evaluate the performance of different models. Internal validation was performed by using bootstrap with 500 re-sampling in the training set, and external validation was performed in the validation set for the four models, respectively. Age, comorbidity, fever, and 18 laboratory markers were associated with the severity of COVID-19 (all P values < 0.05). By LASSO regression, eight markers were included for the assessment model construction. The ridge regression model had the best performance with AUROCs of 0.930 (95% CI, 0.914-0.943) and 0.827 (95% CI, 0.716-0.921) in the internal and external validations, respectively. A risk score, established based on the ridge regression model, had good discrimination in all patients with an AUROC of 0.897 (95% CI 0.845-0.940), and a well-fitted calibration curve. Using the optimal cutoff value of 71, the sensitivity and specificity were 87.1% and 78.1%, respectively. A web-based assessment system was developed based on the risk score. Eight clinical markers of lactate dehydrogenase, C-reactive protein, albumin, comorbidity, electrolyte disturbance, coagulation function, eosinophil and lymphocyte counts were associated with the severity of COVID-19. An assessment model constructed with these eight markers would help the clinician to evaluate the likelihood of developing severity of COVID-19 at admission and early take measures on clinical treatment.


Health seeking behaviour and health service utilization in Pakistan: challenging the policy makers.
There is a growing literature on health seeking behaviours and the determinants of health services utilization especially in the context of developing countries. However, very few focused studies have been seen in Pakistan in this regard. This paper presents an extensive literature review of the situation in developing countries and relates the similar factors responsible for shaping up of a health seeking behaviour and health service utilization in Pakistan. The factors determining the health behaviours may be seen in various contexts: physical, socio-economic, cultural and political. Therefore, the utilization of a health care system, public or private, formal or non-formal, may depend on socio-demographic factors, social structures, level of education, cultural beliefs and practices, gender discrimination, status of women, economic and political systems environmental conditions, and the disease pattern and health care system itself. Policy makers need to understand the drivers of health seeking behaviour of the population in an increasingly pluralistic health care system. Also a more concerted effort is required for designing behavioural health promotion campaigns through inter-sectoral collaboration focusing more on disadvantaged segments of the population.


Unmanipulated haploidentical blood and marrow transplantation: where we are.
Human leukocyte antigen (HLA)-mismatched/haploidentical blood and marrow transplants (haplo-BMT) from family donors have been intensively studied because of the decreasing family size in mainland China, and also because the Chinese Marrow Donor Program is still not big enough. The protocol for unmanipulated haplo-BMT has been designated as 'GIAC' by Dr DP Lu--'G' represents granulocyte colony-stimulating factor mobilisation; 'I' stands for immunosuppression during pre-conditioning being prolonged and intensified; 'A' stands for the use of antithymocyte globulin; 'C' means combined use of bone marrow and peripheral blood as the graft. Haplo-BMT with GIAC regimen has been shown to be feasible for many applications as reported in 2004. Under this protocol, haplo-BMT has achieved comparable outcomes in terms of severe acute graft-versus-host disease (GVHD), chronic GVHD, relapse, treatment-related mortality (TRM), disease-free survival (DFS), and overall survival with HLA-identical sibling transplantation. The probabilities of DFS at 2 years in haplo-BMT setting were 70.7%, 49.6%, 22.2% in standard-risk, high-risk, advanced disease groups, respectively. As the third party cells, cord blood co-infusion could significantly reduce the incidence and severity of acute GVHD, and also 100-day TRM. The majority of refractory cytomegalovirus, Epstein-Barr virus and aspergillus infections can be controlled by adoptive cellular therapy. Many patients who early relapsed after BMT and failed, or are ineligible for standard therapy, have been salvaged with dendritic cell-primed cytokine-induced killer cells. With these new strategies, the lower TRM and improved DFS have been attained. Therefore, it is better to consider haplo-BMT for the patients with otherwise incurable haematological malignancies at earlier stage, when matched sibling or unrelated donors are not available.


Mutations Impairing GSK3-Mediated MAF Phosphorylation Cause Cataract, Deafness, Intellectual Disability, Seizures, and a Down Syndrome-like Facies.
Transcription factors operate in developmental processes to mediate inductive events and cell competence, and perturbation of their function or regulation can dramatically affect morphogenesis, organogenesis, and growth. We report that a narrow spectrum of amino-acid substitutions within the transactivation domain of the v-maf avian musculoaponeurotic fibrosarcoma oncogene homolog (MAF), a leucine zipper-containing transcription factor of the AP1 superfamily, profoundly affect development. Seven different de novo missense mutations involving conserved residues of the four GSK3 phosphorylation motifs were identified in eight unrelated individuals. The distinctive clinical phenotype, for which we propose the eponym Aymé-Gripp syndrome, is not limited to lens and eye defects as previously reported for MAF/Maf loss of function but includes sensorineural deafness, intellectual disability, seizures, brachycephaly, distinctive flat facial appearance, skeletal anomalies, mammary gland hypoplasia, and reduced growth. Disease-causing mutations were demonstrated to impair proper MAF phosphorylation, ubiquitination and proteasomal degradation, perturbed gene expression in primary skin fibroblasts, and induced neurodevelopmental defects in an in vivo model. Our findings nosologically and clinically delineate a previously poorly understood recognizable multisystem disorder, provide evidence for MAF governing a wider range of developmental programs than previously appreciated, and describe a novel instance of protein dosage effect severely perturbing development.


Preparation and characterization of whey protein film incorporated with TiO2 nanoparticles.
Biodegradable titanium dioxide (TiO(2))/whey protein isolate (WPI) blend films were made by casting denatured WPI film solutions incorporated with TiO(2) nanoparticles. X-ray diffraction, UV-vis spectra, and fluorescence spectra of the films showed the successful incorporation of TiO(2) nanoparticles into the WPI matrix and indicated the interactions between TiO(2) and WPI. Mechanical tests revealed the antiplasticizing effect of TiO(2) nanoparticles on the WPI/TiO(2) film. Small amounts (<1 wt%) of added TiO(2) nanoparticles significantly increase the tensile properties of WPI film, but also decrease the moisture barrier properties. The addition of higher amounts (>1 wt%) of TiO(2) improves moisture barrier properties but lowers the tensile properties of the film. Microstructural evaluation confirmed the aggregation and distribution of TiO(2) nanoparticles within the WPI matrix and validated the results of functional properties of the WPI/TiO(2) film.


Performance evaluation of a sensorless adaptive optics multiphoton microscope.
A wavefront sensorless adaptive optics technique was combined with a custom-made multiphoton microscope to correct for specimen-induced aberrations. A liquid-crystal-on-silicon (LCoS) modulator was used to systematically generate Zernike modes during image recording. The performance of the instrument was evaluated in samples providing different nonlinear signals and the benefit of correcting higher order aberrations was always noticeable (in both contrast and resolution). The optimum aberration pattern was stable in time for the samples here involved. For a particular depth location within the sample, the wavefront to be precompensated was independent on the size of the imaged area (up to ∼ 360 × 360 μm(2)). The mode combination optimizing the recorded image depended on the Zernike correction control sequence; however, the final images hardly differed. At deeper locations, a noticeable dominance of spherical aberration was found. The influence of other aberration terms was also compared to the effect of the spherical aberration.


