Sustained induction of hepatocyte growth factor by sensitization with freeze-thawed hepatic tissue.
Hepatocyte growth factor (HGF) is a mitogen for hepatocytes, and has therapeutic potential for fibrotic/cirrhotic liver. Therefore, the induction of HGF in vivo is considered to be useful in the treatment of liver dysfunction caused by cirrhosis, chronic hepatitis, or extensive surgical resection. In this study, we examined the sustained induction of HGF by inoculation of freeze-thawed hepatic tissue (FTHT). Serum from rats inoculated with FTHT increased [3H]thymidine incorporation i.e., increased DNA synthesis, in primary cultured rat hepatocytes. The DNA synthesis was significantly promoted by the addition of the FTHT-sensitized serum, while this DNA synthesis was inhibited by neutralizing anti-rat HGF antibody. The concentration of HGF in the FTHT-sensitized serum was increased by day 3 after the inoculation. The time of HGF induction was dependent on the inoculated volume of FTHT, but peaks of HGF concentration were found on day 5 with different volumes of FTHT. Injurins, inducers of HGF, were also induced in the FTHT-sensitized rats, with their peak levels on day 3. The FTHT inoculated tissue showed inflammatory cell infiltration, which was gradually absorbed, and had completely disappeared by day 14 after the inoculation. Although mild inflammatory cell infiltration was observed in non-freeze-thawed inoculated hepatic tissue (NFHT) a tight capsule formed around the NFHT, and was scarcely phagocytized on day 14. These results suggest that FTHT inoculation induces HGF sustainedly through the increased synthesis of injurins, and that freeze-thawed tissue, which is easily phagocytized, is important for the sustained induction of HGF.


Spinal cord disorder due to spinal epidural abscess secondary to thoracic facet joint septic arthritis-a rare case with a surprising evolution.
Septic arthritis of a facet joint (SAFJ) is a relatively rare medical entity and the lumbar segment is its most frequent location. Although unusual, a spinal epidural abscess (SEA) can occur as a complication of SAFJ and possibly damage the spinal cord. A 53-year-old woman presented with acute right dorsal pain, fever, loss of sphincters control, and paraparesis without causal factor. Forty-eight hours after the symptoms started, imaging revealed a thoracic posterior epidural collection causing compression of the spinal cord and features suggestive of septic arthritis of right T5-T6 facet joint. She underwent an urgent laminectomy and drainage of the abscess. Both blood and abscess cultures isolated Staphylococcus aureus. A diagnosis of complete paraplegia grade A of the ASIA (American Spinal Injury Association) Impairment Scale (AIS) with neurologic level T10 was made 5 days after surgery. She took 3 months of an intensive rehabilitation program at our Physical and Rehabilitation Medicine ward. With an unexpected favorable evolution, both neurological and functional, she was discharged with an incomplete paraplegia AIS D and functionally independent in all tasks. This case reveals an infrequent medical condition on a rarely affected spinal segment. An early diagnosis and proper treatment of SAFJ and SEA are of major importance to avoid severe related consequences. Patients with spinal cord injury with severe neurological deficits due to these conditions greatly benefit from an interdisciplinary rehabilitation program to improve neuromotor and functional status.


COUP-TFI is a potential regulator of retinoic acid-modulated development in Xenopus embryos.
Isomers of retinoic acid are considered likely regulators of developmental pattern formation in vertebrate embryos. The orphan receptor COUP-TFI, which can alter cellular responses to retinoic acid in cultured cells, is expressed in distinct regions of the developing zebrafish and mouse anterior central nervous system. We asked if COUP-TFI can modulate retinoic acid signaling and anterior neural development in a vertebrate embryo by examining: (1) whether COUP-TFI could alter transcriptional responses to retinoic acid in Xenopus embryonic explants, and (2) whether misexpression of COUP-TFI could regulate anterior neural gene expression during early Xenopus development. The results from these studies show that COUP-TFI is a potent regulator of retinoic acid-induced gene expression in Xenopus embryonic cells, and that misexpression of COUP-TFI causes deficiencies in anterior neural structures and head development in Xenopus embryos with a concomitant change in anterior neural gene expression. These results support the proposition that COUP-TFI has a role in the elaboration and patterning of anterior neural gene expression in vertebrates, possibly via effects on the retinoic acid signaling pathways.


Enzyme histochemical localisation of alkaline phosphatase activity in osteogenesis imperfecta bone and growth plate: a preliminary study.
At the ultrastructural level alkaline phosphatase has been studied in calcifying cartilage but not in bone. The aim of this study was to assess if there is an osteoblast dysfunction in Osteogenesis Imperfecta (OI) with respect to alkaline phosphatase activity. Specimens from three OI type II foetal femoral bones, two OI type II growth plates, one normal foetal femoral bone and growth plate, one OI type III femoral bone specimen and one normal juvenile bone specimens were examined using modified lead nitrate method to identify alkaline phosphatase reactivity. The electron dense reaction product (indicative of the presence of alkaline phosphatase) was demonstrable on the cell membrane of the osteoblasts, as focal concentrations in the collagen osteoid and on the mineralisation front of normal bone. In normal bone the intensity of the reaction seemed to be stronger than in OI bone and appeared as a continuous black line along the osteoblast cell membranes. In OI bone the reaction product only appeared as a few electron dense beads along the osteoblast cell membrane. There appeared to be reduced and diffuse reaction product on OI osteoblasts, thus implying either a reduced level and/or altered activity of alkaline phosphatase and hence a dysfunction of osteoblasts. This confirms the findings of the previous report of the impaired activity of alkaline phosphatase in OI osteoblasts. Even in the OI growth plate, hypertrophic chondrocytes showed less intense reaction product than the chondrocytes in the normal growth plate. The normal human growth plates used in this study showed a similar pattern, but in the OI growth plate even the hypertrophic zone, where the alkaline phosphatase activity is reported to be high, showed less intense reaction product. Biochemical reports indicate that alkaline phosphatase levels are normal in cultured OI cell lines, yet ultrastructural histochemical observations reported here, show reduced enzyme localisation and this may suggest reduced amounts of protein or reduced activity at the tissue level.


Development and usability of a web-based patient-tailored tool to support adherence to urate-lowering therapy in gout.
The aim of this study is to develop and assess usability of a web-based patient-tailored tool to support adherence to urate-lowering therapy (ULT) among gout patients in a clinical setting. The content of the tool was based on the Integrated Change (I-Change) model. This model combines various socio-cognitive theories and assumes behavioral change is a result of becoming aware of the necessity of change by integrating pre-motivational, motivational, and post-motivational factors. An expert group (five gout experts, three health services researchers, and one health behavior expert) was assembled that decided in three meetings on the tool's specific content (assessments and personalized feedback) using information from preparatory qualitative studies and literature reviews. Usability was tested by a think aloud approach and validated usability questionnaires. The I-Change Gout tool contains three consecutive sessions comprising 80 questions, 66 tailored textual feedback messages, and 40 tailored animated videos. Navigation through the sessions was determined by the patients' intention to adapt suboptimal ULT adherence. After the sessions, patients receive an overview of the personalized advices and plans to support ULT adherence. Usability testing among 20 gout patients that (ever) used ULT and seven healthcare professionals revealed an overall score for the tool of 8.4 ± 0.9 and 7.7 ± 1.0 (scale 1-10). Furthermore, participants reported a high intention to use and/or recommend the tool to others. Participants identified some issues for further improvement (e.g. redundant questions, technical issues, and text readability). If relevant, these were subsequently implemented in the I-Change Gout tool, to allow further testing among the following participants. This study provides initial support for the usability by patients and healthcare professionals of the I-Change Gout tool to support ULT adherence behavior.


A type 2 (Th2-like) pattern of immune response predominates in the pulmonary interstitium of patients with cryptogenic fibrosing alveolitis (CFA).
CFA is an inflammatory condition of the lungs resulting in scarring, pulmonary failure and death. The etiology of the disease is unknown, but the pathogenesis is believed to involve a persistent immunological reaction to unidentified antigen in the lung resulting in tissue damage. Recent advances in our understanding of the immune system have shown that different patterns of stimulatory cytokines are produced at sites of inflammation by a range of cell types. Patterns of cytokine reproduction by inflammatory cells are recognized to be associated with different patterns of immunological response, and these have been described as type 1 (or Th1-like) and type 2 (or Th2-like) on this basis. We have studied cytokine expression in the intestinal inflammatory cell infiltrate in lung tissue from patients with CFA using mRNA in situ hybridization and immunohistochemistry. Our results show that while there is evidence for both a type 1 (characterized by interferon-gamma (IFN-gamma) and type 2 (characterized by IL-4 and IL-5) response present in CFA, the type 2 (or Th2) pattern of cytokines appears to predominate. This would be consistent with a possible role for the humoral immune response in the pathogenesis of this condition. In addition, recent evidence suggests that IL-4 and IFN-gamma may be important regulatory factors for pulmonary fibroblasts. The relative paucity of IFN-gamma may contribute to the excessive fibroblast activation, deposition of collagen and scar formation that occurs in CFA.


Retinal vascular profile in predicting incident cardiometabolic diseases among individuals with diabetes.
To determine the longitudinal associations between retinal vascular profile (RVP) and four major cardiometabolic diseases; and to quantify the predictive improvements when adding RVP beyond traditional risk factors in individuals with diabetes. Subjects were enrolled from the Singapore Epidemiology of Eye Disease (SEED) study, a multi-ethnic population-based cohort. Four incident cardiometabolic diseases, calculated over a ~ 6-year period, were considered: cardiovascular disease (CVD), hypertension (HTN), diabetic kidney disease (DKD), and hyperlipidemia (HLD). The RVP-vessel tortuosity, branching angle, branching coefficient, fractal dimension, vessel caliber, and DR status-was characterized at baseline using a computer-assisted program. Traditional risk factors at baseline included age, gender, ethnicity, smoking, blood pressure (BP), HbA1c, estimated glomerular filtration rate (eGFR), or cholesterol. The improvements in predictive performance when adding RVP (compared with only traditional risk factors) was calculated using several metrics including area under the receiver operating characteristics curve (AUC) and net reclassification improvement (NRI). Among 1770 individuals with diabetes, incidences were 6.3% (n = 79/1259) for CVD, 48.7% (n = 166/341) for HTN, 14.6% (n = 175/1199) for DKD, and 59.4% (n = 336/566) for HLD. DR preceded the onset of CVD (RR 1.85[1.14;3.00]) and DKD (1.44 [1.06;1.96]). Narrower arteriolar caliber preceding the onset of HTN (0.84 [0.72;0.99]), and changes in arteriolar branching angle preceded the onset of CVD (0.78 [0.62;0.98]) and HTN (1.15 [1.03;1.29]). The largest predictive improvement was found for HTN with AUC increment of 3.4% (p = .027) and better reclassification of 11.4% of the cases and 4.6% of the controls (p = .008). We found that RVPs improved the prediction of HTN in individuals with diabetes, but add limited information for CVD, DKD, and HLD predictions.


