Molecular epidemiology of Streptococcus agalactiae isolated from mastitis in Brazilian dairy herds.
Streptococcus agalactiae is one of the most common pathogens leading to mastitis in dairy herds worldwide; consequently, the pathogen causes major economic losses for affected farmers. In this study, multilocus sequence typing (MLST), genotypic capsular typing by multiplex polymerase chain reaction (PCR), and virulence gene detection were performed to address the molecular epidemiology of 59 bovine (mastitis) S. agalactiae isolates from 36 dairy farms located in the largest milk-producing mesoregions in Brazil (Minas Gerais, São Paulo, Paraná, and Pernambuco). We screened for the virulence genes bac, bca, bibA, cfb, hylB, fbsA, fbsB, PI-1, PI-2a, and PI-2b, which are associated with adhesion, invasion, tissue damage, and/or immune evasion. Furthermore, five capsular types were identified (Ia, Ib, II, III, and IV), and a few isolates were classified as non-typeable (NT). MLST revealed the following eight sequence types (STs): ST-61, ST-67, ST-103, ST-146, ST-226, ST-314, and ST-570, which were clustered in five clonal complexes (CC64, CC67, CC103, CC17, and CC314), and one singleton, ST-91. Among the virulence genes screened in this study, PI-2b, fbsB, cfb, and hylB appear to be the most important during mastitis development in cattle. Collectively, these results establish the molecular epidemiology of S. agalactiae isolated from cows in Brazilian herds. We believe that the data presented here provide a foundation for future research aimed at developing and implementing new preventative and treatment options for mastitis caused by S. agalactiae.


[Postchemotherapy surgery in patients with non-seminomatous testicular tumors].
From 1982 to 1993, 220 patients with metastatic nonseminomatous germ cell testicular tumours were treated with a combination of cisplatin, vinblastine and bleomycin. Complete remission was achieved in 108 (49.1%) pts. on chemotherapy alone. Residual abdominal or pulmonary masses persisted on completion of chemotherapy in 85 (38.6%) pts. All of them had normal serum levels of tumour markers. In 13 (5.9%) pts tumour markers were positive. Fourteen (6.4%) pts died during primary chemotherapy. The pts with residual masses and normal tumour markers were treated by surgical exploration and masses removal. In 62 pts the masses were resected through laparotomy, in 19 pts through pulmonary surgery and in four pts laparotomy was followed later by pulmonary surgery. The masses removed at laparotomy consisted of necrotic and fibrotic tissue in 25.8%, mature teratoma in 61.3% and viable cancer was detected in 12.9% cases. In contrast, resected pulmonary masses consisted of necrotic and fibrotic tissue in 42.1%, mature teratoma in 26.3% and viable cancer in 31.6%. The total therapeutic outcome in 220 pts was as follows: 159 pts (72.3%) are alive and free of disease. Four pts (1.8%) live with the disease and 57 pts (25.9%) died from the disease. On the basis of their results the authors advocate: 1. Primary chemotherapy should be indicated for IIA and IIB stages of nonseminomatous testicular tumours. 2. Surgical exploration should be reserved for pts with residual abdominal and pulmonary masses bigger than 15 mm in diameter. 3. Operation should include a complete removal of all residual masses. 4. No reliable criteria indicating necrotic and fibrotic changes preoperatively were identified. (Tab. 4, Ref. 17.)


Growth of adolescents in Mali (West Africa).
This report describes the growth curve during adolescence in the rural area of Mali (West Africa). The authors measured the height, weight, and arm circumference in a sample of 1255 subjects over 14 years and below 20 years of age during a 2-year cross-sectional study (1984-1985). The comparison of the trend of these criteria with French data suggests that Malian adolescents present a different growth curve to those observed in industralized countries.


Clostridium butyricum MIYAIRI 588 alleviates periodontal bone loss in mice with diabetes mellitus.
The gut microbiota is a bridge linking periodontitis and systemic diseases, such as diabetes mellitus (DM). The probiotic Clostridium butyricum MIYAIRI 588 (CBM588) is reportedly an effective therapeutic approach for gut dysbiosis. Here, in a mouse model, we explored the therapeutic effect of CBM588 on periodontal bone destruction in DM and DM-associated periodontitis (DMP), as well as the underlying mechanism. Micro-computed tomography revealed that DM and DMP both aggravated periodontal bone destruction, which was alleviated by intragastric supplementation with CBM588. Moreover, 16S rRNA sequencing and untargeted metabolite analysis indicated that CBM588 ameliorated DMP-triggered dysbiosis and led to reduced oxidative stress associated with elevated 4-hydroxybenzenemethanol (4-HBA) in serum. Furthermore, in vitro and in vivo experiments found that the metabolite 4-HBA promoted nuclear factor erythroid 2-related factor 2 (Nrf2) signaling activation and modulated the polarization of macrophages, thus ameliorating inflammatory bone destruction in DMP. Our study demonstrates the protective effects of CBM588 in DM-induced mice, with and without ligature-induced periodontitis. The mechanism involves regulation of the gut microbiota and restoration of the integrity of the gut barrier to alleviate oxidative damage by elevating serum 4-HBA. This study suggests the possibility of CBM588 as a therapeutic adjuvant for periodontal treatment in diabetes patients.


Experience with the resection of parapharyngeal cancers via the infratemporal fossa approach.
Between 1982 and 1984, a modified infratemporal fossa approach was used to resect cancers with extensive primary or secondary involvement of the infratemporal fossa and parapharynx in 10 patients. Nine patients exhibited persistent or recurrent disease of the upper aerodigestive tract and posterior cranial fossa following planned, curative-intent therapy; the remaining patient had a carcinoma ex-pleomorphic adenoma of the deep lobe of the parotid gland with a significant infratemporal fossa extension. Considered to have "unresectable" tumors, by traditional methods, 7 of the 10 patients underwent an en-bloc resection of their lesions with tumor-free margins. Tumor was present at the margins of the specimens in the other 3 patients. Two of the 10 patients died early in the postoperative period of medical complications. Another died 5 months postoperatively of a tumor-induced internal carotid artery rupture at the level of the foramen lacerum. A fourth patient died of his disease 6 months following his resection. One patient is alive, but has metastatic meningioma 2 years after surgery. The 5 remaining patients are without evidence of disease, with a mean follow up of 2 years. Indications for and refinements of the operative technique, particularly those related to the repair of such extensive ablative defects, are outlined on the basis of this early experience.


[Transplantation of animal organs (xenotransplantation). Status and future development].
The increasing shortage of human organs for allotransplantation has created intense interest for xenotransplantation. Pig organs have been suggested as suitable grafts for man. However, the immediate problem associated with discordant xenotransplantation is hyperacute rejection. Hyperacute rejection occurs because antibodies become attached to the endothelial cells of the xenotransplant organ, which activates the complement system, attracts platelets etc. This results in endothelial cell damage, oedema, loss of vascular integrity and thus rapid destruction of the graft. Organs from non-human primates often do not cause hyperacute rejection, but will probably not be acceptable donors. Transgenic pigs have been breeded which contain the genes for the human regulators of complement activation. It appears that insertion of such genes into pigs serves to prevent aggressive hyperacute rejection. Understanding the mechanisms of rejection gives cause for optimism. However, there are still many problems to evaluate and solve. One potential danger associated with xenotransplantation is the transmission of infectious disease from animals to man. Other problems relate to physiological adaptation of the xenograft. The legal, ethical, religious and economic aspects must also be taken into consideration. Despite the potential problems, it is likely that a xenotransplant of some kind will soon be attempted in man.