A micronised, dispersible ferric pyrophosphate with high relative bioavailability in man.
Ferric pyrophosphate is a water-insoluble Fe compound used to fortify infant cereals and chocolate-drink powders as it causes no organoleptic changes to the food vehicle. However, it is only of low absorption in man. Recently, an innovative ferric pyrophosphate has been developed (Sunactive Fe trade mark ) based on small-particle-size ferric pyrophosphate (average size 0.3 microm) mixed with emulsifiers, so that it remains in suspension in liquid products. The aim of the present studies was to compare Fe absorption of micronised, dispersible ferric pyrophosphate (Sunactive Fe trade mark ) with that of ferrous sulfate in an infant cereal and a yoghurt drink. Two separate Fe absorption studies were made in adult women (ten women/study). Fe absorption was based on the erythrocyte incorporation of stable isotopes ((57)Fe and (58)Fe) 14 d after the intake of labelled test meals of infant cereal (study 1) or yoghurt drink (study 2). Each test meal was fortified with 5 mg Fe as ferrous sulfate or micronised, dispersible ferric pyrophosphate. Results are presented as geometric means. There was no statistically significant difference between Fe absorption from micronised, dispersible ferric pyrophosphate- and ferrous sulfate-fortified infant cereal (3.4 and 4.1 % respectively; P=0.24) and yoghurt drink (3.9 and 4.2 % respectively; P=0.72). The results of the present studies show that micronised, dispersible ferric pyrophosphate is as well absorbed as ferrous sulfate in adults. The high relative Fe bioavailability of micronised, dispersible ferric pyrophosphate indicates the potential usefulness of this compound for food fortification.


Localization of kala-azar in the endemic region of Bihar, India based on land use/land cover assessment at different scales.
Land cover, a critical variable in the epidemiology of kala-azar, can be remotely characterized by customizing and integrating "state-of-the-art" imagery at different spatial scales from different sensors onboard satellites. A study was conducted at four levels (national, state, district and village) investigating the role of land use/land cover (LULC) for leishmaniasis transmission resulting in a framework highlighting the links between LULC and areas endemic for the disease. Distribution maps were analysed by a probabilistic approach (Bayesian classifier) which produced a set of "suitability estimates" based on the probability of sand fly presence. The development of a sound knowledge of each link in the predicted sequence of satellite views offering an extraordinary opportunity to support the mapping of kala-azar endemicity and stratification of areas suitable for sand fly habitats across the country as well as at the local scale.


Broadly tunable wideband optical single sideband generation using self-coupled silicon resonator.
We propose and demonstrate a tunable broadband optical single sideband generation using self-coupled silicon micro-ring resonator. We exploit self-coupling in a ring cavity to generate tunable resonance splitting. Using the proposed device, single sideband with carrier signal is generated from a double sideband signal. Experimental verification of power fading free transmission through a 43 km signal mode optical fiber is achieved for an RF frequency range of 1-20 GHz, extendible to higher frequencies. We also achieved a spurious free dynamic range of > 99.9±1.05 dB.Hz2/3 over the demonstrated frequency range. Furthermore, error-free data transmission of 1-12 Gbps over a 43 km fiber is also demonstrated with a detailed analysis of bit error rate as well.


Development of fulminant Type 1 diabetes with thrombocytopenia after influenza vaccination: a case report.
Fulminant Type 1 diabetes was originally reported as idiopathic Type 1 diabetes. Involvement of viral infections in the pathogenesis of fulminant T1D has been suggested, but the development of fulminant Type 1 diabetes after influenza vaccination has not been reported. We report a case of fulminant Type 1 diabetes with thrombocytopenia following influenza vaccination. A 54-year-old man was admitted to hospital with hyperglycaemia and diabetic ketosis. Seven days before admission, he received a seasonal influenza vaccine for the prevention of influenza infection. On admission, blood glucose was 29 mmol/L and HbA1c 40 mmol/mol (5.9%). Fasting and 2-h C-peptide immunoreactivity were <0.0333 nmol/L and 0.0999 nmol/L, respectively. Anti-GAD and anti-IA-2 antibodies were negative, so no autoimmunity seemed to participate in the etiology. ELISPOT assay also showed no association with T cell-mediated autoimmunity. HLA genotypes were consistent with susceptibility to fulminant Type 1 diabetes. After the abrupt onset of diabetes, he showed mild thrombocytopenia, which has been observed for approximately 5 years after diabetes development. This is the first description of fulminant Type 1 diabetes after influenza vaccination. Our observation raises the possibility that influenza vaccination might trigger this condition via the TLR7 pathway.


Differences in Patient Experience Between Hispanic and Non-Hispanic White Patients Across U.S. Hospitals.
Despite the increased emphasis on patient experience, little is known about whether there are meaningful differences in hospital satisfaction between Hispanic and non-Hispanic whites. To determine if satisfaction differs, we used Hospital Consumer Assessment of Healthcare Providers and Systems (HCAHPS) survey data (2009-2010) reported by hospitals to compare responses between Hispanic and non-Hispanic white patients. Clustered logistic regression models identified within-hospital and between-hospital differences in satisfaction. Of the 3,864,938 respondents, 6.2% were Hispanics, who were more often younger and females and less likely to have graduated from high school. Hispanics were overall more likely to recommend their hospital (74.1% vs. 70.9%, p < .001) and to rate it 9 or 10 (72.5% vs. 65.9%, p < .001) than whites. Increased satisfaction among Hispanics was more pronounced when compared with whites within the same hospitals, with significantly higher ratings on all HCAHPS measures. However, hospitals serving a higher percentage of Hispanics had lower satisfaction scores for both Hispanic and white patients than other hospitals. There were significant but only modest-sized differences in patient experience between Hispanic and white patients across U.S. hospitals. Hispanics tended to be more satisfied with their care but received care at lower-performing hospitals.