Involvement of pre- and postsynaptic receptors in catecholaminergic control of paradoxical sleep in man.
In an attempt to clarify the roles of pre- and postsynaptic catecholaminergic (CA) receptors in the regulation of paradoxical sleep (PS) in man, the effects of various doses of chlorpromazine (CPZ) and clonidine (CLN) have been investigated. The action of CPZ was biphasic, small doses enhancing and larger doses depressing the production of PS. However, the effect of CLN was monophasic, showing no modification after small doses, and a decrease in production of PS after moderate doses. In addition, a small dose of CLN, ineffective by itself, completely abolished the effect of a small dose of CPZ. These observations can be explained by preferential blockade of presynaptic alpha-receptors by a small dose of CPZ. They show, too, that it is possible to investigate in man the functional consequences of pharmacological manipulation of pre-synaptic receptors on cerebral CA neurons.


Clinical trial: single- and multiple-dose pharmacokinetics of polyethylene glycol (PEG-3350) in healthy young and elderly subjects.
The pharmacokinetics of polyethylene glycol 3350 (PEG-3350) have not been fully described because of lack of a sufficiently sensitive analytical method. To describe the pharmacokinetics of PEG-3350 in humans. A highly sensitive, high performance liquid chromatography with mass spectrometry (HPLC/MS/MS) method was developed for PEG-3350 in urine, plasma and faeces with quantification limits of 30 ng/mL, 100 ng/mL and 500 microg/g respectively. Noncompartmental pharmacokinetics methods were used and the effects of gender, age, renal status and dosing frequency were examined after the oral administration of 17 g to healthy volunteers. Peak PEG-3350 plasma concentrations occurred at 2-4 h and declined to nonquantifiable levels usually within 18 h after single and multiple doses, with a half-life of about 4-6 h. Steady state was reached within 5 days of dosing. Mean urinary excretion of the administered dose ranged from 0.19% to 0.25%. Age, gender or mild kidney impairment did not alter the pharmacokinetics of PEG-3350. Mean faecal excretion of the administered dose was 93% in young subjects. For the first time, a highly sensitive assay allowed comprehensive pharmacokinetics studies of PEG-3350 in humans. These studies confirmed that orally administered PEG-3350 is minimally absorbed, rapidly excreted and primarily eliminated via faeces.


Antibacterial activities of fluorescent nano assembled triphenylamine phosphonium ionic liquids.
Staphylococcus aureus, a Gram positive coccal bacterium is a major cause of nosocomial infection. We report the synthesis of new triphenylamine phosphonium ionic liquids which are able to self-assemble into multiwall nanoassemblies and to reveal a strong bactericidal activity (MIC=0.5mg/L) for Gram positive bacteria (including resistant strains) comparable to that of standard antibiotics. Time kill, metabolism and fluorescence confocal microscopy studies show a quasi-instantaneously penetration of the nanoassemblies inside the bacteria resulting of a rapid blocking (30min) of their proliferation. As confirmed by rezasurin reduction monitoring, these compounds strongly affect the bacterial metabolism and a Gram positive versus Gram negative selectivity is clearly observed. These fluorescent phosphonium ionic liquid might constitute a useful tool for both translocation studies and to tackle infectious diseases related to the field of implantology.


[Increased levels of cesium-137 and cesium-134 in 34 mushroom species following the reactor accident at Chernobyl].
From September to December of 1986, 85 samples of 34 species of wild mushrooms were collected, mostly in northern Baden and occasionally in neighboring areas. Cs-137, Cs-134, and the natural nuclide K-40 were determined by gamma-spectrometry. Samples from the Rhine Valley and the Kraichgau were generally much less contaminated than those from the Black Forest. With one exception, their radiocesium levels were below the Common Market limit of 600 Bq/kg. The exception was a sample of Laccaria amethystina with 1710 Bq/kg Cs-137 and 766 Bq/kg Cs-134. Relatively high levels were also found in Xerocomus badius, X. chrysenteron and Cantharellus tubaeformis. The hymenophore contained higher concentrations of radiocesium than the other parts of the body of the fruit. No correlation between the concentration of K-40 and that of the Cs nuclides was found. The ratio of Cs-137:Cs-134 was usually below 3 but reached the value of 11.8 in Piptoporus betulinus growing on birch wood. In this case, the substrate must have contained more radiocesium from the time before the Chernobyl reactor accident than from the time thereafter.


Therapeutic efficacy and safety of botulinum toxin type A in trigeminal neuralgia: a systematic review.
Trigeminal neuralgia is a common disorder caused mainly by compression of the trigeminal nerve root by an overlying blood vessel. Pharmacotherapy and surgery are ineffective or unsuitable in many patients. Therefore, other therapeutic modalities have been tried, including injection of botulinum toxin type A (BTX-A). This study aims to systematically review the therapeutic efficacy and safety of BTX-A in trigeminal neuralgia. PubMed, EMBASE, Cochrane Library Clinical Trials and Web of Science from January 1966 to March 2013 were searched with the terms of "botulinum toxin" AND "trigeminal neuralgia", and references of related articles were traced. Data on the efficacy and safety of BTX-A in this disorder were extracted and analyzed by at least 2 reviewers. Data for individual studies were reported, and pooled data were analyzed if appropriate. Five prospective studies and one double-blind, randomized, placebo-controlled study were identified. Response was achieved in approximately 70-100% of patients, and the mean pain intensity and frequency were reduced by approximately 60-100% at 4 weeks after treatment in most studies. Major adverse events were not reported. Available studies show BTX-A may be effective in treatment of trigeminal neuralgia. However, well-designed randomized, controlled, double-blinded trial is still lacking. Future BTX-A treatment studies on optimal dose, duration of the therapeutic efficacy, common AEs, and the time and indications for repeat injection would be promising.


Protective Effect of Breastfeeding Against Febrile Seizure: A Nationwide Study in Korea.
Breastfeeding is known to protect against febrile seizure (FS). Whether its impact continues throughout the childhood period is still controversial. Our objective was to investigate the protective effect of breastfeeding against FS stratified by age. We included children who participated in the National Health Screening Program for Infants and Children (NHSPIC) aged between four and six months between 2008 and 2014. Feeding type was confirmed based on the NHSPIC questionnaire, and data from the Korean National Health Insurance Service were used to determine FS cases during a five-year follow-up period. Among the 1,791,335 children, the most prevalent feeding type was exclusive breastfeeding (EB) (42.3%). FS occurred most frequently in the exclusive formula feeding (EF) group (12.2%), followed by the partial breastfeeding (PB) (11.3%) and EB groups (10.7%). Compared with the EF group, the adjusted odds ratio for FS was 0.87 (95% confidence interval, 0.86 to 0.88, P < 0.001) and 0.93 (0.92 to 0.94, P < 0.001) in the EB and PB groups, respectively. The protective effect by age 2.5 years was significant in both the EB (0.85; 0.84 to 0.86, P < 0.001) and PB (0.92; 0.90 to 0.93, P < 0.001) groups. In contrast, the protective effect was not significant in the PB group and inconsistent in the EB group after 2.5 years. Breastfeeding has a protective effect against FS in the most prevalent age period, from 0 to 2.5 years. Despite the limited effect after age 2.5 years, we support the current recommendation for prolonged breastfeeding to promote childhood health.


Effects of the Atrial Fibrillation Better Care Pathway on Outcomes Among Clinically Complex Chinese Patients With Atrial Fibrillation With Multimorbidity and Polypharmacy: A Report From the ChiOTEAF Registry.
Background Patients with atrial fibrillation commonly have complex clinical backgrounds of multimorbidity and polypharmacy. The Atrial Fibrillation Better Care (ABC) pathway has been developed to help deliver integrated and holistic care for patients with atrial fibrillation. In this ancillary analysis, we assessed the adherence to and the effectiveness of the ABC pathway at reducing adverse outcomes in Chinese patients with atrial fibrillation with a complex clinical background of multimorbidity or polypharmacy. Methods and Results The ChiOTEAF (Optimal Thromboprophylaxis in Elderly Chinese Patients With Atrial Fibrillation) registry is a prospective, multicenter, nationwide study conducted from October 2014 to December 2018. The primary outcomes of interest were the composite end point of all-cause death and thromboembolic events, as well as individual end points of all-cause death, thromboembolic events, and major bleeding. Multimorbidity was defined as the presence of ≥2 comorbidities, and polypharmacy was defined as the concomitant use of ≥5 medications. The eligible cohort included 4644 patients with multimorbidity, of whom 2610 (56.2%) had available data to assess the ABC pathway usage (mean age, 74.4±10.2; 42.8% women). Among patients with polypharmacy (n=2262; mean age, 74.6±10.1; 43.3% women), 1328 (58.7%) had available data to assess the use of the ABC pathway. Adherence to the ABC pathway was associated with a lower risk of the primary composite outcome among patients with multimorbidity (odds ratio, 0.48; 95% CI, 0.29-0.79) and in the polypharmacy group (odds ratio, 0.39; 95% CI, 0.19-0.78). Health-related quality of life was lower in the non-ABC-adherent group compared with the ABC-treated patients. Conclusions This nationwide real-world registry shows that adherence to the ABC pathway is associated with improved clinical outcomes and health-related quality of life in clinically complex Chinese patients with atrial fibrillation with multimorbidity or polypharmacy.


Anatomy and Aesthetics of the Labia Minora: The Ideal Vulva?
Female genital cosmetic surgery is becoming more and more widespread both in the field of plastic and gynaecological surgery. The increased demand for vulvar surgery is spurred by the belief that the vulva is abnormal in appearance. What is normal in terms of labial anatomy? Labia minora enlargement or hypertrophy remains a clinical diagnosis which is poorly defined as it could be considered a variation of the normal anatomy. Enlarged labia minora can cause functional, aesthetic and psychosocial problems. In reality, given the wide variety of vulvar morphology among people, it is a very subjective issue to define the "normal" vulva. The spread of nudity in the general media plays a major role in creating an artificial image and standards with regard to the ideal form. Physicians should be aware that the patient's self-perception of the normal or ideal vulva is highly influenced by the arguably distorted image related to our socio-psychological environment, as presented to us by the general media and internet. As physicians, we have to educate our patients on the variation of vulvar anatomy and the potential risks of these surgeries. Level of Evidence V This journal requires that authors assign a level of evidence to each article. For a full description of these evidence-based medicine ratings, please refer to Table of Contents or the online Instructions to Authors www.springer.com/00266 .