Guided self-help cognitive behavioural therapy for depression in primary care: a randomised controlled trial.
Access to Cognitive behavioural therapy (CBT) for depression is limited. One solution is CBT self-help books. Trial Objectives: To assess the impact of a guided self-help CBT book (GSH-CBT) on mood, compared to treatment as usual (TAU). GSH-CBT will have improved mood and knowledge of the causes and treatment of depression compared to the control receiving TAUGuided self-help will be acceptable to patients and staff. Adults attending seven general practices in Glasgow, UK with a BDI-II score of ≥14. 141 randomised to GSH-CBT and 140 to TAU. RCT comparing 'Overcoming Depression: A Five Areas Approach' book plus 3-4 short face to face support appointments totalling up to 2 hours of guided support, compared with general practitioner TAU. The BDI (II) score at 4 months. Numbers analysed: 281 at baseline, 203 at 4 months (primary outcome), 117 at 12 months. Mean BDI-II scores were lower in the GSH-CBT group at 4 months by 5.3 points (2.6 to 7.9, p<0.001). At 4 and 12 months there were also significantly higher proportions of participants achieving a 50% reduction in BDI-II in the GSH-CBT arm. The mean support was 2 sessions with 42.7 minutes for session 1, 41.4 minutes for session 2 and 40.2 minutes of support for session 3. Adverse effects/Harms: Significantly less deterioration in mood in GSH-CBT (2.0% compared to 9.8% in the TAU group for BDI-II category change). Weaknesses: Our follow-up rate of 72.2% at 4 months is better than predicted but is poorer at 12 months (41.6%). In the GSH-CBT arm, around 50% of people attended 2 or fewer sessions. 22% failed to take up treatment. GSH-CBT is substantially more effective than TAU. Controlled-Trials.com ISRCTN13475030.


Relationships Between Measures of the Ability to Perform Vision-Related Activities, Vision-Related Quality of Life, and Clinical Findings in Patients With Glaucoma.
To our knowledge, few studies have combined an objective measure of vision-related performance (VRP) and subjective measures of vision-related quality of life (VRQoL) with clinically related visual parameters, particularly in a large, prospective, cohort study setting. To examine the relationships between clinical visual assessments and both a VRP and 2 self-reported VRQoL measurements. Patients (N = 161) with moderate-stage glaucoma recruited from the Glaucoma Service at Wills Eye Hospital, Philadelphia, Pennsylvania, were enrolled from May 2012 to May 2014 in an ongoing prospective, 4-year longitudinal observational study. This report includes cross-sectional results from the baseline visit. Patients received a complete ocular examination, automated visual field (VF) test and Cirrus optical coherence tomographic scan. Contrast sensitivity was measured with the Pelli-Robson and the Spaeth-Richman Contrast Sensitivity (SPARCS) tests. Vision-related performance was assessed by the Compressed Assessment of Ability Related to Vision (CAARV) test. Vision-related QoL was assessed by the National Eye Institute Visual Function Questionnaire 25 (NEI-VFQ-25) and a modified Glaucoma Symptom Scale (MGSS). Correlations between clinical measures and CAARV, NEI-VFQ-25, and MGSS scores. A total of 161 patients were enrolled in the study. The strongest correlation was found between SPARCS score in the better eye and total CAARV score (r = 0.398; 95% CI, 0.235-0.537; P < .001). The CAARV score also correlated with the Pelli-Robson score (r = 0.353; 95% CI, 0.186-0.499; P = .001), VF mean deviation (r = 0.366; 95% CI, 0.200-0.510; P < .001), and VA (r = -0.326, 95% CI = -0.476 to -0.157; P = .003) in the better eye. There were more statistically significant correlations between contrast sensitivity tests and VF mean deviation with VRQoL measurements than with other clinical measures (visual acuity, intraocular pressure, Disc Damage Likelihood Scale, and mean retinal nerve fiber layer thickness). The MGSS scores were lower (worse) in women compared with men (P = .03 for binocular, P = .01 for better eye, and P = .05 for the worse eye). Structural measures (eg, Disc Damage Likelihood Scale, and retinal nerve fiber layer thickness) were generally not informative with respect to VRP or VRQoL. Contrast sensitivity tests and VF mean deviation were associated with both objective measures of the ability to act and subjective measurements of VRQoL. The strongest correlation was between SPARCS score (contrast sensitivity) in the better eye and total CAARV score. Therefore, measurement of contrast sensitivity should be considered when evaluating patients' VRQoL. The results of this study were limited by the patient population and apply only within the bounds of the tested cohort.


Rho kinase activation and ROS production contributes to the cooling enhanced contraction in cutaneous equine digital veins.
A decrease in environmental temperature can directly affect the contractility of cutaneous vasculature, mediated in part by alpha(2)-adrenoceptors. Most of the cellular mechanisms underlying the cooling-enhanced contractility to alpha(2)-adrenoceptor agonists have been reported in cutaneous arteries but little information is available on cutaneous veins. To investigate the cellular mechanisms associated with the cooling-enhanced contraction to UK-14304 (alpha(2)-adrenoceptor agonist), isolated equine digital veins (EDVs) were studied at 30 degrees C and 22 degrees C. The effects of inhibitors were studied on the contractile response to UK-14304 (0.1 microM). The cooling-enhanced responses were inhibited by Rho kinase inhibitors [maximum response to UK-14304 95.2 +/- 8% of response to depolarizing Krebs solution (DKS) in control vessels cooled to 22 degrees C, compared with 31.4 +/- 6% in the presence of fasudil 1 microM and 75.8 +/- 6% with Y-27632 0.1 microM] and the effects of these inhibitors were considerably less at 30 degrees C (control response 56.4 +/- 5% of DKS; 34.9 +/- 6% with fasudil 1 microM and 50.6 +/- 9% with Y-27632 0.1 microM). Furthermore, Western blotting showed that one of the downstream targets for Rho kinase activity, ezrin/radixin/moesin, was phosphorylated after cooling and reduced by fasudil (1 microM) only at 22 degrees C. The activation of protein kinase C contributed to the contractile response, but predominantly at 30 degrees C (maximum response 82.3 +/- 9% of DKS for control; 57.7 +/- 10% in the presence of chelerythrine 10 microM) with no significant effect at 22 degrees C. The reduction of the response at 22 degrees C by antioxidants, rotenone (14% reduction), and tempol (21% reduction) suggested the contribution of reactive oxygen species (ROS). No evidence was obtained to support the participation of tyrosine kinase. These data demonstrate that Rho kinase activation and the production of ROS contributes to the cooling-enhanced contraction in these cutaneous digital veins.