Enterococcal bacteraemia: factors influencing mortality, length of stay and costs of hospitalization.
Enterococci are a major cause of nosocomial bacteraemia. The impacts of vanB vancomycin resistance and antibiotic therapy on outcomes in enterococcal bacteraemia are unclear. Factors that affect length of stay (LOS) and costs of managing patients with enterococcal bacteraemia are also unknown. This study aimed to identify factors associated with mortality, LOS and hospitalization costs in patients with enterococcal bacteraemia and the impact of vancomycin resistance and antibiotic therapy on these outcomes. Data from 116 patients with vancomycin-resistant Enterococci (VRE), matched 1:1 with patients with vancomycin-susceptible Enterococcus (VSE), from two Australian hospitals were reviewed for clinical and economic outcomes. Univariable and multivariable logistic and quantile regression analyses identified factors associated with mortality, LOS and costs. Intensive care unit admission (OR, 8.57; 95% CI, 3.99-18.38), a higher burden of co-morbidities (OR, 4.55; 95% CI, 1.83-11.33) and longer time to appropriate antibiotics (OR, 1.02; 95% CI, 1.01-1.03) were significantly associated with mortality in enterococcal bacteraemia. VanB vancomycin resistance increased LOS (4.89 days; 95% CI, 0.56-11.52) and hospitalization costs (AU$ 28 872; 95% CI, 734-70 667), after adjustment for confounders. Notably, linezolid definitive therapy was associated with lower mortality (OR, 0.13; 95% CI, 0.03-0.58) in vanB VRE bacteraemia patients. In patients with VSE bacteraemia, time to appropriate antibiotics independently influenced mortality, LOS and hospitalization costs, and underlying co-morbidities were associated with mortality. The study findings highlight the importance of preventing VRE bacteraemia and the significance of time to appropriate antibiotics in the management of enterococcal bacteraemia.


Thirteen Years of Phleboviruses Circulation in Lombardy, a Northern Italy Region.
Phleboviruses transmitted by phlebotomine sandflies are endemic in the Mediterranean basin. Toscana phlebovirus (TOSV), Sicilian phlebovirus (SFSV), and Naples phlebovirus (SFNV) are responsible of summer fever, with well-known pathogenic potential for humans ranging from asymptomatic to mild fever, in addition to neuro-invasive infections during summer. Although TOSV, in particular, is a significant and well-known human pathogen, SFVs remain neglected, with many gaps in the relevant knowledge. Sero-epidemiological studies and case reports recently showed a geographical wider distribution than previously considered, although the real incidence of phleboviruses infections in the Mediterranean area is still unknown. Here we retrospectively evaluated the circulation of phleboviruses during summer seasons between 2007 and 2019 in 649 patients showing neurological symptoms using both molecular and serological approaches. We found that 42/649 (6.5%) subjects experienced phlebovirus infection and only 10/42 cases were detected by molecular assays, whereas the other 32/42 were identified using serological approaches, including neutralization assays. During the 2013 summer, an outbreak in the Lombardy region is described because the prevalence of phlebovirus infection reached 37.2% (19/51 subjects). Interestingly, only 5/19 (26.5%) reported traveling in endemic areas. Of note, no cross-neutralization was observed between different strains tested, showing the possibility to be reinfected by newly discovered phlebovirus strains. In conclusion, phlebovirus infections are still inadequately considered by physicians and are generally underestimated. However, based on our results, sandfly fever viruses should be routinely included in diagnostic panels during summer period, including in Northern Italy.


Evaluation of mesotherapeutic injections of three different combinations of lipolytic agents for body contouring.
There has been significant interest recently in the technique of mesotherapy as a method of 'melting fat' for body contouring. The aim of this study was to evaluate the safety, efficacy and lipolytic potential of several compounds commonly used in cosmetic mesotherapy. A total of 75 women (mean age: 33 years) were separated randomly into three mesotherapy groups. Injections were performed for 15 treatments once a week. The main component of each cocktail was phosphatidylcholine/deoxycholic acid for group 1, caffeine for group 2, and Conjonctyl® for group 3. Outcome was evaluated by weight, body fat percentage (BFP), circumference measurements, and patient questionnaires. Seventy-two of all the patients (96%) showed a circumference loss. An average circumference reduction of 4.41 cm per site for group 1, 2.99 cm for group 2, and 2.10 cm for group 3 was achieved. Mean body circumference loss was statistically significant, with p < 0.00. Weight loss was 5.33 ± 1.09 kg for group 1, 3.74 ± 1.51 kg for group 2, and 2.82 ± 1.43 kg for group 3. Seventy-four subjects (98.7%) showed a BFP decrease. A questionnaire indicated high patient satisfaction (63%). No patient showed irregularities, dimples or any serious side effects after treatment. Mesotherapy is a well-tolerated and effective alternative treatment modality for reducing the diameter of body circumference.


Multicenter Outbreak of Gram-Negative Bloodstream Infections in Hemodialysis Patients.
Contaminated water and other fluids are increasingly recognized to be associated with health care-associated infections. We investigated an outbreak of Gram-negative bloodstream infections at 3 outpatient hemodialysis facilities. Matched case-control investigations. Patients who received hemodialysis at Facility A, B, or C from July 2015 to November 2016. Infection control practices, sources of water, dialyzer reuse, injection medication handling, dialysis circuit priming, water and dialysate test findings, environmental reservoirs such as wall boxes, vascular access care practices, pulsed-field gel electrophoresis, and whole-genome sequencing of bacterial isolates. Cases were defined by a positive blood culture for any Gram-negative bacteria drawn July 1, 2015 to November 30, 2016 from a patient who had received hemodialysis at Facility A, B, or C. Exposures in cases and controls were compared using matched univariate conditional logistic regression. 58 cases of Gram-negative bloodstream infection occurred; 48 (83%) required hospitalization. The predominant organisms were Serratia marcescens (n=21) and Pseudomonas aeruginosa (n=12). Compared with controls, cases had higher odds of using a central venous catheter for dialysis (matched odds ratio, 54.32; lower bound of the 95% CI, 12.19). Facility staff reported pooling and regurgitation of waste fluid at recessed wall boxes that house connections for dialysate components and the effluent drain within dialysis treatment stations. Environmental samples yielded S marcescens and P aeruginosa from wall boxes. S marcescens isolated from wall boxes and case-patients from the same facilities were closely related by pulsed-field gel electrophoresis and whole-genome sequencing. We identified opportunities for health care workers' hands to contaminate central venous catheters with contaminated fluid from the wall boxes. Limited patient isolates for testing, on-site investigation occurred after peak of infections. This large outbreak was linked to wall boxes, a previously undescribed source of contaminated fluid and biofilms in the immediate patient care environment.


QSAR study for the soybean 15-lipoxygenase inhibitory activity of organosulfur compounds derived from the essential oil of garlic.
In this study, multiple linear regression (MLR) and partial least-squares (PLS) techniques were used for modeling the soybean 15-lipoxygenase inhibitory activity of a varied group of mono-, di-, and trisulfides derived from the essential oil of garlic. The structures of the compounds under study were characterized by means of calculated physicochemical parameters and several nonempirical descriptors, such as topological, geometrical, and quantum chemical indices. The results obtained indicate that the inhibitory activity is strongly dependent on the ability of the compounds to participate in dispersive interactions with the enzyme, as expressed by the solvent-accessible surface area (SASA) and the average distance/distance degree descriptor (ADDD) index. On the other hand, the high contribution of the lowest unoccupied molecular orbit term (LUMO) in the PLS models derived for the di- and trisulfides suggests that the solute's electron-acceptor capacity plays a fundamental role in the inhibitory activity exhibited for these compounds. Finally, the geometric features as expressed by the shape parameters included in the models indicate a low but not negligible positive contribution of molecular linearity in the enzyme-inhibitor binding. In summary, the developed quantitative structure-activity relationship approach successfully accounts for the potencies of organosulfur compounds acting on soybean 15-lipoxygenase and thereby offers both a guide for the synthesis of new compounds and a hypothesis for the molecular basis of their activity.