IRAP inhibition using HFI419 prevents moderate to severe acetylcholine mediated vasoconstriction in a rabbit model.
Coronary artery vasospasm (constriction) caused by reduced nitric oxide bioavailability leads to myocardial infarction. Reduced endothelial release of nitric oxide by the neurotransmitter acetylcholine, leads to paradoxical vasoconstriction as it binds to smooth muscle cell M3 receptors. Thus, inhibition of coronary artery vasospasm will improve clinical outcomes. Inhibition of insulin regulated aminopeptidase has been shown to improve vessel function, thus we tested the hypothesis that HFI419, an inhibitor of insulin regulated aminopeptidase, could reduce blood vessel constriction to acetylcholine. The abdominal aorta was excised from New Zealand white rabbits (n=15) and incubated with 3mM Hcy to induce vascular dysfunction in vitro for 1h. HFI419 was added 5min prior to assessment of vascular function by cumulative doses of acetylcholine. In some rings, vasoconstriction to acetylcholine was observed in aortic rings after pre-incubation with 3mM homocysteine. Incubation with HFI419 inhibited the vasoconstrictive response to acetylcholine, thus improving, but not normalizing, vascular function (11.5±8.9% relaxation vs 79.2±37% constriction, p<0.05). Similarly, in another group with mild vasoconstriction, HFI419 inhibited this effect (34.9±4.6% relaxation vs 11.1±5.2%, constriction, p<0.05). HFI419 had no effect on control aorta or aorta with mild aortic dysfunction. The present study shows that HFI419 prevents acetylcholine mediated vasoconstriction in dysfunctional blood vessels. HFI419 had no effect on normal vasodilation. Our results indicate a therapeutic potential of HFI419 in reducing coronary artery vasospasm.


Correlation of echocardiographic epicardial fat thickness with severity of coronary artery disease-an observational study.
Epicardial fat is an indirect contact with coronary arteries. There are some studies about the relationship between this fat and metabolic syndrome and it has considered as an indicator of cardiovascular risk. Several studies have addressed the association between epicardial fat thickness (EFT) and coronary artery disease (CAD) with conflicting results. The aim of our study was to evaluate the hypothesis that echocardiographic EFT thickness could be a marker severe CAD. Overall, 315 cases who underwent coronary angiography were classified in two groups: Normal and CAD. Measurement of EFT was done with echocardiography. The difference between mean EFT in two groups was analyzed. Califf scoring considered for severity of CAD. Then the relationship between EFT and age, sex, body mass index (BMI), serum lipids and severity of CAD was evaluated. The obtained data were compared by using ANCOVA test, Pearson and Spearman's partial correlation analyses. The EFT in CAD group was significantly higher than in normal group (5.4 ± 1.9 mm vs 4.4 ± 1.8 mm, p=0.0001). EFT had a positive relationship with Califf scoring of diseased coronary arteries (r=0.158 p=0.04), low-density lipoprotein cholesterol (p=0.04), female gender (p=0.02), BMI (p=0.001) and serum triglyceride levels (p=0.04). This study shows an association between EFT thickness and severity of CAD.


Cryopreservation of peripheral blood mononuclear cells using uncontrolled rate freezing.
Peripheral blood mononuclear cells are widely used as source material for anticancer immunotherapies. The conventional cryopreservation method for peripheral blood mononuclear cells is time-consuming and expansive, which involves controlled rate freezing followed by storage in liquid nitrogen. Instead, the convenient uncontrolled rate freezing cryopreservation method had been reported successfully in peripheral blood hematopoietic stem cells and peripheral blood progenitor cells. Therefore, we hypothesized that uncontrolled rate freezing cooling method maybe also applied to peripheral blood mononuclear cells cryopreservation. In this study, we evaluated the performance of uncontrolled rate freezing and controlled rate freezing cooling methods through cell recovery rate, viability, differentiation potential into cytokine-induced killer cells and the cellular properties of the cultured cytokine-induced killer cells. The results showed similar post-thaw viability and recovery rate in both controlled rate freezing and uncontrolled rate freezing cryopreserved peripheral blood mononuclear cells. Importantly, the uncontrolled rate freezing cryopreserved peripheral blood mononuclear cells exhibited higher growth ratio and earlier cell clustering during ex-vivo cytokine-induced killer cell culture than the controlled rate freezing ones. These two groups of expanded cytokine-induced killer cells also exhibited similar effector cell subset ratio and tumoricidal activity. In general, the performance of cryopreserved peripheral blood mononuclear cells using uncontrolled rate freezing cooling method, with the commercial cryoprotective agent CellBanker 2, was equal or better than the controlled rate freezing method. Our study implied that the combined use of cryoprotective agent CellBanker 2 and uncontrolled rate freezing could be a convenient cryopreservation method for peripheral blood mononuclear cells.


Endoscopic resection of adenoma-like mass in chronic ulcerative colitis using a combined endoscopic mucosal resection and cap assisted submucosal dissection technique.
Resection of an adenoma-like mass (ALM) in chronic ulcerative colitis (CUC) complicated by mucosal fibrosis has historically not been technically feasible. Endoscopic submucosal dissection techniques may now provide a therapeutic tool enabling the division of submucosal fibrotic scarring, hence enabling endoluminal resection for the first time in this select patient group. The aim was prospective evaluation of endoscopic submucosal dissection-assisted (ESD) resection of flat, sessile, and lateral spreading tumors in CUC complicated by submucosal desmoplasis. Clinical endpoints were postresection recurrence rates, R0 resection status, and complications. ESD-assisted endoscopic mucosal resection (EMR) using the Olympus KD-630L insulation-tipped knife was performed on selected lesions. Sixty-nine patients met inclusion criteria, of which 2 were excluded due to follow-up default. En bloc resection was performed in 52/67 (78%) cases with 15/67 (7%) requiring a piecemeal approach. R0 resection was achieved in 49/52 (94%) of lesions undergoing en bloc resection (perforation rate 2/67 [3%]). Bleeding complications occurred in 7/67 (10%) of cases. No metachronous circumscribed intraepithelial neoplastic lesions or cancer was detected at follow-up. At a median of 18 months follow-up, overall cure rates for the ESD-assisted EMR cohort was 66/67 (98%). We have shown for the first time that endoscopic resection of ALM even in the presence of complicating mucosal fibrosis is technically achievable using a combined ESD-assisted EMR technique. In an appropriately selected cohort, this technique may provide a technically feasible and clinically acceptable therapy where otherwise colectomy would be required.


No correlation of substantia nigra echogenicity and nigrostriatal degradation in Parkinson's disease.
Substantia nigra hyperechogenicity assessed by transcranial sonography is a typical finding in up to 90% of patients with idiopathic Parkinson's disease, although its value as a surrogate marker for disease progression in Parkinson's disease is controversial. (123) I-FP-CIT-single photon emission computed tomography (SPECT) represents an established paraclinical surrogate marker to quantify the nigrostriatal dopaminergic deficit in Parkinson's disease. Whereas most studies found no correlation between extent of substantia nigra echogenicity and the putaminal FP-CIT binding ratio, a more recent analysis reported opposite results. In 92 patients with Parkinson's disease the substantia nigra echogenicity was compared with the putaminal FP-CIT binding ratio using an investigator-independent SPECT analysis protocol and with several clinical parameters. No correlation was found between the substantia nigra hyperechogenicity and the FP-CIT binding ratio or the disease severity. Substantia nigra hyperechogenicity does not reflect the degree of the nigrostriatal degeneration or the clinical state of the disease progression.


Proton beam therapy for hepatocellular carcinoma with bile duct invasion.
Hepatocellular carcinoma (HCC) with bile duct invasion (BDI) (BDIHCC) has a poor prognosis. Moreover, due to the paucity of reports, there is no consensus regarding optimal management of this clinical condition yet. The aim of this study was to clarify the efficacy and safety of proton beam therapy (PBT) for BDIHCC. Between 2009 and 2018, 15 patients with BDIHCC underwent PBT at our institution. The overall survival (OS), local control (LC), and progression-free survival (PFS) curves were constructed using the Kaplan-Meier method. Toxicities were assessed using the Common Terminology Criteria of Adverse Events version 4.0. The median follow-up time was 23.4 months (range, 7.9-54.3). The median age was 71 years (range, 58-90 years). Many patients were Child A (n = 8, 53.3%) and most had solitary tumors (n = 11, 73.3%). Additionally, most patients had central type BDI (n = 11, 73%). The median tumor size was 4.0 cm (range, 1.5-8.0 cm). The 1-, 2-, and 3-year OS rates were 80.0%, 58.7% and 40.2%, respectively, and the corresponding LC and PFS rates were 93.3%, 93.3%, and 74.7% and 72.7%, 9.7%, and 0.0%, respectively. Acute grade 1/2 dermatitis (n = 7, 46.7%), and grades 2 (n = 1, 6.7%) and 3 (n = 1, 6.7%) cholangitis were observed. Late toxicities such as grade 3 gastric hemorrhage and pleural effusion were observed. No toxicities of grade 4 or higher were observed. PBT was feasible with tolerable toxicities for the treatment of BDIHCC.


Association of changes in self-reported vision and hearing impairments with depressive symptoms in middle-aged and older adults: Evidence from a nationwide longitudinal study in China.
This study assessed the longitudinal relationship between changes in self-reported vision impairment (VI) and hearing impairment (HI), and depressive symptoms in adults aged ≥45 years. Data from the China Health and Retirement Longitudinal Study for 2015 and 2018 were used, with a sample size of 10,050. VI and HI were self-reported. The 10-item Center for Epidemiologic Studies Depression Scale (CES-D10) was used to assess depressive symptoms. Total scores and clinically significant depressive symptoms (CES-D10 scores ≥ 10) were used as outcomes and analyzed using generalized estimating equations with identity link and logit link functions. Of all changes in HI with good vision, only persistent HI was associated with higher CES-D10 scores (OR 95% CI: 1.09-3.30). Both new-onset and persistent VI with good hearing were associated with CES-D10 scores (OR 95% CI, new-onset: 1.70-3.52; persistent: 1.58-2.78) and clinically significant depressive symptoms (OR 95% CI, new-onset: 1.41-2.56; persistent: 1.36-2.27). Persistent dual sensory impairment (DSI) was associated with the highest CES-D10 scores (OR 95% CI: 3.63-5.97) and the highest risk of clinically significant depressive symptoms (OR 95% CI: 1.78-2.85). Those who self-reported improvements in vision and hearing still had higher CES-D10 scores (OR 95% CI: 1.35-2.83) and a higher risk of clinically significant depressive symptoms (OR 95% CI: 1.03-2.02) than those maintaining good vision and hearing. Self-reported VI and DSI are closely associated with depressive symptoms. Because VI and DSI are modifiable, interventions for them could also prevent depressive symptoms.