Management of immune thrombocytopenia: 2022 update of Korean experts recommendations.
Despite the availability of therapies to treat patients with immune thrombocytopenia (ITP), there is currently little data from randomized trials to assist clinicians in managing patients. The evidence-based guidelines of the Korean Society of Hematology Aplastic Anemia Working Party (KSHAAWP) are intended to support patients and physicians in the management of ITP. Experts from the KSHAAWP discussed and described this guideline according to the current treatment situation for ITP in Korea and finalized the guidelines. The expert panel recommended the management of ITP in adult and pediatric patients with newly diagnosed, persistent, and chronic disease refractory to first-line therapy with minor bleeding. Management approaches include observation and administration of corticosteroids, intravenous immunoglobulin, anti-D immunoglobulin, and thrombopoietin receptor agonists. Currently, evidence supporting strong recommendations for various management approaches is lacking. Therefore, a large focus was placed on shared decision-making, especially regarding second-line treatment.


Association of QT interval with blood pressure in 80-year-old subjects.
Few data are available on the association between the prolonged heart rate-adjusted QT (QTc) interval and high blood pressure in elderly individuals, particularly in subjects over 80 years old. The aim of the present study was to determine the association between the QTc interval and blood pressure in 80-year-old subjects. This study was part of the 8020 Data Bank Survey, which was designed to collect the baseline data of systemic and dental health conditions in 80-year-old subjects. We studied the cross-sectional association of the QTc interval with blood pressure in 642 Japanese (257 men and 385 women), all 80 years old. Mean systolic blood pressure (SBP) rose from 146.0 mmHg in the first quartile of QTc interval to 149.1 mmHg in the second, 154.6 mmHg in the third, and 152.3 mmHg in the fourth quartile (test for trend, p=0.008). Mean diastolic blood pressure (DBP) also rose from 76.9 mmHg in the first quartile of QTc interval to 77.7 mmHg in the second, 81.8 mmHg in the third, and 79.0 mmHg in the fourth quartile (test for trend, p=0.003). We performed multiple regression analysis, controlling for factors known to influence the QTc intervals-e.g., SBP, heart rate, sex, and left ventricular hypertrophy assessed by the voltage amplitudes recorded in the precordial leads of the electrocardiogram. The association between the QTc interval and SBP was highly statistically significant in all analyses. These results show that SBP by itself may influence the QTc interval in very old subjects.


Spontaneous sublingual haematoma: a rare entity.
Sublingual haematoma, also known as pseudo-Ludwig phenomenon, is an entity commonly described in patients on anticoagulation therapy. Spontaneous sublingual haematoma is a rare subtype. It is thought to be due to aneurismal changes in the facial or lingual arteries, occurring mostly in the elderly hypertensive population. Two case reports of spontaneous sublingual haematomas have been reported so far in the literature. Both cases were elderly patients and presented with a sublingual mass and elevation of the floor of the mouth. One patient was intubated whereas the other underwent an emergency tracheotomy. The authors present the third case of spontaneous sublingual haematoma that was afebrile and had a sudden onset of sore throat that progressed to dyspnoea and required a life-saving tracheotomy. The patient recovered quickly postoperatively and could be discharged home in a couple of days and was decannulated in 1 week. No recurrence of the haematoma was noticed on follow-up after 1 year.


Annual Research Review: Sex, gender, and internalizing conditions among adolescents in the 21st century - trends, causes, consequences.
Internalizing conditions of psychopathology include depressive and anxiety disorders; they most often onset in adolescence, are relatively common, and contribute to significant population morbidity and mortality. In this research review, we present the evidence that internalizing conditions, including depression and anxiety, as well as psychological distress, suicidal thoughts and self-harm, and fatal suicide, are considerably increasing in adolescent populations across many countries. Evidence indicates that increases are currently greatest in female adolescents. We present an epidemiological framework for evaluating the causes of these increases, and synthesize research on whether several established risk factors (e.g., age of pubertal transition and stressful life events) and novel risk factors (e.g., digital technology and social media) meet conditions necessary to be plausible causes of increases in adolescent internalizing conditions. We conclude that there are a multitude of potential causes of increases in adolescent internalizing conditions, outline evidence gaps including the lack of research on nonbinary and gender nonconforming populations, and recommend necessary prevention and intervention foci from a clinical and public health perspective.


[Review on the evaluation research of the effects of smoke-free legislations on cardiovascular diseases].
A substantial amount of data suggested that exposure to secondhand smoke (SHS) could cause cardiovascular diseases and acute coronary events in nonsmoking adults. In order to protect the public from SHS, more and more countries and regions across the world have enacted and implemented smoke-free legislations. Developed countries, such as USA, Canada, the Great Britain, Ireland, Italy and Spain, have carried out many studies to analyze the effects of smoke-free legislations on the hospital admission, incidence and mortality of cardiovascular diseases with the purpose of confirming the health benefits of the smoking ban and promoting the conduct of the ban. We searched PubMed, EMBASE, Web of Science, Cochrane Library,China National Knowledge Infrastructure(CNKI),WANFANG databaseto summarize the study designs, evaluating indicators, statistical methods and results of these studies to provide reference for evaluating smoke-free legislations in inland cities in China.


Photoresponsive peptoid oligomers bearing azobenzene side chains.
N-Substituted glycine peptoid oligomers were synthesized to incorporate a photoresponsive azobenzene side chain. The ability of this side chain to undergo reversible photoisomerization was established, and the cis- to trans-azobenzene thermal isomerization of this side chain was investigated. Circular dichroism studies indicated that trans- to cis-azobenzene isomerization does not significantly alter the backbone conformation in a series of peptoids thought to have well-defined structures.


Meso-2,3-dimercaptosuccinic acid in the diagnosis and treatment of lead poisoning.
Lead poisoning remains one of the hazards of industrialized civilization. CaNa2 EDTA and dimercaprol, the usual therapeutic measures, have many side effects and can be given by parenteral route alone. The authors present a case of chronic lead poisoning caused by ingestion of contaminated flour ground in a primitive flour mill. The diagnosis was confirmed by the CaNa2 EDTA provocative test. Dimercaptosuccinic acid (DMSA) was given orally as a further provocation and resulted in an 11-fold increase in urinary lead excretion. A 5-day course of treatment with DMSA was instituted, during which symptoms abated, urinary lead excretion increased and the blood lead level decreased. No side effects were noticed. There has been no relapse over several months of follow-up. The authors conclude that the oral use of DMSA is effective, safe and convenient both as a provocative test in establishing the diagnosis of lead poisoning and as a therapeutic tool.


Role of social environment and social clustering in spread of opinions in coevolving networks.
Taking a pragmatic approach to the processes involved in the phenomena of collective opinion formation, we investigate two specific modifications to the coevolving network voter model of opinion formation studied by Holme and Newman [Phys. Rev. E 74, 056108 (2006)]. First, we replace the rewiring probability parameter by a distribution of probability of accepting or rejecting opinions between individuals, accounting for heterogeneity and asymmetric influences in relationships between individuals. Second, we modify the rewiring step by a path-length-based preference for rewiring that reinforces local clustering. We have investigated the influences of these modifications on the outcomes of simulations of this model. We found that varying the shape of the distribution of probability of accepting or rejecting opinions can lead to the emergence of two qualitatively distinct final states, one having several isolated connected components each in internal consensus, allowing for the existence of diverse opinions, and the other having a single dominant connected component with each node within that dominant component having the same opinion. Furthermore, more importantly, we found that the initial clustering in the network can also induce similar transitions. Our investigation also indicates that these transitions are governed by a weak and complex dependence on system size. We found that the networks in the final states of the model have rich structural properties including the small world property for some parameter regimes.