Synthesis and tau RNA binding evaluation of ametantrone-containing ligands.
We describe the synthesis and characterization of ametantrone-containing RNA ligands based on the derivatization of this intercalator with two neamine moieties (Amt-Nea,Nea) or with one azaquinolone heterocycle and one neamine (Amt-Nea,Azq) as well as its combination with guanidinoneamine (Amt-NeaG4). Biophysical studies revealed that guanidinylation of the parent ligand (Amt-Nea) had a positive effect on the binding of the resulting compound for Tau pre-mRNA target as well as on the stabilization upon complexation of some of the mutated RNA sequences associated with the development of tauopathies. Further studies by NMR revealed the existence of a preferred binding site in the stem-loop structure, in which ametantrone intercalates in the characteristic bulged region. Regarding doubly-functionalized ligands, binding affinity and stabilizing ability of Amt-Nea,Nea were similar to those of the guanidinylated ligand, but the two aminoglycoside fragments seem to interfere with its accommodation in a single binding site. However, Amt-Nea,Azq binds at the bulged region in a similar way than Amt-NeaG4. Overall, these results provide new insights on fine-tuning RNA binding properties of ametantrone by single or double derivatization with other RNA recognition motifs, which could help in the future design of new ligands with improved selectivity for disease-causing RNA molecules.


Extremely Low Resistance of Li3PO4 Electrolyte/Li(Ni0.5Mn1.5)O4 Electrode Interfaces.
Solid-state Li batteries containing Li(Ni0.5Mn1.5)O4 as a 5 V-class positive electrode are expected to revolutionize mobile devices and electric vehicles. However, practical applications of such batteries are hampered by the high resistance at their solid electrolyte/electrode interfaces. Here, we achieved an extremely low electrolyte/electrode interface resistance of 7.6 Ω cm2 in solid-state Li batteries with Li(Ni0.5Mn1.5)O4. Furthermore, we observed spontaneous migration of Li ions from the solid electrolyte to the positive electrode after the formation of the electrolyte/electrode interface. Finally, we demonstrated stable fast charging and discharging of the solid-state Li batteries at a current density of 14 mA/cm2. These results provide a solid foundation to understand and fabricate low-resistance electrolyte/electrode interfaces.


Macrophage-associated responses to chrysotile.
The different pulmonary macrophage (airway macrophages, alveolar macrophages, interstitial macrophages, intravascular macrophages, pleural macrophages) are an important part of the lungs' defences against non-fibrous and fibrous particles deposited by inhalation. The first line of defence is airway macrophages and alveolar macrophages (AM) which initially interact with deposited chrysotile fibres and subsequently release a number of mediators including growth regulatory and chemotactic proteins, arachidonic acid metabolites, proteases, NO and active oxygen species, all of which can affect--also adversely--specific target cells in the lung. Mechanical clearance via the mucociliary escalator and dissolution of phagocytized fibres in the acidic milieu of the phagolysosome in pulmonary macrophages are further important functions of AM. Chrysotile appears to be more toxic or at least has the same toxicity to AM as amphibole fibres when doses of a similar mass are administered. However, on a fibre number basis chrysotile appears to be less toxic to AM. The importance of the appropriate dose parameter--i.e. fibre mass, number or surface area--needs to be considered in in vitro as well as in in vivo studies. Short chrysotile fibres are cleared from rat lungs very rapidly whereas longer ones are cleared at a much slower rate. This is due to efficient phagocytosis of short fibres by AM accompanied by dissolution in the acidic milieu of the phagolysosome. Prediction of chrysotile clearance in primate lung based on results from rat studies result in an overall retention half-time of approximately 105 days, based on which no long-term accumulation of chrysotile in the primate lung is to be expected. Long-term inhalation studies in baboons exposed to chrysotile confirm the very fast build up of a low steady-state lung burden, consistent with a pulmonary retention half-time for chrysotile of approximately 90 days. Despite the fast clearance and low pulmonary accumulation of chrysotile, the resulting effects, such as asbestosis, were found to be of the same severity in rats as those induced by amphibole exposure. In the amphibole-exposed rats, the fibre lung burden continued to increase with exposure time. The potential contamination of chrysotile with tremolite cannot explain these results since there was no increased pulmonary accumulation of fibres in the chrysotile-exposed rats. Effects due to lung particle overload are not to be expected in long-term chrysotile inhalation studies since no accumulation occurs and one needs to distinguish clearly between a volumetric overload of the AM (not occurring with chrysotile) and failed or partial phagocytosis of long fibres.(ABSTRACT TRUNCATED AT 400 WORDS)


Msp1/ATAD1 in Protein Quality Control and Regulation of Synaptic Activities.
Mitochondrial function depends on the efficient import of proteins synthesized in the cytosol. When cells experience stress, the efficiency and faithfulness of the mitochondrial protein import machinery are compromised, leading to homeostatic imbalances and damage to the organelle. Yeast Msp1 (mitochondrial sorting of proteins 1) and mammalian ATAD1 (ATPase family AAA domain-containing 1) are orthologous AAA proteins that, fueled by ATP hydrolysis, recognize and extract mislocalized membrane proteins from the outer mitochondrial membrane. Msp1 also extracts proteins that have become stuck in the import channel. The extracted proteins are targeted for proteasome-dependent degradation or, in the case of mistargeted tail-anchored proteins, are given another chance to be routed correctly. In addition, ATAD1 is implicated in the regulation of synaptic plasticity, mediating the release of neurotransmitter receptors from postsynaptic scaffolds to allow their trafficking. Here we discuss how structural and functional specialization imparts the unique properties that allow Msp1/ATAD1 ATPases to fulfill these diverse functions and also highlight outstanding questions in the field.