Epidemiology of soccer-related head injury in children 5-14 years in Victoria, Australia.
Our aim was to use epidemiological data to determine the incidence of soccer-related head injuries in children aged 5-14 years who presented at emergency departments (EDs) or were admitted in hospitals in Victoria, Australia. ED presentation and hospital admission de-identified aggregate data were from the Victorian Injury Surveillance Unit. Soccer participation data were compared with the soccer-related head injury data to determine the incidence of this injury among these children. The incidence of ED presentations was 0.17% of children participating in soccer during the study period (financial years 2011-2012 to 2015-2016). The 10-14-years age group presented with more head injuries than the 5-9-years age group. For the admissions data, soccer had a significantly lower (P = 0.0379) incidence of head injury when compared with 'sport as a whole'. The low incidence of soccer-related head injuries presenting to an ED or admission to hospital is consistent with international findings.


[Saturday night retinopathy: A French case of post-compressive CRAO].
We report herein the first French case of Saturday Night Retinopathy. A 39-year-old man presented to the emergency room with unilateral vision loss in the left eye with redness but no pain. Visual acuity OS was "light perception" and OD 20/20. The left eye was hyperemic with a fixed, dilated pupil; fundus examination revealed a macular cherry-red spot within a pale, ischemic retina. The patient was admitted to a stroke centre. The neurological work-up and head CT were normal. He also underwent evaluation for possible carotid or cardiac etiologies, all of which were negative. Current smoking was the only cardiovascular risk factor found. The patient reportedly fell asleep face down at his kitchen table after consuming a large amount of alcohol, with his left eye pressed into his arm throughout the night. Six similar cases have been reported in the literature since 1973. The prognosis for vision is dismal. Only public awareness and prevention might avoid this serious functional disability.


Models for investigating the aetiology of cerebral arterial spasm: comparative responses of the human basilar artery with rat colon, anococcygeus, stomach fundus, and aorta and guinea-pig ileum and colon.
1 The pharmacological similarity between human basilar artery and a number of isolated tissues (rat colon, anococcygeus, stomach fundus and aorta and guinea-pig ileum and colon) has been assessed during investigations of the aetiology of cerebral arterial spasm. 2 The responses of each of the six tissues to human normal and abnormal cerebrospinal fluid (CSF) and to human serum were compared with those of the human isolated basilar artery. 3 These studies revealed the presence of a vasodilator factor in CSF from subarachnoid haemorrhage patients and several tissues may be of use for further work in identifying the vasoactive substances in CSF. 4 No tissue displayed exactly the same spectrum of biological reactivity as the human basilar artery. The rat stomach fundus showed the closest similarity and was further studied for similarities in drug-induced responses. 5 The rat stomach fundus, like the human basilar artery, was contracted by 5-hydroxytryptamine, prostaglandin F2alpha and histamine and relaxed by dopamine. However, noradrenaline relaxed the fundus but contracted the basilar artery.


Pseudoaneurysm in the iliac fossa after renal transplantation--treatment with ultrasound-guided thrombin injection.
We report the case of a 57-year-old female who developed a large pseudoaneurysm of the right circumflex iliac artery 15 months after renal transplantation. Minimal invasive treatment was successfully performed with ultrasound-guided thrombin injection using the B-flow mode.


Ventilation strategies in paediatric inhalation injury.
Inhalation injury increases morbidity and mortality in burns victims. While the diagnosis remains largely clinical, bronchoscopy is also helpful to diagnose and grade the severity of any injury. Inhalation injury results from direct thermal injury or chemical irritation of the respiratory tract, systemic toxicity from inhaled substances, or a combination of these factors. While endotracheal intubation is essential in cases where upper airway obstruction may occur, it has its own risks and should not be performed prophylactically in all cases of inhalation injury. The evidence-base informing the selection of optimal ventilation strategy in inhalation injury is sparse, and most recommendations are based on extrapolation from (largely adult) studies in acute respiratory distress syndrome (ARDS). Conventional ventilation using a lung-protective approach (i.e. low tidal volume, limited plateau pressure, and permissive hypercarbia) is recommended as the initial approach if invasive ventilation is required; various rescue strategies may become necessary if there is a poor response. The efficacy of many widely used pharmacologic adjuncts in inhalation injury remains uncertain. Further research is urgently required to address these gaps in our knowledge.


Multiplexed Thiol Reactivity Profiling for Target Discovery of Electrophilic Natural Products.
Electrophilic groups, such as Michael acceptors, expoxides, are common motifs in natural products (NPs). Electrophilic NPs can act through covalent modification of cysteinyl thiols on functional proteins, and exhibit potent cytotoxicity and anti-inflammatory/cancer activities. Here we describe a new chemoproteomic strategy, termed multiplexed thiol reactivity profiling (MTRP), and its use in target discovery of electrophilic NPs. We demonstrate the utility of MTRP by identifying cellular targets of gambogic acid, an electrophilic NP that is currently under evaluation in clinical trials as anticancer agent. Moreover, MTRP enables simultaneous comparison of seven structurally diversified α,β-unsaturated γ-lactones, which provides insights into the relative proteomic reactivity and target preference of diverse structural scaffolds coupled to a common electrophilic motif and reveals various potential druggable targets with liganded cysteines. We anticipate that this new method for thiol reactivity profiling in a multiplexed manner will find broad application in redox biology and drug discovery.


Engineering a zinc binding site into the de novo designed protein DS119 with a βαβ structure.
Functional proteins designed de novo have potential application in chemical engineering, agriculture and healthcare. Metal binding sites are commonly used to incorporate functions. Based on a de novo designed protein DS119 with a βαβ structure, we have computationally engineered zinc binding sites into it using a home-made searching program. Seven out of the eight designed sequences tested were shown to bind Zn(2+) with micromolar affinity, and one of them bound Zn(2+) with 1:1 stoichiometry. This is the first time that metalloproteins with an α, β mixed structure have been designed from scratch.


The effect of alcohol on blood glucose in Type 1 diabetes--metabolic modelling and integration in a decision support system.
We have recently shown, in studies with patients with Type 1 (insulin dependent) diabetes, that alcohol intake at 21:00 h significantly reduced blood glucose values after 10-12 h, compared with control studies with no alcohol. We hypothesised that this was due to the following effects of alcohol: (1) alcohol metabolism increases NADH, leading to a reduction in hepatic gluconeogenesis; (2) increased glycogen phosphorylase activity depletes hepatic glycogen stores; (3) after the alcohol is metabolised, hepatic insulin sensitivity is increased, leading to the restoration of glycogen stores and reduction in blood glucose levels; and (4) consequently, after several hours, glycogen stores and insulin sensitivity return to normal. A model describing these changes (DiasNet-Alcohol) was implemented into the DiasNet model of human glucose metabolism. Our study suggests that the DiasNet-Alcohol model gives a reasonable approximation of these effects of alcohol on blood glucose concentration observed in our study and supports our hypothesis for the mechanism behind these effects in Type 1 diabetes.


A quantitative method for the simultaneous detection of SR9009 and SR9011 in equine plasma using liquid chromatography-electrospray ionization-tandem mass spectrometry.
SR9009 and SR9011 are metabolic modulators pharmacologically targeting REV-ERB receptors as synthetic agonists. A liquid chromatography-tandem mass spectrometry method for the detection of SR9009 and SR9011 in equine plasma was developed and validated. Plasma samples were pretreated by protein precipitation with methanol and were loaded onto an ACQUITY ultra performance liquid chromatography high-strength silica C18 column (2.1 × 150 mm, 1.8 μm) for chromatographic separation. The mobile phase consisted of 5-mM ammonium formate (pH 3.0) in distilled water and 0.1% formic acid in acetonitrile, and a gradient elution was used at a flow rate of 0.25 ml/min. For the mass spectrometry detection, the selected reaction monitoring mode was used with transitions of 438.2 → 124.9 for SR9009, 479.2 → 125.1 for SR9011, and 292.2 → 109.1 for the internal standard (testosterone-d3) in the positive ionization mode. The linearity, lower limit of quantification, intra- and inter-day precision, accuracy, matrix effect, recovery, and stability were evaluated. The method was found to be accurate and reproducible for the quantitation of SR9009 and SR9011. The developed method was successfully applied to plasma samples of thoroughbreds injected intramuscularly with SR9009.


A simplified Timm stain procedure compatible with formaldehyde fixation and routine paraffin embedding of rat brain.
A simplified sulphide/silver technique for visualizing transitional metals in rat brain is described. It consists of: (1) sulphide perfusion before formaldehyde perfusion, (2) routine paraffin embedding and sectioning, and (3) silver staining with physical developer. This method is compatible with formaldehyde fixation, eliminates the need for post-fixation in Carnoy's solution before embedding and does not require low paraffin temperatures during embedding. Once embedded, brains can be stored for months before sectioning without loss of stain quality.


Vaccination against foot-and-mouth disease virus confers complete clinical protection in 7 days and partial protection in 4 days: Use in emergency outbreak response.
Recent outbreaks of foot-and-mouth disease virus (FMDV) demonstrate that this highly contagious viral infection of cloven hoofed animals continues to be a significant economic problem worldwide. Debate about the most effective way to respond to outbreaks of FMDV in disease free countries continues to center on the use of vaccines. In this report, we present data showing that a commercially available, standard dose vaccine formulation can fully protect cattle against direct challenge with the virus in as little as 7 days with no carrier transmission to naïve animals. Cattle challenged 4 days after vaccination have reduced disease severity, no detectable virus in blood and little virus shedding from nasal secretions. These significant effects at 4 days post vaccination, confirmed in two separate trials, support the value of using currently available vaccines as a first line of defense against foot-and-mouth disease (FMD) outbreaks.


Eugenol relieves the pathological manifestations of Alzheimer's disease in 5×FAD mice.
Alzheimer's disease (AD) is a progressive neurodegenerative disease characterized by the accumulation of amyloid-β (Aβ) and excessive neuroinflammation, resulting in neuronal cell death and cognitive impairments. Eugenol, a phenylpropene, is the main component of Syzygium aromaticum L. (Myrtaceae) and has multiple therapeutic effects, including neuroprotective and anti-inflammatory effects, through multimodal mechanisms. We aimed to investigate the effect of eugenol on AD pathologies using a 5× familiar AD (5×FAD) mouse model. Eight-month-old 5×FAD and wild-type mice were administered with eugenol (10 or 30 mg/kg/day, p.o) for 2 months. Y-maze and Morris water maze tests were performed to assess the cognitive function of mice. After the behavioral test, molecular analysis was conducted to investigate the therapeutic mechanism of eugenol. Our findings indicate that eugenol treatment effectively mitigated cognitive impairments in 5×FAD mice. This beneficial effect was associated with a decrease in AD pathologies, including neuronal cell loss and Aβ deposition. Specifically, eugenol inhibited necroptosis activation and increased microglial phagocytosis, which were the underlying mechanisms for the observed reductions in neuronal cell loss and Aβ deposition, respectively. Overall, our data suggest that eugenol would be a potential therapeutic candidate for AD.