Leiomyosarcoma of the renal vein: analysis of outcome and prognostic factors in the world case series of 67 patients.
Leiomyosarcoma is a rare malignant mesenchymal tumour. Some cases of leiomyosarcoma of the renal vein (LRV) have been reported in the literature, but no analysis of data and search for prognostic factors have been done so far. The aim of this review was to describe the LRV, to analyse overall survival (OS), local recurrence free survival (LRFS) and distant metastases free survival (DMFS) in LRV world case series and to identify significant predictors of OS, LRFS and DMFS. Cases from the literature based on PubMed search and a case from our institution were included. Sixty-seven patients with a mean age of 56.6 years were identified; 76.1% were women. Mean tumour size was 8.9 cm; in 68.7% located on the left side. Tumour thrombus extended into the inferior vena cava lumen in 13.4%. All patients but one underwent surgery (98.5%). After a median follow up of 24 months, the OS was 79.5%. LRFS was 83.5% after a median follow up of 21.5 months and DMFS was 76.1% after a median follow up of 22 months. Factors predictive of OS in univariate analysis were surgical margins, while factors predictive of LRFS were inferior vena cava luminal extension and grade. No factors predictive of DMFS were identified. In multivariate analysis none of the factors were predictive of OS, LRFS and DMFS. Based on the literature review and presented case some conclusions can be made. LRV is usually located in the hilum of the kidney. It should be considered in differential diagnosis of renal and retroperitoneal masses, particularly in women over the age 40, on the left side and in the absence of haematuria. Core needle biopsy should be performed. Patients should be managed by sarcoma multidisciplinary team. LRV should be surgically removed, with negative margins.


Localized redox relays as a privileged mode of cytoplasmic hydrogen peroxide signaling.
Hydrogen peroxide (H2O2) is a key signaling agent. Its best characterized signaling actions in mammalian cells involve the early oxidation of thiols in cytoplasmic phosphatases, kinases and transcription factors. However, these redox targets are orders of magnitude less H2O2-reactive and abundant than cytoplasmic peroxiredoxins. How can they be oxidized in a signaling time frame? Here we investigate this question using computational reaction-diffusion models of H2O2 signaling. The results show that at H2O2 supply rates commensurate with mitogenic signaling a H2O2 concentration gradient with a length scale of a few tenths of μm is established. Even near the supply sites H2O2 concentrations are far too low to oxidize typical targets in an early mitogenic signaling time frame. Furthermore, any inhibition of the peroxiredoxin or increase in H2O2 supply able to drastically increase the local H2O2 concentration would collapse the concentration gradient and/or cause an extensive oxidation of the peroxiredoxins I and II, inconsistent with experimental observations. In turn, the local concentrations of peroxiredoxin sulfenate and disulfide forms exceed those of H2O2 by several orders of magnitude. Redox targets reacting with these forms at rate constants much lower than that for, say, thioredoxin could be oxidized within seconds. Moreover, the spatial distribution of the concentrations of these peroxiredoxin forms allows them to reach targets within 1 μm from the H2O2 sites while maintaining signaling localized. The recruitment of peroxiredoxins to specific sites such as caveolae can dramatically increase the local concentrations of the sulfenic and disulfide forms, thus further helping these species to outcompete H2O2 for the oxidation of redox targets. Altogether, these results suggest that H2O2 signaling is mediated by localized redox relays whereby peroxiredoxins are oxidized to sulfenate and disulfide forms at H2O2 supply sites and these forms in turn oxidize the redox targets near these sites.


Environment of the active site region of RseP, an Escherichia coli regulated intramembrane proteolysis protease, assessed by site-directed cysteine alkylation.
Regulated intramembrane proteolysis (RIP) plays crucial roles in both prokaryotic and eukaryotic organisms. Proteases for RIP cleave transmembrane regions of substrate membrane proteins. However, the molecular mechanisms for the proteolysis of membrane-embedded transmembrane sequences are largely unknown. Here we studied the environment surrounding the active site region of RseP, an Escherichia coli S2P ortholog involved in the sigma(E) pathway of extracytoplasmic stress responses. RseP has two presumed active site motifs, HEXXH and LDG, located in membrane-cytoplasm boundary regions. We examined the reactivity of cysteine residues introduced within or in the vicinity of these two active site motifs with membrane-impermeable thiol-alkylating reagents under various conditions. The active site positions were inaccessible to the reagents in the native state, but many of them became partially modifiable in the presence of a chaotrope, while requiring simultaneous addition of a chaotrope and a detergent for full modification. These results suggest that the active site of RseP is not totally embedded in the lipid phase but located within a proteinaceous structure that is partially exposed to the aqueous milieu.


Genetic and environmental contributions to stability and change in children's internalizing and externalizing problems.
To estimate genetic, shared environmental, and nonshared environmental contributions to stability and change in internalizing and externalizing problems. Maternal Child Behavior Checklist ratings were obtained for 3,873 twin pairs at age 3 and 1,924 twin pairs at age 7. For 1,575 twin pairs, ratings were available at both ages. For Internalizing/Externalizing ratings, genetic, shared, and nonshared environmental factors explained about 59/51%, 10/30%, and 31/19% of the variance at age 3, and 40/52%, 31/32%, and 29/16% of the variance at age 7. The phenotypic correlation of r = 0.38/0.54 between problems assessed at 3 and 7 years of age was explained for 66/55% by genetic factors, for 23/37% by shared environmental factors, and for 11/8% by nonshared environmental factors. The genetic, shared environmental, and nonshared environmental correlations between ages 3 and 7 were 0.51/0.57, 0.47/0.66, and 0.13/0.24, respectively. Genetic and shared environmental factors were most important for the stability of Internalizing and Externalizing Problems between ages 3 and 7. Nonshared environmental factors were mainly age-specific. For Internalizing Problems, shared environment may become more important from early to middle childhood.


Pancreatic trophism following colectomy in rats: the potential role of gastrointestinal hormones.
It has been shown that the large bowel contains substances with a potential to inhibit exocrine pancreatic function. Following large bowel removal in rats, there is an increase of pancreatic weight, digestive enzyme concentration, and secretion capacity in vitro. To evaluate the role of various GI hormones in the exocrine pancreatic adaptation following colectomy, we measured plasma cholecystokinin (CCK), neurotensin, glucagon, and insulin after meal stimulation. The test meal was applied via a transabdominal gastric tube in eight colectomized Wistar rats after a median of 18 days following surgery. Ten rats with a gastric tube without previous bowel surgery served as controls. After large bowel removal, there was impaired glucose tolerance and attenuated plasma insulin secretion. Baseline plasma glucagon levels were increased after colon removal, whereas the total postprandial glucagon release was decreased. Baseline and postprandial neurotensin values were comparable in both the experimental and control animals. Baseline and postprandial CCK plasma levels were intensely increased in the colectomized rats. It is assumed that the baseline and postprandial CCK pattern in rats after subtotal colectomy is responsible for exocrine pancreatic adaptation.