Organizational capacity for patient and family engagement in hospital planning and improvement: interviews with patient/family advisors, managers and clinicians.
Patient and family engagement (PE) in healthcare planning and improvement achieves beneficial outcomes and is widely advocated, but a lack of resources is a critical barrier. Little prior research studied how organizations support engagement specifically in hospitals. We explored what constitutes hospital capacity for engagement. We conducted descriptive qualitative interviews and complied with criteria for rigour and reporting in qualitative research. We interviewed patient/family advisors, engagement managers, clinicians and executives at hospitals with high engagement activity, asking them to describe essential resources or processes. We used content analysis and constant comparison to identify themes and corresponding quotes and interpreted findings by mapping themes to two existing frameworks of PE capacity not specific to hospitals. We interviewed 40 patient/family advisors, patient engagement managers, clinicians and corporate executives from nine hospitals (two < 100 beds, four 100 + beds, three teaching). Four over-arching themes about capacity considered essential included resources, training, organizational commitment and staff support. Views were similar across participant and hospital groups. Resources included funding and people dedicated to PE and technology to enable communication and collaboration. Training encompassed initial orientation and project-specific training for patient/family advisors and orientation for new staff and training for existing staff on how to engage with patient/family advisors. Organizational commitment included endorsement from the CEO and Board, commitment from staff and continuous evaluation and improvement. Staff support included words and actions that conveyed value for the role and input of patient/family advisors. The blended, non-hospital-specific framework captured all themes. Hospitals of all types varied in the availability of funding dedicated to PE. In particular, reimbursement of expenses and compensation for time and contributions were not provided to patient/family advisors. In addition to skilled engagement managers, the role of clinician or staff champions was viewed as essential. The findings build on prior research that largely focused on PE in individual clinical care or research or in primary care planning and improvement. The findings closely aligned with existing frameworks of organizational capacity for PE not specific to hospital settings, which suggests that hospitals could use the blended framework to plan, evaluate and improve their PE programs. Further research is needed to yield greater insight into how to promote and enable compensation for patient/family advisors and the role of clinician or staff champions in supporting PE.


Crystallization of free cholesterol in model macrophage foam cells.
-The present study examined free cholesterol (FC) crystallization in macrophage foam cells. Model foam cells (J774 or mouse peritoneal macrophages [MPMs]) were incubated with acetylated low density lipoprotein and FC/phospholipid dispersions for 48 hours, resulting in the deposition of large stores of cytoplasmic cholesteryl esters (CEs). The model foam cells were then incubated for up to 5 days with an acyl-coenzyme A:cholesterol acyltransferase (ACAT) inhibitor (CP-113,818) in the absence of an extracellular FC acceptor to allow intracellular accumulation of FC. FC crystals of various shapes and sizes formed in the MPMs but not in the J774 macrophages. Examination of the MPM monolayers by microscopy indicated that the crystals were externalized rapidly after formation and thereafter continued to increase in size. Incubating J774 macrophages with 8-(4-chlorophenylthio)adenosine 3':5'-cyclic monophosphate (CPT-cAMP) in addition to CP-113,818 caused FC crystal formation as a consequence of CPT-cAMP stimulation of CE hydrolysis and inhibition of cell growth. In addition, 2 separate cholesterol phases (liquid-crystalline and cholesterol monohydrate) in the plane of the membrane bilayer were detected after 31 hours of ACAT inhibition by the use of small-angle x-ray diffraction of J774 macrophage foam cells treated with CPT-cAMP. Other compounds reported to inhibit ACAT, namely progesterone (20 microgram/mL) and N-acetyl-D-sphingosine (c(2)-ceramide, 10 microgram/mL), induced cellular toxicity in J774 macrophage foam cells and FC crystallization when coincubated with CPT-cAMP. Addition of the extracellular FC acceptors apolipoproteins (apo) E and A-I (50 microgram/mL) reduced FC crystal formation. In MPMs, lower cell density and frequent changes of medium were conducive to crystal formation. This may be due to "dilution" of apoE secreted by the MPMs and is consistent with our observation that the addition of exogenous apoE or apoA-I inhibits FC crystal formation in J774 macrophage foam cells cotreated with CP-113,818 plus CPT-cAMP. These data demonstrate that FC crystals can form from the hydrolysis of cytoplasmic stores of CEs in model foam cells. FC crystal formation can be modulated by the addition of extracellular FC acceptors or by affecting the cellular rate of CE hydrolysis. This process may contribute to the formation of FC crystals in atherosclerotic plaques.


Reliability over time of EEG-based mental workload evaluation during Air Traffic Management (ATM) tasks.
Machine-learning approaches for mental workload (MW) estimation by using the user brain activity went through a rapid expansion in the last decades. In fact, these techniques allow now to measure the MW with a high time resolution (e.g. few seconds). Despite such advancements, one of the outstanding problems of these techniques regards their ability to maintain a high reliability over time (e.g. high accuracy of classification even across consecutive days) without performing any recalibration procedure. Such characteristic will be highly desirable in real world applications, in which human operators could use such approach without undergo a daily training of the device. In this work, we reported that if a simple classifier is calibrated by using a low number of brain spectral features, between those ones strictly related to the MW (i.e. Frontal and Occipital Theta and Parietal Alpha rhythms), those features will make the classifier performance stable over time. In other words, the discrimination accuracy achieved by the classifier will not degrade significantly across different days (i.e. until one week). The methodology has been tested on twelve Air Traffic Controls (ATCOs) trainees while performing different Air Traffic Management (ATM) scenarios under three different difficulty levels.


Friends, lovers and spouses: intimacy in young adults.
Intimacy in young heterosexual adults was studied as a function of their familial roles. The 168 males and females employed represented four familial role groups: late adolescents, single adults, married people and parents. Participants were administered two forms of an Intimacy Scale (Sharabany, 1994) in which they described their desired and their obtained intimacy with a same-sex and an opposite-sex best friend. Results indicated that (a) intimacy of adults with opposite-sex partner was higher than intimacy with same-sex friend. (b) Although no direct effect of familial role on intimacy was found, the married and parent groups displayed greater intimacy towards their spouses than late adolescents and single adults towards their opposite-sex partners. (c) Women who were late adolescents and women who were married scored significantly higher than men in intimacy. However, single women expressed significantly lower intimacy than single men. (d) Higher intimacy with opposite-sex partner was associated with a concurrent lower same-sex intimacy. (e) Satisfaction with other-sex partner was higher in the married group than in the other groups.


Optimizing HIV transition services for young adults.
A total of 1.8 million adolescents live with HIV and constitute the only age group in which HIV-associated mortality continues to rise. Meeting their healthcare needs as they transition from paediatric services to adult care is vital for their own health and in prevention of onward transmission to partners and offspring. In this review, we discuss the issues around transition, both within HIV health care and the wider transitions adolescents negotiate as they move out of childhood and into adult life. Although transition models vary widely across the globe, the difficulties faced in gathering robust outcome data following transition to adult care and in linkage between paediatric and adult cohorts are universal. Data are particularly sparse for behaviourally infected adolescents outside North America and for key population groups. Poorer health outcomes universal to adolescents may reflect the complex multisystem developmental transition from childhood-to-adulthood, of which transition of healthcare services is a small part. The complex needs of this generation are well described and whilst examples of good practice are emerging, how best to support their transition to adulthood requires carefully tailored studies of cost-effective interventions that can be up scaled in resource limited settings.