Superior sagittal sinus thrombosis with intracerebral hematoma.
Intracranial hematoma secondary to superior sagittal sinus thrombosis is a well described, but rare event. We present a case of interval resolution of a intracerebral hematoma in a patient with thrombosis of the sagittal sinus.


Clinical performance of removable dental prostheses in the moderately reduced dentition: a systematic literature review.
Evidence-based therapeutic recommendations for removable dental prostheses are still lacking. The aim of the present study was a systematic review and meta-analysis of the survival rates of removable dentures in the moderately reduced dentition. In 2014, a systematic literature search in established medical databases (MEDLINE, EMBASE, BIOSIS, SciSearch, Cochrane, FIZ Technik Web) and a hand search of relevant dental journals were conducted. The search terms were relevant MeSH terms, free search terms, and combinations of the two. The search included RCTs, prospective and retrospective studies on survival rates of removable dental prostheses in the moderately reduced dentition with at least 15 participants, an observation period of at least 2 years, and a dropout rate of less than 25 %. The selection of relevant publications was carried out at the title, abstract, and full-text level by at least two of the authors involved. The publications included were tabulated and analyzed. Of the original 12,994 matches, 1923 were analyzed by title, 650 by abstract, and 111 according to the full text. The final review included 19 publications, of which 6 were multiple publications. Cast-metal framework dentures exhibited failure rates of between 33 and 50 % after 5 years. One study with a 25-year observation period reported failure rates of 50 %. Better results were obtained with proper pretreatment and a good recall scheme. Bilateral attachment prostheses showed failure rates of between 11 and 30 % after 5 years. Unilateral attachment prostheses showed failure rates of 75 % after 5 years. Double-crown prostheses dentures show failure rates of 0 to 21.7 % after 3 to 6 years. Heterogeneous study designs and data analyses rendered a meta-analysis impossible, so that an evaluation at the highest level of evidence could not be performed. Within the limitations of this study, it would be correct to state that removable dental prostheses, given suitable pretreatment and follow-up regimes, can provide satisfactory solutions. Based on only one paper, they revealed acceptable results even over a very long observation period (25 years).


Transplantation of VEGFl65-overexpressing vascular endothelial progenitor cells relieves endothelial injury after deep vein thrombectomy.
The purpose of this study was to explore the therapeutic efficacy of VEGF165-overexpressing vascular endothelial progenitor cells (EPCs) in post-thrombotic syndrome. A thrombus model was developed to mimic the in-vivo setting, and adenovirus transfection was used to overexpress VEGF165 in EPCs. These cells were transplanted into the animal model, and their ability to relieve endothelial injury was evaluated using haematoxylin and eosin staining, immunohistochemistry and scanning electron microscopy. Ferric chloride was used to build rat models of the inferior vena cava thrombosis, and HEK 293A cells were used to amplify adenovirus that overexpresses VEGF165. EPCs were infected with adenovirus, and this was confirmed by fluorescence microscopy. Transplantation of VEGH165-overexpressing EPCs into injured endothelial sites led to faster repair of the post-thrombotic tunica intima than wild-type EPCs. Transplantation of VEGF165-overexpressing EPCs was found to promote repair of the tunica intima, thus improving rehabilitation after surgery.


Elective tracheostomy for a patient with a history of difficult intubation.
The anesthetic challenge of managing a difficult airway is demanding under the best of conditions. An emergency operation compounds the difficulty. Seldom do we have the opportunity to truly plan for a possible emergency. This report presents a solution to the problem of knowing of a difficult airway but having no control over the timing or possible emergent state of the operation.


Pathogenicity and immunogenicity of gI/gE/TK-gene-deleted Felid herpesvirus 1 variants in cats.
Felid herpesvirus 1 (FHV-1) is a major pathogenic agent of upper respiratory tract infections and eye damage in felines worldwide. Current FHV-1 vaccines offer limited protection of short duration, and therefore, do not reduce the development of clinical signs or the latency of FHV-1. To address these shortcomings, we constructed FHV ∆gIgE-eGFP, FHV ∆TK mCherry, and FHV ∆gIgE/TK eGFP-mCherry deletion mutants (ΔgI/gE, ΔTK, and ΔgIgE/TK, respectively) using the clustered regularly interspaced palindromic repeats (CRISPR)/CRISP-associated protein 9 (Cas9) system (CRISPR/Cas9), which showed safety and immunogenicity in vitro. We evaluated the safety and efficacy of the deletion mutants administered with intranasal (IN) and IN + subcutaneous (SC) vaccination protocols. Cats in the vaccination group were vaccinated twice at a 4-week interval, and all cats were challenged with infection 3 weeks after the last vaccination. The cats were assessed for clinical signs, nasal shedding, and virus-neutralizing antibodies (VN), and with postmortem histological testing. Vaccination with the gI/gE-deleted and gI/gE/TK-deleted mutants was safe and resulted in significantly lower clinical disease scores, fewer pathological changes, and less nasal virus shedding after infection. All three mutants induced virus-neutralizing antibodies after immunization. In conclusion, this study demonstrates the advantages of FHV-1 deletion mutants in preventing FHV-1 infection in cats.


Personality dependent effects of the ACTH 4--10 fragment on test performances and on concomitant autonomic reactions.
The effect of the ACTH-fragment 4--10 (30 mg SC) on a mental performance test and on some concomitant cardiovascular changes was investigated in comparison with a placebo in a double blind cross-over study. The subjects were either mainly extraverted or mainly introverted according to Eysenck's 'Maudsley Personality Inventory'. Under the influence of the heptapeptide extraverted subjects achieved a higher total score in the mental task performance with a smaller increase of forearm blood flow and of heart rate than under the influence of the placebo. In contrast, under the influence of the placebo introverted subjects achieved a higher total score in the mental task performance with a smaller increase of forearm blood flow and of heart rate than under the influence of the ACTH-fragment. Personality, therefore, determines to some degree how this centrally acting heptapeptide influences efficiency in the mental task performance and the concomitant cardiovascular changes.


Thermal diffuse scattering in sub-angstrom quantitative electron microscopy-phenomenon, effects and approaches.
This paper reviews the recent progress in the following areas. (1) In quantitative high-resolution transmission electron microscopy, the theoretically calculated images usually give better contrast than the experimentally observed ones although all of the factors have been accounted for. This discrepancy is suggested due to thermal diffusely scattered (TDS) electrons, which were not included in the image calculation. The contribution from TDS electrons is especially important if the image resolution is approaching 0.1 nm and beyond with the introduction of Cs corrected microscopes. A more rigorous multislice theory has been developed to account for this effect. (2) We proved that the off-axis holography is an ideal energy filter that even filters away the contribution made by TDS electrons in the electron wave function, but conventional high-resolution microscopy do contain the contribution made by phonon scattered electrons. (3) In electron scattering, most of the existing dynamical theories have been developed under the first order diffuse scattering approximation, thus, they are restricted to cases where the lattice distortion is small. A formal dynamical theory is presented for calculating diffuse scattering with the inclusion of multiple diffuse scattering. By inclusion of a complex potential in dynamical calculation, a rigorous proof is given to show that the high order diffuse scattering are fully recovered in the calculations using the equation derived under the distorted wave Born approximation, and more importantly, the statistical time and structure averages over the distorted crystal lattices are evaluated analytically prior numerical calculation. This conclusion establishes the basis for expanding the applications of the existing theories. (4) The 'frozen lattice' model is a semi-classical approach for calculating electron diffuse scattering in crystals arisen from thermal vibration of crystal atoms. Based on a rigorous quantum mechanical phonon excitation theory, we have proved that the frozen lattice mode is an excellent approximation and no detectable error would be possible under normal experimental conditions.


Translocation t(5;14)(q35;q32) in three cases of childhood T cell acute lymphoblastic leukemia: a new recurring and cryptic abnormality.
We report three cases of T-ALL in which conventional cytogenetic analysis yielded normal karyotypes, but for which a new M-FISH technique (IPM-FISH) was able to detect a translocation. For these patients this technique highlighted a new, recurring and cryptic translocation t(5;14)(q35;q32) in childhood T-ALL which might be phenotypically restricted. The most innovative part of this technique is the use of interspersed polymerase chain reaction (IRS-PCR) painting probes that show an R-band pattern simultaneous with the combinatorial labeling. Contrary to the DOP-PCR, IRS-PCR-derived probes provide stronger hybridization signals at the telomeric ends that potentially increase the possibility of detecting cryptic translocations. All the IPM-FISH findings were validated by FISH with whole chromosome painting and unique sequence probes. These results demonstrate the efficient use of IPM-FISH as an improved, single-step method for the identification of cryptic chromosomal abnormalities. This new IPM-FISH technique is a good tool to display cryptic chromosomal abnormalities.


Role of lipids in the interaction of antimicrobial peptides with membranes.
Antimicrobial peptides (AMPs) take part in the immune system by mounting a first line of defense against pathogens. Recurrent structural and functional aspects are observed among peptides from different sources, particularly the net cationicity and amphipathicity. However, the membrane seems to be the key determinant of their action, either as the main target of the peptide action or by forming a barrier that must be crossed by peptides to target core metabolic pathways. More importantly, the specificity exhibited by antimicrobial peptides relies on the different lipid composition between pathogen and host cells, likely contributing to their spectrum of activity. Several mechanisms of action have been reported, which may involve membrane permeabilization through the formation of pores, membrane thinning or micellization in a detergent-like way. AMPs may also target intracellular components, such as DNA, enzymes and even organelles. More recently, these peptides have been shown to produce membrane perturbation by formation of specific lipid-peptide domains, lateral phase segregation of zwitterionic from anionic phospholipids and even the formation of non-lamellar lipid phases. To countermeasure their activity, some pathogens were successful in developing effective mechanisms of resistance to decrease their susceptibility to AMPs. The functional and integral knowledge of such interactions and the clarification of the complex interplay between molecular determinants of peptides, the pathogen versus host cells dichotomy and the specific microenvironment in which all these elements convene will contribute to an understanding of some elusive aspects of their action and to rationally design novel therapeutic agents to overcome the current antibiotic resistance issue.


Activity of preoptic neurons during synchronization and desynchronization.
Extracellular unit activity from 29 neurons in the preoptic area was recorded together with the cortical EEG in encéphale isolé cats. A majority (55%) of neurons showed alterations in their firing rates during transient changes in the EEG. Among them, a majority (62.5%) showed an increased firing rate during synchronization and the remaining showed an increased firing rate during desynchronization of the EEG. Most neurons showed a Poisson distribution pattern of firing during both the synchronized and the desynchronized phases of the EEG. The changes in the neuronal discharge occurring together with the specific changes in the cortical EEG fits in well with the assigned role of the preoptic area in the sleep-waking cycle.