Are mammography image acquisition factors, compression pressure and paddle tilt, associated with breast cancer detection in screening?
To assess the associations between objectively measured mammographic compression pressure and paddle tilt and breast cancer (BC) detected at the same ("contemporaneous") screen, subsequent screens, or in-between screens (interval cancers). Automated pressure and paddle tilt estimates were derived for 80,495 mammographic examinations in a UK population-based screening programme. Adjusted logistic regression models were fitted to estimate the associations of compression parameters with BC detected at contemporaneous screen (777 cases).Nested case-control designs were used to estimate associations of pressure and tilt with: (a) interval cancer (148 cases/625 age-matched controls) and (b) subsequent screen-detected cancer (344/1436), via conditional logistic regression. Compression pressure was negatively associated with odds of BC at contemporaneous screen (odds ratio (OR) for top versus bottom third of the pressure distribution: 0.74; 95% CI 0.60, 0.92; P-for-linear-trend (Pt) = 0.007). There was weak evidence that moderate pressure at screening was associated with lower odds of interval cancer (OR for middle versus bottom third: 0.63; 95% CI 0.38, 1.05; p = 0.079), but no association was found between pressure and the odds of BC at subsequent screen. There was no evidence that paddle tilt was associated with the odds of contemporaneous, subsequent screen or interval cancer detection. Findings are consistent with compression pressure, but not paddle tilt, affecting the performance of mammographic screening by interfering with its ability to detect cancers. Inadequate or excessive compression pressure at screening may contribute to a reduced ability to detect cancers, resulting in a greater number of interval cancer cases.


Zinc supplementation in very-low-birth-weight infants.
Inadequate zinc intake may lead to poor growth and developmental outcome in very-low-birth-weight (VLBW; < 1,500 g) infants. Fifty-two infants (mean birth weight, 1,117 +/- 287 g; mean gestational age, 29 +/- 2.9 weeks) were randomly allocated to two groups. SUPP infants received a regular term formula plus zinc supplements (4.4 mg/L; final content, 11 mg/L); PLAC infants received the same formula plus placebo (final content, 6.7 mg/L). Infants started their formula at 1,853 +/- 109 g and consumed the formula for 6 months. All subjects were evaluated at 3, 6, 9, and 12 +/- 0.75 months corrected-for-gestational-age. At each evaluation, weight, length, and head circumference were measured, a Griffiths developmental assessment was performed, and a blood sample was taken. Higher plasma zinc levels (p < 0.05) were found in the SUPP group at 1 and 3 months, and improved linear growth velocity was found in the SUPP group over the study period for the whole group as well as for girls alone. Maximum motor development scores were higher (p = 0.018) in the SUPP (98 +/- 10) than the PLAC (90 +/- 8) group, indicating that increased zinc intake in early infancy may be beneficial to VLBW infants.


Albuterol aerosol delivered via metered-dose inhaler to intubated pediatric models of 3 ages, with 4 spacer designs.
To determine the amount of albuterol, in various particle size ranges, delivered from a hydrofluoroalkane-propelled metered-dose inhaler (Airomir) in 3 models of pediatric intubation (ages 8 months, 4 years, and 16 years) using 4 types of aerosol reservoir: 3 spacers (ACE, AeroChamber HC MV, metal NebuChamber without 1-way valve) and 1 holding chamber (metal NebuChamber with 1-way valve). Five reservoirs of each type were tested with albuterol sulfate delivered via metered-dose inhaler that delivers 100 microg of albuterol per actuation. Each reservoir was connected to an endotracheal tube (ETT) that corresponded to the given patient age (8 months = 4 French; 4 years = 5 French; 16 years = 7.5 French) and to a valved system that allowed connection of the ETT to a cascade impactor. Simulated tidal volumes representative of children of the given ages were passed through the reservoir. Both the cascade impactor and the ETT were enclosed within a 100% humidity, 37 degrees C environment. For the total amount of albuterol inhaled onto the impactor, and both the 1.1-4.7 microm and 1.1-3.3 microm inhaled fine-particle fractions, the NebuChamber-with-valve showed significantly greater drug delivery than the NebuChamber-without-valve, the AeroChamber HC MV, or the ACE (p < 0.001). Among the reservoirs without valves the NebuChamber showed significantly greater delivery than the AeroChamber HC MV or ACE (p < 0.001) for total drug deposition and for both the 1.1-4.7 microm and 1.1-3.3 microm fine-particle fractions. These results were consistent over all age groups. The AeroChamber HC MV had significantly greater delivery (total deposition) than the ACE (p < 0.001), except in the 4-year-old model. There were no significant differences between the AeroChamber HC MV and the ACE for either the 1.1-4.7 microm or the 1.1-3.3 microm fine-particle fraction. An aerosol reservoir with 1-way valve positioned between the spacer and the ETT improved the amount of inhaled albuterol 300-900%, compared to the other reservoirs.


Severe acute renal failure due to tubulointerstitial nephritis, pancreatitis, and hyperthyroidism in a patient during rifampicin therapy.
It is well known that rifampicin can cause nephrotoxicity. Rifampicin-related pancreatitis and hyperthyroidism are rarely reported in the same patient in the presence of tubulointerstitial nephritis. Reported herein is the medical management of a patient with hemolytic anemia, acute renal failure, pancreatitis, and hyperthyroidism during with rifampicin therapy. A 50-year-old man was admitted to the hospital owing to abdominal colic and acute renal failure. He was treated with 2 courses of tetracycline-rifampicin for brucellosis 3 weeks and 4 months prior to admission. Physical examination showed blood pressure of 130/70 mm Hg, pulmonary crackles, and edema. Laboratory findings are detailed in the case report. Findings of abdominal ultrasonography suggested edematose pancreatitis and thyroid ultrasonography showed several solid nodules. Renal biopsy showed tubulointerstitial nephritis. Although rifampicin-related tubulointerstitial nephritis and acute renal failure are not uncommon during rifampicin therapy, the convergence of hyperthyroidism, pancreatitis, tubulointerstitial nephritis, and acute renal failure rarely presents in the same patient. Although pancreatitis, tubulointerstitial nephritis, and acute renal failure were ameliorated with corticoid therapy within 2 months, hyperthyroidism continued and required antithyroid therapy. In conclusion, rifampicin may trigger hyperthyroidism in patients with goiter.


Associative olfactory learning in the desert locust, Schistocerca gregaria.
Locusts can learn associations between olfactory stimuli and food rewards, and use the acquired memories to choose between foods according to their nutrient requirements. They are a model system for both the study of olfactory coding and insect nutritional regulation. Previous studies have used operant paradigms for conditioning freely moving locusts, restricting the study of the neural mechanisms underlying the acquisition of olfactory memories, which requires restrained preparations for electrophysiological recordings. Here we present two complementary paradigms for the classical conditioning of olfactory memories in restrained desert locusts (Schistocerca gregaria). These paradigms allow precise experimental control over the parameters influencing learning. The first paradigm is based on classical (Pavlovian) appetitive conditioning. We show that opening of the maxillary palps can be used as a measure of memory acquisition. Maxillary palp opening in response to odour presentation is significantly higher in locusts trained with paired presentation of an odour and a food reward than in locusts trained either with unpaired presentation of food and odour or the odour alone. The memory formed by this conditioning paradigm lasts for at least 24 h. In the second paradigm, we show that classical conditioning of an odour memory in restrained locusts influences their decisions in a subsequent operant task. When locusts that have been trained to associate an odour with a food reward are placed in a Y-maze, they choose the arm containing that odour significantly more often than naïve locusts. A single conditioning trial is sufficient to induce a significant bias for that odour for up to 4 h. Multiple- and block-trial training induce a significant bias that lasts at least 24 h. Thus, locusts are capable of forming appetitive olfactory memories in classical conditioning paradigms and can use these memories to modify their decisions.