Effect of the acute administration of high dose pentobarbital on human brain stem auditory and median nerve somatosensory evoked responses.
Brain stem auditory (BAERs) and median nerve somatosensory evoked responses (MnSSERs) were recorded from normal neural pathways during the induction of pentobarbital coma in six patients undergoing elective excision of complex arteriovenous malformations. Each patient received a 33-minute infusion of pentobarbital at a rate of 0.6 mg/kg (total dose, 19.8 mg/kg). This regimen resulted in burst suppression or isoelectricity of the electroencephalogram in all patients. Although statistically significant changes in latency and amplitude occurred, both BAERs and MnSSERs were readily recordable in all patients throughout the infusion. For the BAER, there were significant increases in the latencies of Waves III and V. However, these increases are sufficiently small that misinterpretation of these changes as an evolving neurological injury is unlikely, e.g., Wave V latency increased from 6.25 +/- 0.25 (SD) to 6.58 +/- 0.16 ms (P less than 0.006). For the MnSSER, changes of greater potential clinical relevance were observed. There were substantial increases in the latencies of the early components of the primary cortical response and in the central conduction time (e.g., CCT of 6.1 +/- 0.6 ms preinduction vs. 7.7 +/- 1.1 ms at t = 33 min, P less than 0.003), and the amplitude of the early cortical response (N20-P25) decreased by a mean of 45% (P less than 0.02). By contrast, subcortical components of the MnSSER (brachial plexus, upper cervical spine) were only minimally affected. We conclude that, in patients who are initially neurologically intact, BAERs and MnSSERs can be monitored effectively during pentobarbital coma and that the loss of these responses should not be ascribed to the effects of this drug alone. However, the interpretation of evoked response changes, particularly MnSSER changes, that occur during pentobarbital administration should take into account the dose-related changes in latency and amplitude that we have observed.


The structural basis of cysteine aminoacylation of tRNAPro by prolyl-tRNA synthetases.
Cysteinyl-tRNA synthetase is an essential enzyme required for protein synthesis. Genes encoding this protein have not been identified in Methanocaldococcus jannaschii, Methanothermobacter thermautotrophicus, or Methanopyrus kandleri. It has previously been proposed that the prolyl-tRNA synthetase (ProRS) enzymes in these organisms recognize either proline or cysteine and can aminoacylate their cognate tRNAs through a dual-specificity mechanism. We report five crystal structures at resolutions between 2.6 and 3.2 A: apo M. jannaschii ProRS, and M. thermautotrophicus ProRS in apo form and in complex with cysteinyl-sulfamoyl-, prolyl-sulfamoyl-, and alanyl-sulfamoyl-adenylates. These aminoacyl-adenylate analogues bind to a single active-site pocket and induce an identical set of conformational changes in loops around the active site when compared with the ligand-free conformation of ProRS. The cysteinyl- and prolyl-adenylate analogues have similar, nanomolar affinities for M. thermautotrophicus ProRS. Homology modeling of tRNA onto these adenylate complexes places the 3'-OH of A76 in an appropriate position for the transfer of any of the three amino acids to tRNA. Thus, these structures explain recent biochemical experiments showing that M. jannaschii ProRS misacylates tRNA(Pro) with cysteine, and argue against the proposal that these archaeal ProRS enzymes possess the dual capacity to aminoacylate both tRNA(Pro) and tRNA(Cys) with their cognate amino acids.


Gross anatomy of the muscle systems of Fasciola hepatica as visualized by phalloidin-fluorescence and confocal microscopy.
Neuropeptides, biogenic amines and acetylcholine are expressed abundantly within the nervous systems of parasitic flatworms, and are particularly evident in the innervation of the musculature. Such associations have implicated the nervous system in locomotion, host attachment and reproductive co-ordination. Information on the muscle systems of parasitic flatworms is generally sparse, in particular those muscles associated with the reproductive system, intestinal tract and attachment apparatus. Also, the use of sectioned material has left description of the 3-dimensional organization of the musculature largely unrecorded. Using fluorescein isothiocyanate (FITC)-labelled phalloidin as a site-specific probe for filamentous actin, applied to whole-mount preparations of adult Fasciola hepatica and examined by confocal scanning laser microscopy, the present work reports on the organization of the major muscle systems in this trematode parasite. A highly regular array of outer circular, intermediate longitudinal and inner diagonal fibres distinguishes the body wall musculature, which is also involved in the development of both ventral and oral suckers. Circular fibres dominate the duct walls of the male and female reproductive systems, whereas the muscles of the intestinal tract have a somewhat diffuse arrangement of fibres. An understanding of the structural complexity of the muscle systems of parasitic flatworms is considered as fundamental to the interpretation of results from physiological experiments.


Mitochondrial DNA Haplogroups and Delirium During Sepsis.
Studies suggest that mitochondrial dysfunction underlies some forms of sepsis-induced organ failure. We sought to test the hypothesis that variations in mitochondrial DNA haplogroup affect susceptibility to sepsis-associated delirium, a common manifestation of acute brain dysfunction during sepsis. Retrospective cohort study. Medical and surgical ICUs at a large tertiary care center. Caucasian and African American adults with sepsis. We determined each patient's mitochondrial DNA haplogroup using single-nucleotide polymorphisms genotyping data in a DNA databank and extracted outcomes from linked electronic medical records. We then used zero-inflated negative binomial regression to analyze age-adjusted associations between mitochondrial DNA haplogroups and duration of delirium, identified using the Confusion Assessment Method for the ICU. Eight-hundred ten patients accounted for 958 sepsis admissions, with 802 (84%) by Caucasians and 156 (16%) by African Americans. In total, 795 patient admissions (83%) involved one or more days of delirium. The 7% of Caucasians belonging to mitochondrial DNA haplogroup clade IWX experienced more delirium than the 49% in haplogroup H, the most common Caucasian haplogroup (age-adjusted rate ratio for delirium 1.36; 95% CI, 1.13-1.64; p = 0.001). Alternatively, among African Americans the 24% in haplogroup L2 experienced less delirium than those in haplogroup L3, the most common African haplogroup (adjusted rate ratio for delirium 0.60; 95% CI, 0.38-0.94; p = 0.03). Variations in mitochondrial DNA are associated with development of and protection from delirium in Caucasians and African Americans during sepsis. Future studies are now required to determine whether mitochondrial DNA and mitochondrial dysfunction contribute to the pathogenesis of delirium during sepsis so that targeted treatments can be developed.


[Value of determining serum cholesterol levels in patients with coronary heart disease].
Cholesterol synthesis (HMG-CoA reductase) inhibitors have proven their value in preventing cardiovascular events, especially in patients with manifest coronary heart disease. Besides cholesterol lowering a number of effects have been described which may contribute to the beneficial influence of these agents on the process of atherosclerosis. Measurement of serum lipids is still necessary for various reasons, namely, to know the degree of elevation in serum cholesterol and specific disturbances in lipid metabolism, the extent to which serum lipids must be lowered and the compliance with cholesterol lowering therapy.