International standards for health economic evaluation with a focus on the German approach.
Health economic evaluation (HEE) is increasingly used in healthcare decision-making on the allocation of limited resources in national healthcare systems. Although the methods used for HEE vary in different countries, all economic evaluations address two questions: Are limited resources used optimally? Is value for money achieved in their use? Our objective is to explain some fundamental concepts in HEE and how these concepts are adapted in different countries, notably in Germany. We performed a bibliographic search to identify existing methods of health economic evaluation of new drugs used by the official agencies of 11 countries (Austria, Australia, Canada, Finland, France, the Netherlands, Norway, New Zealand, Sweden, the United States and England and Wales) and compared them with that used by the German national agency IQWiG. All countries considered follow internationally established standards of HEE. The majority of countries, including Germany, utilize primary outcome parameters such as disease-related morbidity and mortality for assessing relative efficacy and effectiveness. The most frequently recommended form of health economic evaluation is the cost-utility analysis (CUA). The German IQWIG is the only HTA body to use the cost-benefit concept of 'efficiency frontier' in its assessment. While the core principles of HEE are the same worldwide, there is a lack of harmonization in the details. This requires resource-consuming adaptations in the analyses to meet different national requirements. We describe the core principles of HEE as a common basis for further discussions by all stakeholders.


The role of spirituality and religiosity on the cognitive decline of community-dwelling older adults: a 4-year longitudinal study.
This study aims to assess whether having religious beliefs, attending religious services and using spiritual-religious coping (SRC) are longitudinally associated with cognitive decline in community-dwelling older adults. A 4-year longitudinal study of 261 Brazilian older adults was conducted. Hierarchical adjusted linear regression models and logistic regression models were performed to evaluate the longitudinal effects of religious beliefs, church attendance and positive and negative SRC on cognitive decline. The findings revealed that spiritual and religious beliefs at baseline could affect cognitive function in two different ways. When older adults use religiosity and spirituality (RS) in a functional positive manner, it results in better cognitive outcomes and a slower rate of decline. However, when used in a negative manner, it results in worse cognitive outcomes. These results could help health care professionals address SRC among older adults, particularly those at a higher risk of cognitive decline. Considering that RS is very important for older adults, health care professionals should be aware of the beliefs of their patients and address RS in clinical practice.


Eosinophil-derived galectin-10 upregulates matrix metalloproteinase expression in bullous pemphigoid blisters.
Bullous pemphigoid (BP) is an autoimmune bullous disease in which abundant eosinophils accumulate in the blisters. Galectin-10 abounds in the cytoplasm of eosinophils and is released as a result of eosinophil extracellular trap cell death (EETosis). To identify EETosis and the pathological roles of galectin-10 in BP. EETosis and galectin-10 in BP blisters were confirmed by immunofluorescence and transmission electron microscopy. The concentrations of galectin-10 in serum and blister fluid from BP patients were studied by ELISA. The matrix metalloproteinase (MMP) expression in BP blisters was immunohistochemically compared to that in healthy controls. As an in vitro assay, normal human epidermal keratinocytes (NHEKs) and normal human dermal fibroblasts (NHDFs) were stimulated with galectin-10, followed by MMP expression measurement by real-time PCR and ELISA. The signaling pathways activated by galectin-10 were studied using Western blotting and confirmed by inhibition assays. Galectin-10-containing eosinophil infiltration and the extracellular deposition of major basic protein were observed in BP blisters. The ultrastructural characteristics of tissue eosinophils indicated piecemeal degranulation and EETosis. In the BP patients, the concentration of galectin-10 was higher in the blister fluid than in the serum. Several types of MMPs were upregulated in BP blisters. Galectin-10 upregulated the production of MMPs through the pathways of p38 MAPK, ERK and JNK in NHEKs and NHDFs. In the BP blisters, the eosinophils underwent EETosis and released galectin-10. Galectin-10 might contribute to BP blister formation through the production of MMPs by keratinocytes and fibroblasts.


Oscillations, clocks and segmentation.
Notch signalling molecules, such as the basic helix-loop-helix factors Hes1 and Hes7, periodically change their expression in the presomitic mesoderm, and each cycle of gene expression is associated with somite formation (every two hours in mouse). This cyclic expression is the manifestation of an intrinsic mechanism, called the segmentation clock, which is essential for coordinated somite segmentation. Interestingly, the oscillatory expression of Hes1 is observed in many cell types after serum stimulation, suggesting that this ultradian clock is not unique to presomitic mesoderm cells but widely distributed. This oscillation depends on the negative feedback loop, and once its promoter is constitutively activated, Hes1 seems to start oscillatory gene expression autonomously. Thus, Hes1 acts as a device that transduces a direct current of input into an alternating current, which ticks the hours in many biological systems.


Investigating the initial steps of auricin biosynthesis using synthetic biology.
Streptomyces lavendulae subsp. lavendulae CCM 3239 (formerly Streptomyces aureofaciens CCM 3239) contains a type II polyketide synthase (PKS) biosynthetic gene cluster (BGC) aur1 whose genes were highly similar to angucycline BGCs. However, its product auricin is structurally different from all known angucyclines. It contains a spiroketal pyranonaphthoquinone aglycone similar to griseusins and is modified with D-forosamine. Here, we describe the characterization of the initial steps in auricin biosynthesis using a synthetic-biology-based approach. We have created a plasmid system based on the strong kasOp* promoter, RBS and phage PhiBT1-based integration vector, where each gene in the artificial operon can be easily replaced by another gene using unique restriction sites surrounding each gene in the operon. The system was validated with the initial landomycin biosynthetic genes lanABCFDLE, leading to the production of rabelomycin after its integration into Streptomyces coelicolor M1146. However, the aur1DEFCGHA homologous genes from the auricin aur1 BGC failed to produce rabelomycin in this system. The cause of this failure was inactive aur1DE genes encoding ketosynthases α and β (KSα, KSβ). Their replacement with homologous aur2AB genes from the adjacent aur2 BGC resulted in rabelomycin production that was even higher after the insertion of two genes from the aur1 BGC, aur1L encoding 4-phosphopantetheinyl transferase (PPTase) and aur1M encoding malonyl-CoA:ACP transacylase (MCAT), suggesting that Aur1L PPTase is essential for the activation of the acyl carrier protein Aur1F. These results suggest an interesting communication of two BGCs, aur1 and aur2, in the biosynthesis of the initial structure of auricin aglycone.


New Consideration of Herlyn-Werner-Wunderlich Syndrome Diagnosed by Ultrasound.
As a rare malformation of the female reproductive system, Herlyn-Werner-Wunderlich syndrome (HWWS) was categorized into 3 classifications. It was recommended recently that, on the basis of the past classification, cervicovaginal atresia without communicating uteri should be newly added as classification 4. The surgical intervention will differ by type. To better optimize patient counseling as well as the preoperative evaluation and planning, our objective was to describe the ultrasound characteristics of each type of HWWS, including the new type. From January 1995 to November 2015, 37 cases of HWWS in with complete ultrasound information confirmed by surgery in the Peking Union Medical College Hospital were reviewed. We analyzed their ultrasound features, including hematometra, hematocervix, hematocolpos, and an ovarian chocolate cyst. All of the ultrasound images of the 37 patients showed uterus didelphys with ipsilateral renal agenesis. Compared with the other 3 types, classification 4 showed distinctive ultrasound characteristics. Most cases of classification 4 showed hematometra (5 of 7 [71.4%]) and an ipsilateral ovarian chocolate cyst (6 of 7 [85.7%]), which was significantly higher than in the other 3. A rudimentary uterine horn was also a distinctive characteristic in this type. Meanwhile none of the classification 4 cases showed hematocervix or hydrocolpos, which were common signs of the other 3. According to this new classification criteria for HWWS, ultrasound characteristics of the new classification 4 differ from the others. As classification 4 was suggested to have a different surgical option, we should pay attention to its ultrasound characteristics, which might help in providing more information about the treatment and prognosis to the gynecologist.


Lifelong Tone Language Experience does not Eliminate Deficits in Neural Encoding of Pitch in Autism Spectrum Disorder.
Atypical pitch processing is a feature of Autism Spectrum Disorder (ASD), which affects non-tone language speakers' communication. Lifelong auditory experience has been demonstrated to modify genetically-predisposed risks for pitch processing. We examined individuals with ASD to test the hypothesis that lifelong auditory experience in tone language may eliminate impaired pitch processing in ASD. We examined children's and adults' Frequency-following Response (FFR), a neurophysiological component indexing early neural sensory encoding of pitch. Univariate and machine-learning-based analytics suggest less robust pitch encoding and diminished pitch distinctions in the FFR from individuals with ASD. Contrary to our hypothesis, results point to a linguistic pitch encoding impairment associated with ASD that may not be eliminated even by lifelong sensory experience.


Calcineurin inhibitors and post-transplant weight gain.
Excessive weight gain that leads to obesity is quite common after kidney transplantation. This is often attributed to immunosuppression. The aim of this retrospective study was to assess the effect of calcineurin inhibitors on post-transplant weight gain. A total of 99 patients were studied. The patients were divided into cyclosporine A (CyA) and tacrolimus (Tac) groups and were evaluated for weight changes and risk factors related to weight gain. The weights of patients in both groups significantly increased after the sixth month. The median weight gain at 12 months was 3.5 and 8.0 kg compared with pretransplant dry weight in the Tac and CyA groups, respectively. The increases in the CyA group were significant compared with those of the Tac group. The prevalences of obese and overweight patients in both groups did not differ during a 12-month follow-up. The frequencies of diabetes mellitus, hypertension and dyslipidemia were comparable in both groups. The decrease in systolic blood pressure (BP) of the Tac group was significant compared with the decrease in the CyA group at the 12th month. In the 12-month follow-up period, the increases in triglyceride, total- and low-density lipoprotein-cholesterol values of the CyA group were significantly higher than those of the Tac group. The weight change between 0 and 12 months was negatively correlated with pretransplant body mass index (BMI) and positively with cumulative corticosteroid doses, total-cholesterol and BP changes. Only pretransplant BMI, creatinine clearance, CyA usage, being hypertensive and dyslipidemic were independent predictors of weight gain at the 12th month. Our results suggested that the type of immunosuppression may affect post-transplant weight gain.