PCR for the identification of methicillin-resistant Staphylococcus aureus (MRSA) strains using a single primer pair specific for SCCmec elements and the neighbouring chromosome-borne orfX.
The chromosomal location of the SCCmec elements containing mecA allows the identification of methicillin-resistant Staphylococcus aureus (MRSA) strains by PCR amplification of a sequence covering the right junction of the SCCmec elements and the adjacent chromosomal region encoding the species-specific ORFX. MRSA strains can be identified specifically using one forward primer, with only one or two mismatches, targeting the SCCmec elements of different types, and one reverse primer targeting the orfX region.


Artificial Intelligence in Ventricular Arrhythmias and Sudden Death.
Sudden cardiac arrest due to lethal ventricular arrhythmias is a major cause of mortality worldwide and results in more years of potential life lost than any individual cancer. Most of these sudden cardiac arrest events occur unexpectedly in individuals who have not been identified as high-risk due to the inadequacy of current risk stratification tools. Artificial intelligence tools are increasingly being used to solve complex problems and are poised to help with this major unmet need in the field of clinical electrophysiology. By leveraging large and detailed datasets, artificial intelligence-based prediction models have the potential to enhance the risk stratification of lethal ventricular arrhythmias. This review presents a synthesis of the published literature and a discussion of future directions in this field.


Effects of corn silage derived from a genetically modified variety containing two transgenes on feed intake, milk production, and composition, and the absence of detectable transgenic deoxyribonucleic acid in milk in Holstein dairy cows.
The objectives were to compare the chemical composition, nutritive value, feed intake, milk production and composition, and presence in milk of transgenic DNA and the encoded protein Cry1Ab when corn silages containing 2 transgenes (2GM: herbicide tolerance: mepsps and insect resistance: cry1Ab) were fed as part of a standard total mixed ration (TMR) compared with a near isogenic corn silage (C) to 8 multiparous lactating Holstein dairy cows in a single reversal design study. Cows were fed a TMR ration ad libitum and milked twice daily. Diets contained [dry matter (DM) basis] 45% corn silage, 10% alfalfa hay, and 45% concentrate (1.66 Mcal of net energy for lactation/kg of DM, 15.8% crude protein, 35% neutral detergent fiber, and 4.1% fat). Each period was 28-d long. During the last 4 d of each period, feed intake and milk production data were recorded and milk samples taken for compositional analysis, including the presence of transgenic DNA and Cry1Ab protein. There was no significant difference in the chemical composition between C and 2GM silages, and both were within the expected range (37.6% DM, 1.51 Mcal of net energy for lactation/kg, 8.6% crude protein, 40% neutral detergent fiber, 19.6% acid detergent fiber, pH 3.76, and 62% in vitro DM digestibility). Cows fed the 2GM silage produced milk with slightly higher protein (3.09 vs. 3.00%), lactose (4.83 vs. 4.72%) and solids-not-fat (8.60 vs. 8.40%) compared with C. However, the yield (kg/d) of milk (36.5), 3.5% fat-corrected milk (34.4), fat (1.151), protein (1.106), lactose (1.738), and solids-not-fat (3.094), somatic cell count (log10: 2.11), change in body weight (+7.8 kg), and condition score (+0.09) were not affected by type of silage, indicating no overall production difference. All milk samples were negative for the presence of transgenic DNA from either trait or the Cry1Ab protein. Results indicate that the 2GM silage modified with 2 transgenes did not affect nutrient composition of the silages and had no effect on animal performance and milk composition. No transgenic DNA and Cry1Ab protein were detected in milk.


Platelet-Rich Plasma Versus Corticosteroids in the Treatment of Chronic Plantar Fasciitis: A Systematic Review and Meta-analysis of Prospective Comparative Studies.
Plantar fasciitis is a common cause of heel pain, which often results in a significant limitation of daily activity. The objective of this review was to compare platelet-rich plasma injection (PRP) and corticosteroids injection in the treatment of chronic plantar fasciitis. MEDLINE, PubMed, Web of Science, EMBASE, and the Cochrane library of clinical trials were searched in March 2019. Inclusion criteria were prospective comparative studies reporting functional scores or pain score with a minimum follow-up of 1 month. We excluded studies with patients younger than 18 years and patients with a history of surgical intervention, infection, or rheumatological disease. First author, publication year, sample size, type of intervention, outcome measures, and follow-up period were recorded. The American Orthopaedic Foot & Ankle Society score was significantly better in the PRP group at 6 months' follow-up, with a lower visual analogue scale at 6- and 12-month intervals. The Foot and Ankle Disability Index and Roles-Maudsley score were similar in the 2 treatments at 3 and 6 months. PRP injection in the treatment of chronic plantar fasciitis is considered safe, with a favorable functional outcome and pain control at intermediate and long-term follow-up in comparison to corticosteroid injection.


Potential role of microRNAs in mammalian female fertility.
Since the first evidence for the involvement of microRNAs (miRNAs) in various reproductive processes through conditional knockout of DICER, several studies have been conducted to investigate the expression pattern and role of miRNAs in ovarian follicular development, oocyte maturation, embryo development, embryo-maternal communication, pregnancy establishment and various reproductive diseases. Although advances in sequencing technology have fuelled miRNA studies in mammalian species, the presence of extracellular miRNAs in various biological fluids, including follicular fluid, blood plasma, urine and milk among others, has opened a new door in miRNA research for their use as diagnostic markers. This review presents data related to the identification and expression analysis of cellular miRNA in mammalian female fertility associated with ovarian folliculogenesis, oocyte maturation, preimplantation embryo development and embryo implantation. In addition, the relevance of miRNAs to female reproductive disorders, including polycystic ovary syndrome (PCOS), endometritis and abnormal pregnancies, is discussed for various mammalian species. Most importantly, the mechanism of release and the role of extracellular miRNAs in cell-cell communication and their potential role as non-invasive markers in female fertility are discussed in detail. Understanding this layer of regulation in female reproduction processes will pave the way to understanding the genetic regulation of female fertility in mammalian species.


The effects of ambient NO2 on lung function in primary schoolchildren.
The effect of ambient NO2 on lung function was investigated in a sample of 423 schoolchildren. At each of four locations NO2 was monitored continuously. Over a 6-month period from January to June 1990 two surveys were performed and spirometry recorded each time for each child. Linear regression was used to estimate the effect of NO2 for different time intervals preceding lung function testing. A decrease of NO2 between surveys was significantly associated with a higher forced vital capacity (FVC) at the second survey. For each microgram/m3 NO2 decrease the model predicted an increase in FVC of 1.5 ml [for the 2-hr mean (P < 0.05)] and 3.1 ml [for the 12-hr mean (P < 0.01)]. We conclude that even at NO2 levels below current air-quality standards children demonstrate significant changes in lung function.