[Alcohol pathology in a general hospital].
Screening of chronic alcohol intoxication (CAI) based on parallel use of test-questionnaires (CAGE, PAS-questionnaire) and assessment of organic evidence of alcohol abuse were employed in a clinicoepidemiological case-control survey covering 1191 patients aged 16-91 years admitted to a general hospital. 3335 autopsy protocols were analysed by pathoanatomical and forensic-medical criteria of CAI. It was found that CAI, directly or indirectly, raises risk of a wide spectrum of internal diseases which, according to ICD-10, are not directly related to alcohol. Each 5th hospitalization was caused by CAI sequela. Combined use of tests-questionnaires and consideration of physical stigms of CAI are proposed for screening of alcohol abuse and latent alcoholism in general medical practice. The authors think it necessary to establish addictological service at large general hospitals.


Molecular Mechanisms of Lipopolysaccharide (LPS) Induced Inflammation in an Immortalized Ovine Luteal Endothelial Cell Line (OLENDO).
Escherichia coli (E. coli) is the most common Gram-negative bacterium causing infection of the uterus or mammary gland and is one of the major causes of infertility in livestock. In those animals affected by E. coli driven LPS-mediated infections, fertility problems occur in part due to disrupted follicular and luteal functionality. However, the molecular mechanisms by which LPS induces inflammation, and specifically, the role of LPS in the disruption of capillary morphogenesis and endothelial barrier function remain unclear. Here, we hypothesized that LPS may lead to alterations in luteal angiogenesis and vascular function by inducing inflammatory reactions in endothelial cells. Accordingly, OLENDO cells were treated with LPS followed by evaluation of the expression of selected representative proinflammatory cytokines: NF-kB, IL6, IL8, TNFα, and ICAM 1. While TNFα was not affected by treatment with LPS, transcripts of NF-kB, IL6, and IL8 were affected in a dosage-dependent manner. Additionally, the activity of TLR2 and TLR4 was blocked, resulting in suppression of the LPS-induced expression of ICAM 1, NF-kB, IL6, and IL8. Inhibition of the PKA or MAPK/ERK pathways suppressed the LPS-stimulated expression of NF-kB, IL6, and IL8, whereas blocking the PKC pathway had the opposite effect. Furthermore, LPS-induced phosphorylation of Erk1 and Erk2 was inhibited when the TLR4 or MAPK/ERK pathways were blocked. Finally, LPS seems to induce inflammatory processes in OLENDO cells via TLR2 and TLR4, utilizing different signaling pathways.


Management of Enthesitis in Patients With Psoriatic Arthritis: An Updated Literature Review Informing the 2021 GRAPPA Treatment Recommendations.
Enthesitis is a key pathological and clinical feature of psoriatic arthritis (PsA) in children and adults. Enthesitis is typically assessed clinically using several validated enthesitis scoring systems that have been used in clinical trials. Enthesitis treatment response has been reported as change in the total enthesitis score or the proportion of patients who achieved complete resolution. The majority of trials in PsA did not require patients to have enthesitis at study entry since enthesitis was evaluated only as a secondary outcome. Despite the inherent limitations of the clinical assessment of enthesitis, imaging of the entheses using ultrasound or magnetic resonance imaging has rarely been used in clinical trials to assess response to treatment of enthesitis. This systematic review summarizes existing evidence regarding pharmaceutical and nonpharmaceutical interventions for enthesitis in patients with PsA to facilitate an evidence-based update of the Group for Research and Assessment in Psoriasis and Psoriatic Arthritis (GRAPPA) treatment recommendations for PsA. We performed a systematic literature review to identify 41 randomized clinical trials that reported enthesitis treatment response in patients with PsA. For each intervention, the response effect size was summarized and the quality of evidence was graded. Recommendations were then formulated for the various pharmacological and nonpharmacological therapies. We included 41 randomized clinical trials in our review and graded each intervention. Several classes of systemic conventional and advanced therapies and local measures were recommended for active enthesitis in patients with PsA.


Bone mineral density of the spine in normal Japanese subjects using dual-energy X-ray absorptiometry: effect of obesity and menopausal status.
Bone mineral density (BMD) of the lumbar spine was measured to determine normal Japanese values and to examine the effect of obesity and menopausal status on BMD. Normal Japanese subjects (N = 1,296, 1,048 women and 248 men) were examined using dual-energy X-ray absorptiometry. BMD for men peaked between age 20 and 29. For women, there was abrupt bone loss after age 50. Obese women within the same age bracket had a higher BMD than thin women after age 40-49. We determined that BMD began to decline during the irregular menstruation period before the onset of menopause. We conclude that there is a positive correlation between obesity and BMD, particularly in postmenopausal women. In addition, we found that bone loss related to menopause begins during the irregular menstruation period before menopause.


Glucose-Induced Transcriptional Hysteresis: Role in Obesity, Metabolic Memory, Diabetes, and Aging.
During differentiation transient, inducers produce permanent changes in gene expression. A similar phenomenon, transcriptional hysteresis, produced by transient or prolonged exposure to glucose, leads to cumulative, persistent, and largely irreversible effects on glucose-regulated gene expression, and may drive key aspects of metabolic memory, obesity, diabetes, and aging, and explain the protective effects of dietary restriction during aging. The most relevant effects of glucose-induced transcriptional hysteresis are the persistent effects of elevated glucose on genes that control glucose metabolism itself. A key observation is that, as with the lac operon, glucose induces genes that promote glycolysis and inhibits gene expression of alternative metabolic pathways including the pentose pathway, beta oxidation, and the TCA cycle. A similar pattern of metabolic gene expression is observed during aging, suggesting that cumulative exposure to glucose during aging produces this metabolic shift. Conversely, dietary restriction, which increases lifespan and delays age-related impairments, produces the opposite metabolic profile, leading to a shift away from glycolysis and toward the use of alternative substrates, including lipid and ketone metabolisms. The effect of glucose on gene expression leads to a positive feedback loop that leads to metastable persistent expression of genes that promote glycolysis and inhibit alternative pathways, a phenomenon first observed in the regulation of the lac operon. On the other hand, this pattern of gene expression can also be inhibited by activation of peroxisome proliferator activating receptor transcription factors that promote beta oxidation and inhibit metabolism of glucose-derived carbon bonds in the TCA cycle. Several pathological consequences may arise from glucose-induced transcriptional hysteresis. First, elevated glucose induces glycolytic genes in pancreatic beta cells, which induces a semi-stable persistent increase in insulin secretion, which could drive obesity and insulin resistance, and also due to glucose toxicity could eventually lead to beta-cell decompensation and diabetes. Diabetic complications persist even after complete normalization of glucose, a phenomenon known as metabolic memory. This too can be explained by persistent bistable expression of glucose-induced glycolytic genes.