Aromatase inhibition by R 76713: experimental and clinical pharmacology.
R 76713 is a new non-steroidal compound which inhibits aromatase in vitro and in vivo with a potency of at least 1000-fold that of aminoglutethimide. In male cynomolgus monkeys peripheral conversion of labeled androstenedione to estrone is decreased by 85%, 4-5 h after a single intravenous dose of 0.003 mg/kg of R 76713, without altering steroid metabolic clearance rates. In rats fed a sodium-depleted diet for 3 weeks, plasma levels of aldosterone and plasma renin activity remain unchanged 2 h after a single oral dose of up to 20 mg/kg of R 76713. This confirms previous data on the selectivity of R 76713 for aromatase inhibition as compared to inhibition of other enzymes involved in steroid biosynthesis. In male volunteers, a single oral dose of 5 or 10 mg of R 76713 lowers median plasma estradiol levels from 70 pM to the detection limit of the assay (30 pM) 4 and 8 h after intake, whereas no important changes are detected after placebo administration. In 15 premenopausal female volunteers receiving a single oral dose of 20 mg of R 76713, mean plasma estradiol levels decrease from 415 pM (before) to 179, 149 and 185 pM respectively 4, 8 and 24 h after intake whereas they remain above 380 pM after placebo (n = 7).


Trends in mortality due to diabetes in Brazil, 1996-2011.
Over recent decades, Brazilian mortality registration has undergone increasing improvement in terms of completeness and quality in cause of death reporting. These improvements, however, complicate the description of mortality trends over this period. We aim to characterize the trend in diabetes mortality in Brazil and its five regions in adults (30-69 years), from 1996 to 2011 after corrections for underreporting of deaths and redistribution of ill-defined causes and "garbage codes". Starting with official data from the Brazilian Mortality Information System (SIM) for adults aged 30-69 in the period 1996 to 2011 for diabetes (ICD-10 codes E10-14), we redistributed garbage codes using methods based on the Global Burden of Disease Study (2010), redistributed ill-defined causes based on recent Brazilian investigations of similar cases and corrected for underreporting using official estimates of deaths. With these corrections, age-standardized mortality fell approximately 1.1 %/year for men and 2.2 %/year for women from 1996 to 2011. The rate of decline first accelerated and then decelerated, reaching stable rates in men and minimal declines in women from 2005 onward. Regional inequalities decreased during the period in both relative and absolute terms. Mortality due to diabetes declined in Brazil from 1996 to 2011, minimally in men and considerably in women. The lesser declines in recent years may reflect the increasing prevalence of diabetes, and suggest that current efforts to prevent diabetes and minimize the impact of its complications need to be reinforced to ensure that declines will continue.


Pertussis toxin potentiates anesthesia-induced renin secretion.
The plasma renin concentration was measured after anesthesia in control and in pertussis toxin-treated rats. The anesthetics increased renin secretion and this effect was markedly magnified in rats treated with pertussis toxin. Propranolol partially blocked the increase in plasma renin concentration produced by anesthetics. It is concluded that the adrenergic system is involved in this effect and that pertussis toxin magnifies it by potentiating the beta-adrenergic action.


The interaction effect of sleep deprivation and cannabinoid type 1 receptor in the CA1 hippocampal region on passive avoidance memory, depressive-like behavior and locomotor activity in rats.
Increasing evidence shows the interaction effect of cannabinoids and sleep on cognitive functions. In the present study, we aimed to investigate the interaction effect of cannabinoids type 1 receptor (CB1r) in the CA1 hippocampal region and sleep deprivation (SD) on passive avoidance memory and depressive-like behavior in male Wistar rats. We used water box apparatus to induce total SD (TSD) for 24 h. The shuttle-box was applied to assess passive avoidance memory and locomotion apparatus was applied to assess locomotor activity. Forced swim test (FST) was used to evaluate rat's behavior. ACPA (CB1r agonist) at the doses of 0.01, 0.001 and 0.0001 μg/rat, and AM251 (CB1r antagonist) at the doses of 100, 10 and 1 ng/rat were injected intra-CA1, five minutes after training via stereotaxic surgery. Results showed SD impaired memory. ACPA at the doses of 0.01 and 0.001 μg/rat impaired memory and at all doses did not alter the effect of SD on memory. AM251 by itself did not alter memory, while at lowest dose (1 ng/rat) restored SD-induced memory deficit. Both drugs induced depressive-like behavior in a dose-dependent manner. Furthermore, both drugs decreased swimming at some doses (ACPA at 0.0001 μg/rat, AM251 at 0.001 and 0.01 ng/rat). Also, ACPA at the highest dose increased climbing of SD rats. In conclusion, we suggest CB1r may interact with the effect of SD on memory. Additionally, cannabinoids may show a dose-dependent manner in modulating mood and behavior. Interestingly, CB1r agonists and antagonists may exhibit a similar effect in some behavioral assessments.


catena-Poly[silver(I)-bis-[μ-4-methyl-1H-1,2,4-triazole-3(4H)-thione-κ(2) S:S]-silver(I)-di-μ-thio-cyanato-κ(2) S:N;κ(2) N:S].
In the title one-dimensional coordination polymer, [Ag2(NCS)2(C3H5N3S)2] n , the Ag(I) atom adopts a distorted tetra-hedral AgNS3 geometry. Adjacent Ag(I) atoms in the [001] chain are alternately linked by pairs of bridging 4-methyl-1H-1,2,4-triazole-3(4H)-thione (Hmptrz) ligands (via their S atoms) and double thio-cyanate bridges linking through both S and N atoms (μ-1,3-SCN). An intra-chain N-H⋯N hydrogen bond occurs between the NH group of the triazole ring and the N atom of the thio-cyanate bridging ligand. A (101) sheet structure arises from inter-chain S⋯N short contacts [3.239 (3) Å] involving the thio-cyanate S atom and the triazole-ring N atom and possible very weak π-π stacking [centroid-centroid separation = 4.0762 (18) Å] between the triazole rings.


Different reactivity of activated human B cells to B-cell growth factor and interleukin 2 in the costimulation assay with anti-IgM antibody and in the preactivation assay with Staphylococcus bacteria.
The two main assay systems which have been developed for the study of lymphokine-mediated human B-cell proliferation, i.e., the costimulation assay with anti-mu antibody and the preactivation assay with Staphylococcus aureus Cowan I (SAC) bacteria, were compared. Purified interleukin 2 (IL-2), obtained by the recombinant DNA technology (r-IL-2), enhanced the proliferative response of anti-mu-stimulated human B cells in the costimulation assay with anti-mu antibody and maintained the B-cell proliferation induced by preactivation with SAC bacteria. Although the majority of T-cell clones, established from normal peripheral blood T lymphocytes, showed production of both IL-2 and B-cell growth factor (BCGF) following phytohemagglutinin (PHA)-stimulation, some T-cell clones were found whose supernatants (PHA-SN), apparently free of IL-2, manifested strong BCGF activity in the costimulation assay with anti-mu antibody. However, the same clonal, IL-2-free, T-cell SN displayed no BCGF activity in the preactivation assay with SAC bacteria. When B cells were activated for 3 days with anti-mu antibody, followed by the addition of r-IL-2 or clonal T-cell SN containing BCGF for an additional 3 days, r-IL-2 showed the ability to maintain B-cell proliferation, whereas clonal SN containing BCGF had virtually no effect. These data indicate that the costimulation assay with anti-mu antibody explores the reactivity of normal human B cells to both BCGF and IL-2, whereas the preactivation assay with SAC bacteria, due to a shorter reactivity to BCGF of activated human B cells, essentially represents a probe for the study of IL-2-promoted B-cell proliferation.


Moose herbivory, browse quality, and nutrient cycling in an Alaskan treeline community.
Moose (Alces alces) browsing on diamondleaf willow (Salix planifolia pulchra) caused significant increases in subsequent growth of stems and leaves in treeline plant communities in central Alaska, USA. Willows growing in the shade were significantly more palatable for moose than those growing in the sun. Moose density had strong effects on rates of nutrient cycling, ostensibly through effects of browsing and inputs from fecal and urinary nitrogen. Moose are a keystone herbivore that likely mediate rates of nutrient cycling in northern ecosystems.


Child sexual abuse knowledge/attitude scale for parents: Reliability and validity of the Turkish version.
The aim of this study was to determine the validity and reliability of the Turkish version of the child sexual abuse knowledge/attitude scale for parents (CSAKAS). This methodological study was carried out with 193 parents in between February and May 2018. The validity and reliability of the six-factors structure scale were confirmed. The Cronbach's alpha coefficient of the whole scale is 0.793, the item-total score correlations ranged between 0.341 and 0.719 and the test-retest coefficient was 0.841. The CSAKAS Turkish scale was found to be a valid and reliable tool that can be used to evaluate the knowledge and attitudes of parents about child sexual abuse.


The aminoglycosides: streptomycin, kanamycin, gentamicin, tobramycin, amikacin, netilmicin, and sisomicin.
Despite the introduction of newer, less toxic antimicrobial agents, the aminoglycosides remain useful in the treatment of serious, hospital-acquired, gram-negative bacillary infections, especially those caused by Pseudomonas aeruginosa. Formidable nephrotoxicity and ototoxicity have limited the use of neomycin to topical or oral administration. Widespread antimicrobial resistance among Enterobacteriaceae has restricted the use of streptomycin and kanamycin to a few specific clinical situations. Gentamicin, tobramycin, and amikacin are active against a wide range of Enterobacteriaceae and many P. aeruginosa organisms. In medical centers where gentamicin resistance is prevalent, amikacin is the aminoglycoside of choice. Fortunately, amikacin resistance has not seemed to increase substantially, even in institutions where usage has been extensive for a long period. No new aminoglycoside has proved to be superior to amikacin.


Identification of the perfect state of Cryptococcus neoformans from 195 clinical isolates including 84 from AIDS patients.
Filobasidiella neoformans is the teleomorphic state of Cryptococcus neoformans and it is a heterothalic. The purpose of this study was to establish the proportions of each mating types (a, alpha) from among 195 strains of C. neoformans isolated from clinical material. The culture medium used was sunflower agar. Cultures were incubated at 20-22 degrees C for 15 days and observed periodically for one month. Non-reactive strains were mated several times with different reactive strains. Under these conditions 96.8% of the strains were found to be reactors. Among both varieties of C. neoformans, mating type alpha was found to have the highest frequency of 95% in the variety neoformans and 84% in the variety gattii. These results showed a higher reactivity in comparison with other investigators. This difference could be due to the medium used or to repeated mating with different reactive tested strains.