Effect of Sonication Time on Magnetorheological Effect for Monomodal Magnetic Elastomers.
The effect of sonication time on the storage modulus and particle morphology for magnetic elastomers was investigated by dynamic viscoelastic measurements and morphological studies. An ultrasonic wave using a homogenizer was irradiated to magnetic liquids containing 70 wt % carbonyl iron, for up to 30 min before cure. SEM photographs revealed that magnetic particles were randomly dispersed in the polyurethane matrix for magnetic elastomers with sonication. A parameter showing nonlinear viscoelasticity for magnetic elastomers with sonication decreased from 0.75 to 0.4, indicating that the aggregations of magnetic particles had been destroyed by the sonication. The storage modulus at 500 mT at the linear viscoelastic regime significantly increased with the irradiation time, reaching saturation after 10 min; this suggests an increase in the number of chains of magnetic particles by sonication, due to the random dispersion of magnetic particles. At high strains, the storage modulus at 500 mT increased by 8.9 kPa by sonication, indicating the number of chains of magnetic particles which were not destroyed by increased sonication. It was also found that the storage modulus for polyurethane elastomers without magnetic particles was not varied by sonication, suggesting that the polyurethane network was not broken. The effect of sonication time on the viscoelastic properties, and on the magnetorheological response for magnetic elastomers, is discussed.


The influence of feeding diets containing different calcium-phosphorous ratios on the laying hen.
Six experiments were conducted to determine the influence of feeding diets containing different calcium-phosphorus (Ca:P)ratios at different times on the laying hen. Feeding a diet containing 0.75% P from 6 a.m. to 11 a.m. and a diet containing 1.50% P from 3 p.m. to 8 p.m. decreased serum calcium in three or four experiments and decreased specific gravity of eggs in four of four experiments, when compared to that of the birds fed the control diet which contained 0.75% P during both time periods. Specific gravity of eggs from hens fed a diet containing 0.75% P from 6 a.m. to 11 a.m. and a diet containing 0.33% P from 3 p.m. to 8 p.m. was increased in three of four experiments. Although the differences are not significant, with one exception, the trend in the experiments indicate that the level of P in the diet and time of intake can influence shell quality and serum calcium. The dietary treatments had no significant influence on feed consumption, egg production or egg weight during any of these experiments. It was postulated that the adverse or beneficial effect on specific gravity of eggs and serum calcium as a result of feeding diets containing various CaP ratios was due to the influence on calcium absorption and/or bone resorption.


Ludwig-Soret effect of aqueous solutions of ethylene glycol oligomers, crown ethers, and glycerol: Temperature, molecular weight, and hydrogen bond effect.
The thermal diffusion, also called the Ludwig-Soret effect, of aqueous solutions of ethylene glycol oligomers, crown ethers, and glycerol is investigated as a function of temperature by thermal diffusion forced Rayleigh scattering. The Soret coefficient, ST, and the thermal diffusion coefficient, DT, show a linear temperature dependence for all studied compounds in the investigated temperature range. The magnitudes and the slopes of ST and DT vary with the chemical structure of the solute molecules. All studied molecules contain ether and/or hydroxyl groups, which can act as acceptor or donor to form hydrogen bonds, respectively. By introducing the number of donor and acceptor sites of each solute molecule, we can express their hydrogen bond capability. ST and DT can be described by an empirical equation depending on the difference of donor minus acceptor sites and the molecular weight of the solute molecule.


Influenza vaccination in patients with suspected egg allergy.
Egg allergy is not necessarily a contraindication to influenza vaccination. For patients with suspected egg allergy, if the clinician determines benefits to outweigh risks, cautionary measures are available that can enhance safe vaccine administration. Batch to batch variability of egg content in extant influenza vaccines necessitates an informed and cautious approach to vaccination of an egg allergic individual. Although patients with egg allergy are likely to develop egg tolerance by late childhood, tolerance to ingestion of "baked egg" products may not predict tolerance to "native egg" proteins present in the influenza vaccine. Even in cases where the skin test to the vaccine is positive, vaccination may still be cautiously administered, if necessary, in a graded dose protocol.


Development of an automated 7-day 96-well Caco-2 cell culture model.
Caco-2 cells are a widely accepted model to predict permeability and absorption of compounds in humans. We built up an automated 96-wellplate Caco-2 permeation model with reduced growth time. Model compounds (metoprolol, ketoprofen, verapamil, naproxen, hydrochlorthiazide), permeability markers (TEER, Lucifer Yellow, D-[1-(14)C]-mannitol) and confocal microscopy were used to assess the utility of our method on Biomek-FX-automation. The confocal imaging data showed that Caco-2 cells formed monolayers when cultured for 7 days with initial cell density of 2.1 x 10(5) cells/cm2. P-Glycoprotein was present in Caco-2 cells, localized on the plasma membrane. Permeation of model compounds was comparable to those obtained from traditional 12-wellplate experiments.


Algorithmic Generation of Grammar Simplification Rules Using Large Corpora.
There is often a discontinuity between patients' literacy level and educational materials. In response, we are developing an online medical text simplification editor. In this paper, we describe generating grammar simplification rules from a large parallel corpus (N=141,500) containing original sentences and their simplified variants. We algorithmically identified grammatical transformations between sentences (N=26,600) and used distributional characteristics in two corpora to select transformations with the broadest application and the least ambiguity. This resulted in a top set of 146 rules. Two experts evaluated 20 representative rules reflecting 4 characteristics (long/short and weak/strong) each with 5 example sentences. Generally, we found that the rules are helpful for guiding simplification. Using a 5-point Likert scale (5=best), stronger rules scored higher for ease of applying (4.11), overall helpfulness (4.40) and usefulness of examples (4.05). Rule length did not affect the expert scores. The grammar simplification rules are being integrated in our text editor.