Mammalian DNA polymerase alpha: a replication competent holoenzyme form from calf thymus.
A complex "replication competent" holoenzyme form of DNA polymerase alpha (RC-alpha) was purified 10,000 fold from calf thymus through the use of an assay employing primed single stranded circular DNA template. The RC-alpha form could partially replicate a double-stranded oligo(dT)-tailed linear DNA and could completely convert primed single-stranded circular DNA to its double stranded form. The RC-alpha was resolved by denaturing gel electrophoresis into at least 10 discrete polypeptide species ranging in apparent molecular mass from 200 to 47 kilodaltons; three of the bands (apparent Mr of 200, 118 and 63 kilodaltons) displayed DNA polymerase activity in denaturing gel activity assay. The isolation of RC-alpha required the use of absolutely fresh calf thymus, the inclusion of ATP and protease inhibitors throughout the purification procedure. Treatment of the RC-alpha with the neutralizing anti-DNA polymerase alpha monoclonal antibody SJK 132-20 (Tanaka et al. (1982), J. Biol. Chem. 257, 8386-8390) in nondenaturing conditions selected the complete set of 10 polypeptides, whereas treatment in denaturing conditions selected the 200 kilodalton catalytic DNA polymerase active polypeptide. The properties and the behaviour of the RC-alpha preparation following removal of specific polypeptides strongly suggested that the capacity of RC-alpha to extend and replicate long template requires the function of nonproteolysed form of the 200 kilodaltons catalytic DNA polymerase core and at least 6 other auxiliary polypeptides of, respectively, 98, 87, 63, 54, 49 and 47 kilodaltons.


Honey bee aggression supports a link between gene regulation and behavioral evolution.
A prominent theory states that animal phenotypes arise by evolutionary changes in gene regulation, but the extent to which this theory holds true for behavioral evolution is not known. Because "nature and nurture" are now understood to involve hereditary and environmental influences on gene expression, we studied whether environmental influences on a behavioral phenotype, i.e., aggression, could have evolved into inherited differences via changes in gene expression. Here, with microarray analysis of honey bees, we show that aggression-related genes with inherited patterns of brain expression are also environmentally regulated. There were expression differences in the brain for hundreds of genes between the highly aggressive Africanized honey bee compared with European honey bee (EHB) subspecies. Similar results were obtained for EHB in response to exposure to alarm pheromone (which provokes aggression) and when comparing old and young bees (aggressive tendencies increase with age). There was significant overlap of the gene lists generated from these three microarray experiments. Moreover, there was statistical enrichment of several of the same cis regulatory motifs in promoters of genes on all three gene lists. Aggression shows a remarkably robust brain molecular signature regardless of whether it occurs because of inherited, age-related, or environmental (social) factors. It appears that one element in the evolution of different degrees of aggressive behavior in honey bees involved changes in regulation of genes that mediate the response to alarm pheromone.


A comparison of the adhesion, coaggregation and cell-surface hydrophobicity properties of fibrillar and fimbriate strains of Streptococcus salivarius.
Fibrillar and fimbriate strains of Streptococcus salivarius were compared for their ability to adhere to buccal epithelial cells and saliva-coated hydroxyapatite beads, and for their ability to coaggregate with Veillonella strains. The fibrillar Lancefield group K strains adhered statistically significantly better to both buccal epithelial cells and saliva-coated hydroxyapatite beads than the fimbriate strains, which lacked the Lancefield group K antigen. After 1 h the fibrillar strains coaggregated statistically significantly better than the fimbriate strains with V. parvula strain V1, but after 24 h, coaggregation both of fibrillar and of fimbriate strains reached approximately 90%. Freshly isolated Veillonella strains all coaggregated with the S. salivarius strains, but the percentage coaggregation varied considerably after 1 h depending on the Veillonella strain. Coaggregation was independent of the presence of Ca2+. S. salivarius strain HB-V5, a mutant of strain HB that had lost the Veillonella-binding protein, coaggregated weakly with V. parvula strain V1, but coaggregated very well with other wild-type veillonellae, suggesting the presence of an alternative mechanism for Veillonella-binding for strain HB. Fibrillar strains were, therefore, more adhesive to oral surfaces and coaggregated with veillonellae after 1 h better than the fimbriate S. salivarius strains. Both fibrillar and fimbriate strains were highly hydrophobic in the hexadecane-buffer partition assay.


Glucosinolates of the only three Brassicales indigenous to French Polynesia.
The glucosinolate (GL) profiles in leaf and stem of Rorippa sarmentosa (G.Forst. ex DC.) J.F.Macbr., Lepidium bidentatum Montin var. bidentatum, and Capparis spinosa subsp. cordifolia (Lam.) Fici indigenous to French Polynesia were investigated for the first time using LC-MS analysis. In the present study, we have established the presence of 8 known GLs in R. sarmentosa: 4-(methylsulfinyl)butyl- (1), but-3-enyl- (2), 5-(methylsulfinyl)pentyl- (3), 6-(methylsulfinyl)hexyl- (4), indol-3-ylmethyl- (6), 2-phenylethyl- (7), 8-(methylsulfinyl)octyl- (8), and 9-(methylsulfinyl)nonyl- (9) GLs. We have also tentatively identified for the first time the presence in R. sarmentosa of 7-(methylsulfinyl)heptyl GL (5). In addition, we have identified two known GLs in L. bidentatum var. bidentatum: benzyl- (10) and 4-methoxybenzyl- (11) GLs. Finally, the known methyl GL (12) was shown to be largely predominant in C. spinosa subsp. cordifolia.


[Role of natriuresis in the prophylactic effect of adaptation to hypoxia in hereditary hypertension].
It was shown in experiments on male Wistar rats and rats with spontaneous hereditary hypertension (SHH) that the development of SHH was accompanied by a significant decrease of the renal excretion of water and sodium. The acute hypoxia (the rising in the barochamber to the high altitude of 4400 m during 6 hours) in Wistar rats resulted in a significant decrease of diuresis, glomerular filtration. K and (insignificantly) Na excretion; while in SHH rats acute hypoxia increased diuresis significantly and enhanced the Na and K excretion. The adaptation to hypoxia during 6 hours per day resulted in an increase of diuresis, glomerular filtration, Na excretion and the Na/K coefficient of urine in both the animal lines. During the first month of adaptation the changes were more pronounced in SHH than in Wistar rats, as a result the hypertension did not develop in SHH rats. The relationship of the hypotensive effect of the high altitude adaptation with its natriuretic effect was shown. It is concluded that the natriuretic effect is an important and, probably, the main mechanism of the prophylactic effect of the adaptation to hypoxia on the development of hereditary hypertension.


[Ocular symptoms associated with dupilumab in atopic dermatitis].
Dupilumab, the first registered biological for the treatment of atopic dermatitis, frequently causes adverse ocular events. These often take the form of mild conjunctivitis or blepharoconjunctivitis, however, more serious adverse ocular events have also been reported, including limbal stem cell insufficiency. A 56-year-old man was treated with dupilumab for severe atopic dermatitis. His eczema improved greatly, but he developed bilateral conjunctivitis with limbal stem cell insufficiency in his right eye. After discontinuation of dupilumab, the eye symptoms gradually disappeared. After three months no conjunctivitis or limbal stem cell insufficiency was observed on ophthalmological examination. Adverse ocular events are often seen in the treatment of atopic dermatitis with dupilumab. Caution is advised if severe adverse events occur. Therefore, it is important to identify the underlying pathophysiological mechanism of these adverse events.