Cellular mechanisms by which oxytocin mediates uterine prostaglandin F2 alpha synthesis in bovine endometrium: role of calcium.
The objective of these experiments was to determine the role of Ca2+ during oxytocin-stimulated prostaglandin (PG) F2 alpha release from bovine endometrial tissue in vitro. Uteri were collected from dairy cows on the day after spontaneous luteal regression. Caruncular endometrial explants were dissected and incubated in vitro to determine phospholipase C activity or PGF2 alpha release. A23,187 (a calcium ionophore) and maitotoxin (an activator of voltage-gated L-type calcium channels) stimulated release of PGF 2 alpha in a concentration-dependent manner (P < 0.05). Thapsigargin (induces accumulation of Ca2+ in the cytoplasm by inhibiting endoplasmic reticulum Ca2+/ATPase pumps) stimulated release of PGF2 alpha in a concentration-dependent manner as well (P < 0.13). Oxytocin (10(-6) M), AIF4- (a nonspecific activator of G-proteins; 10(-5) M), A23,187 (10(-5) M), and melittin (a stimulator of phospholipase A2; 10(-4) M) stimulated PGF2 alpha release when explants were incubated in Ca(2+)-free medium (P < 0.10); however, oxytocin, A23,187, or melittin were unable to stimulate PGF2 alpha release when explants were incubated in Ca(2+)-free medium containing the calcium chelator EGTA (P < 0.10). This treatment did not prevent oxytocin or AIF4- from stimulating phospholipase C activity (P < 0.08). CoCl2 (a nonspecific Ca2+ channel blocker) and methoxyverapamil (a specific voltage-gated L-type Ca2+ channel blocker) prevented oxytocin from stimulating PGF2 alpha release (P < 0.05). Our results suggest that both extracellular and intracellular Ca2+ may be required for oxytocin to stimulate PGF2 alpha secretion in bovine endometrial tissue.


Prefrontal cortex-specific Dcc deletion induces schizophrenia-related behavioral phenotypes and fail to be rescued by olanzapine treatment.
Multiple genome studies have discovered that variation in deleted in colorectal carcinoma (Dcc) at transcription and translation level were associated with the occurrences of psychiatric disorders. Yet, little is known about the function of Dcc in schizophrenia (SCZ)-related behavioral abnormalities and the efficacy of antipsychotic drugs in vivo. Here, we used an animal model of prefrontal cortex-specific knockdown (KD) of Dcc in adult C57BL/6 mice to study the attention deficits and impaired locomotor activity. Our results supported a critical role of Dcc deletion in SCZ-related behaviors. Notably, olanzapine rescued the SCZ-related behaviors in the MK801-treated mice but not in the cortex-specific Dcc KD mice, indicating that Dcc play a critical in the mechanism of antipsychotic effects of olanzapine. Knockdown of Dcc in prefrontal cortex results in glutamatergic dysfunction, including defects in glutamine synthetase and postsynaptic maturation. As one of the major risk factors of the degree of antipsychotic response, Dcc deletion-induced glutamatergic dysfunction may be involved in the underlying mechanism of treatment resistance of olanzapine. Our findings identified Dcc deletion-mediated SCZ-related behavioral defects, which serve as a valuable animal model for study of SCZ and amenable to targeted investigations in mechanistic hypotheses of the mechanism underlying glutamatergic dysfunction-induced antipsychotic treatment resistance.


Preparation and characterization of an improved Cu(2+)-cyclen polyurethane material that catalyzes generation of nitric oxide from S-nitrosothiols.
A new, stable and highly efficient Cu(2+)-cyclen-polyurethane material is described and shown to exhibit improved performance compared to prior materials for the catalytic decomposition of S-nitrosothiols to physiologically active nitric oxide.


Renal failure and hyponatremia in patients with cirrhosis and skin and soft tissue infection. A retrospective study.
Skin and soft tissue infection in cirrhosis is considered a non-severe infection, but specific information is lacking. This study aimed at assessing the characteristics, occurrence of renal failure, and outcome of cirrhotic patients with skin and soft tissue infection. Ninety-two patients with cirrhosis and skin and soft tissue infection admitted to hospital within a 6-year period were retrospectively analyzed. A control group matched by severity of liver disease, admitted for reasons other than infection, was also studied. Resolution of the infection was achieved in 96% of patients. Twenty (21.7%) patients with skin and soft tissue infection developed renal failure, compared to only five patients (5.4%) of the control group (p=0.001). Renal failure was persistent despite infection resolution in 10 of the 20 patients vs. none of the control group. Renal failure was associated with poor prognosis. Hyponatremia developed in 40% and 25% of the infection and control group, respectively (p=0.028). Within a 3-month follow-up period, 25 patients (23%) with skin and soft tissue infection died or were transplanted compared to only four patients (4%) of the control group (p<0.001). Factors independently associated with mortality in the infection group were: site of acquisition of the infection and MELD-sodium score at diagnosis. Skin and soft tissue infection is a severe complication of cirrhosis with high frequency of renal failure and hyponatremia that may persist despite resolution of the infection. MELD-sodium score is useful to assess 3-month mortality in these patients.


Time-resolved diffusion reveals photoreactions of BLUF proteins with similar functional domains.
BLUF (blue light sensor using flavin) proteins are the blue light receptors that consist of flavin-binding BLUF domains and functional domains. Upon blue light excitation, the hydrogen bond network around the flavin chromophore changes, and the absorption spectrum in the visible region shifts to red. Light signal received in the BLUF domain is intramolecularly or intermolecularly transmitted to the functional region. In this review, the reactions of three BLUF proteins with similar EAL functional groups within the protein (BlrP1, and YcgF), or with a separated target protein (PapB) are described using time-resolved diffusion technique. The diffusion coefficients (D) of the BLUF domains did not significantly change upon photoexcitation, whereas those of the full-length proteins BlrP1 and YcgF and the PapB-PapA system significantly decreased. The changes in D should be due to diffusion-sensitive conformational changes (DSCC) that alter the friction of diffusion. The time constants of the major D changes of BlrP1 and PapB-PapA were similar (~ 20 ms), although the magnitude of the friction change depended on the proteins. Similarities and differences among the reactions of these proteins were clarified from the viewpoint of DSCC.


Primordial Prevention of Atherosclerotic Cardiovascular Disease: A REVIEW OF THE LITERATURE.
The primordial prevention of atherosclerotic cardiovascular disease (ASCVD) involves the prevention of the onset of its risk factors. This review explores the associations between early modifiable risk factors and the development of ASCVD in adulthood, as well as evidence-based interventions to prevent them. A review was conducted on the basis of an in-depth literature search including longitudinal observational data, systematic reviews and meta-analyses published in 2012 or later, clinical trials, and additional manual searches of recent literature based on reference lists of other reviews and relevant guidelines. ASCVD is a disease that begins in childhood; hence, primordial prevention is an important target for improving cardiovascular morbidity and mortality later in life. Data from large-scale population studies have consistently identified the following modifiable risk factors for the development of ASCVD: smoking, overweight and obesity, high cholesterol, high blood pressure, hyperglycemia, poor diet, and physical inactivity. These risk factors originate during the prenatal, childhood, and adolescent stages of life. Various successful interventions to prevent the onset of each risk factor have been evaluated at the individual, community, and population levels. Implementation of a heart-healthy dietary pattern and regular exercise early in life are large components of many successful interventions.


A new approach for antimicrobial and antiviral activities of biocompatible nanocomposite based on cellulose, amino acid and graphene oxide.
In this work, biocompatible, antimicrobial, and antiviral nanocomposites were prepared through two steps. In the first step, periodate oxidation of cellulose was performed to get dialdehyde cellulose (DAC). The second step included the reaction of DAC with sulfur-containing amino acids included Cysteine (Cys) and Methionine (Meth) in the presence of graphene oxide (GO). The prepared nanocomposites were characterized via FT-IR, SEM, TEM, and TGA. Antimicrobial and antiviral activities for all designed nanocomposites besides DAC were carried out. Both DAC/GO/Cys and DAC/GO/Meth exhibited a promising antimicrobial activity against Gram-negative (E. coli and P. aeruginosa), Gram-positive (B. subtilis and S. aureus), and unicellular fungi (C. Albicans and C. neoformans), while the DAC/GO/Cys/Meth nanocomposite was the lowest. Moreover, all designed nanocomposites have a strong antiviral activity against Herpes simplex virus 1(HSV-1) at minimum nontoxic concentration. Additionally, Computational procedures and Molecular docking showed the reactivity and stability of the molecules that have biological activity against Gram-positive, Gram-negative, and HSV-1. As well as DAC incorporation with amino acid enhanced their reactivity and their interaction.


Close association of endothelial dysfunction with insulin resistance and carotid wall thickening in hypertension.
Endothelial dysfunction has been regarded as an early stage in the atherosclerotic process. Endothelial dysfunction and insulin resistance were observed in hypertensive subjects and were associated with carotid wall thickening. We examined the determinants of endothelial dysfunction including insulin sensitivity and carotid wall thickening. A total of 41 subjects with nondiabetic essential hypertension were studied. Endothelial function of brachial artery and carotid wall thickening were assessed noninvasively using ultrasound technique. In brachial artery, we measured flow-mediated endothelium-dependent vasodilation (FMD) and glyceryl trinitrate-induced endothelium-independent vasodilation (GTN). We estimated intima-media thickness of the common carotid artery (IMT). Insulin sensitivity was measured according to the steady-state plasma glucose (SSPG) method. High SSPG levels indicated insulin resistance. On univariate analysis, there were significant negative correlations between FMD and SSPG (r = -0.695, P <.0001) or IMT (r = -0.449, P <.004). The FMD was negatively correlated significantly with age and with systolic and diastolic blood pressures (BP). A significant negative correlation was observed between GTN and SSPG. There was a significant positive relation between SSPG and IMT. On multiple regression analysis including systolic BP, SSPG, and age as independent variables and FMD as a dependent variable, FMD was independently related to SSPG (P <.03) and systolic BP (P <.02). If the presence of SSPG, diastolic BP, and age were entered as independent variables against FMD, FMD was independently related to SSPG (P <.002). One of the major determinants of endothelial function was insulin resistance. Our findings suggest that endothelial dysfunction and early structural vascular changes were related to insulin resistance.


Hyperbaric Oxygen: an important treatment modality in severe hemorrhagic cystitis after allogeneic hematopoietic stem cell transplantation.
Hemorrhagic cystitis (HC) is a generally self-limited complication of hematopoietic stem cell transplantation (HSCT). It may occur in the early or late posttransplant period and can promote sometimes severe morbidity. We analyzed our data regarding HC in allogeneic HSCT patients in order to establish the efficacy of hyperbaric oxygen (HBO) therapy in severe HC and to document the main problems during its use. Between March 1993 and August 2006, 161 patients received allogeneic HSCT. Mesna, hyperhydration and forced diuresis were used as early HC prophylaxis of cyclophosphamide-induced HC. However, HC was diagnosed in 49 of the 161 recipients and 17 of them were considered as severe HC. We analyzed their data retrospectively. Forced diuresis with hyperhydration (up to 8 L/day) and transfusion support to maintain a platelet count above 30x109/L were sufficient in 10 of the 17 patients with severe HC. Alternative therapies used included intravesical irrigation with formalin and prostaglandin (PG)F2 alpha and HBO, and HBO appeared to be the most useful among them. We conclude that HBO offers a noninvasive therapeutic alternative in the management of intractable HC in the HSCT setting.