Dialkylquinonimines validated as in vivo metabolites of alachlor, acetochlor, and metolachlor herbicides in rats.
The oncogenicity of chloroacetanilide herbicides (1-5) is proposed to involve bioactivation to 2,6-dialkylbenzoquinonimines (quinonimines, 9) based on two earlier observations: (1) in vitro conversion of the alachlor (1) metabolite diethylaniline (7Et2) to 2, 6-diethylquinonimine (9Et2) which reacts readily with GSH and (2) induction of sister chromatid exchanges in human lymphocytes for the parent herbicides and their purported 9 metabolites. This hypothesis lacks in vivo evidence for 9 formation which might be provided by analysis of urine and tissue for thiol adducts of 9. Accordingly, two mercapturates (10Et2 and 10Me2) and a cysteine conjugate (11Me2) were prepared by addition of N-acetylcysteine or cysteine to 9Et2 and the 2,6-dimethyl homologue (9Me2). Mercapturate 10Et2 was characterized by HPLC using the urine of rats treated ip with hydroxyaniline 8Et2, and both mercapturate 10Me2 and cysteine conjugate 11Me2 were found in the urine of mice administered hydroxyaniline 8Me2. The mercapturates were then converted to the N, N-dimethyl-2,6-dialkyl-4-methoxy-3-(methylthio)anilines (12Et2, 12EtMe, and 12Me2) by alkaline permethylation, thereby providing a method for analysis of 9-derived thiol adducts in urine and liver as the 12 derivatives by GC/MS with selected ion monitoring. The urine of rats 0-6 h after ip treatment with 1, butachlor (2), acetochlor (3), metolachlor (4), and dimethachlor (5) at 0.74 mmol/kg yields permethylated derivatives which are definitively diagnostic for the 9 intermediates from each of the herbicides in amounts of 3-24-fold above the minimum detectable limit, as well as 1 and 2 orders of magnitude higher from the corresponding anilines (7) and hydroxyanilines (8), respectively. Similar liver analyses reveal tissue thiol adducts of 9 6 h after treatment with 7 and 8 but not with the parent herbicides. The yields of urinary 9 derivatives from the parent herbicides are higher from the 2,6-diethyl series (1 and 2) and the 2-ethyl-6-methyl derivatives (3 and 4) than from the 2, 6-dimethyl analogue (5). These findings provide direct evidence in vivo that quinonimines are metabolites of 1-5 in rats.


Novel Epigenetic Controlling of Hypoxia Pathway Related to Overexpression and Promoter Hypomethylation of TET1 and TET2 in RPE Cells.
CpG methylation of DNA takes part in a specific epigenetic memory that plays crucial roles in the differentiation and abnormality of the cells. The methylation pattern aberration of genomes is affected in three ways, namely DNA methyltransferase (DNMT), ten-eleven translocation (TET), and methyl-binding domain (MBD) proteins. Of these, TET enzymes have recently been demonstrated to be master modifier enzymes in the DNA methylation process. Additionally, recent studies emphasize that not only epigenetic phenomena play a role in controlling hypoxia pathway, but the hypoxia condition also triggers hypomethylation of genomes that may help with the expression of hypoxia pathway genes. In this study, we suggested that TET1 and TET2 could play a role in the demethylation of genomes under chemical hypoxia conditions. Herein, the evaluating methylation status and mRNA expression of mentioned genes were utilized through real-time PCR and methylation-specific PCR (MSP), respectively. Our results showed that TET1 and TET2 genes were overexpressed (P < 0.05) under chemical hypoxia conditions in Retinal Pigment Epithelial (RPE) cells, whereas the promoter methylation status of them were hypomethylated in the same condition. Therefore, chemical hypoxia not only causes overexpression of TET1 and TET2 but also could gradually do promoter demethylation of same genes. This is the first study to show the relationship between epigenetics and the expression of mentioned genes related to hypoxia pathways. Furthermore, it seems that these associations in RPE cells are subjected to chemical hypoxia as a mechanism that could play a crucial role in methylation pattern changes of hypoxia-related diseases such as cancer and ischemia. J. Cell. Biochem. 118: 3193-3204, 2017. © 2017 Wiley Periodicals, Inc.


Reading assessment post head injury: how valid is it?
The purpose of this study was to identify the passage dependency for three reading tests applied to the closed head injured (CHI) population and to determine whether reading the passage resulted in improved accuracy in responding to passage-related questions. Based on the performance of 10 CHI adults, passage dependency was low for all three tests. Only the Reading Comprehension Battery for Aphasia scores improved significantly when the subjects read the passages prior to responding. The results parallel those found with aphasic subjects and raise questions concerning the validity of these tests in assessing communication dysfunction post-CHI.


Dataset on gait patterns in degenerative neurological diseases.
We collected the gait parameters and lower limb joint kinematics of patients with three different types of primary degenerative neurological diseases: (i) cerebellar ataxia (19 patients), (ii) hereditary spastic paraparesis (26 patients), and (iii) Parkinson's disease (32 patients). Sixty-five gender-age matched healthy subjects were enrolled as control group. An optoelectronic motion analysis system was used to measure time-distance parameters and lower limb joint kinematics during gait in both patients and healthy controls.


[Neural control of the peripheral circulation and blood pressure].
In the XIX century Claude Bernard discovered the action of the nervous system on the peripheral circulation. In the first half of the XX century Ewald Hering discovered the baro-receptor and the reflex control of the heart rate and blood pressure. Cowley and Guyton demonstrated that sino-aortic denervation induces persistent changes in the blood pressure in the dog. The autonomic nervous system is mainly responsible for the regulation of the circulation and blood pressure in the short term on a beat to beat basis. It controls the vasomotor tone, the heart rate and the cardiac output. With the advent of non invasive methods that measure the blood pressure on a beat to beat basis (Finapres) and with the methods of measurement of the variability of the blood pressure in the frequency domain (spectral analysis) we can currently measure many variables including heart rate, blood pressure, stroke volume, peripheral resistances and the baroreceptor sensitivity and make some inferences about their control mechanisms. These variables can be measured at rest in the supine position, standing up, during rhythmic breathing and during the Valsalva maneuver. In this article we present a review of the neural control of the blood pressure and heart rate.


Regulation of c-fos and c-jun expression by calcitonin in human breast cancer cells.
Breast cancer cells (BCC) have calcitonin (CT) receptors, yet the action of the hormone on these cells is largely unknown. We found that CT produced a strong and transient time- and dose-dependent increase in c-fos mRNA in BCC lines. This event was prevented by a protein kinase A (PKA) inhibitor, H89. CT alone did not influence the expression of c-jun and of the tissue inhibitors of metalloproteases (timp) -1 and -2 mRNAs; however, it reduced the induction of these mRNAs by the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA), without apparent changes in the half-life of the mRNA (measured for c-jun). Along the same line, CT reduced the c-jun induction and T-47D growth stimulation by epidermal growth factor (EGF) and insulin. These effects were mimicked by forskolin and/or prevented by H89, suggesting that PKA activation was involved. These results indicate that CT modulates in BCC the mRNA levels of two important growth-related early response genes (c-fos and c-jun) and of two other genes (timp-1 and -2) involved in the control of metastatic events.


Right aortic arch with situs solitus frequently heralds a vascular ring.
We hypothesized that a right aortic arch in situs solitus, with or without an associated cardiovascular malformation, is often associated with a vascular ring. From those born in Southern Nevada between March 2012 and March 2017, we identified 50 (3.6 per 10,000 live births) with a right aortic arch and situs solitus. From the 50 patients, 6 did not meet inclusion criteria for further analysis. Of the 44 remaining, 33 (75%) had a vascular ring. Of the 33 with a vascular ring, 26 (79%) occurred with an isolated right aortic arch, and 7 (21%) had an associated cardiovascular malformation. Of the total 44 patients with a right aortic arch in situs solitus, 34 (79%) were diagnosed prenatally. In conclusion, we found a right aortic arch in situs solitus was often associated with a vascular ring. Further, to the best of our knowledge, no previous general population study has demonstrated an equal or higher right aortic arch, prenatal detection rate of 79%.


Internalization mechanisms of brain-derived tau oligomers from patients with Alzheimer's disease, progressive supranuclear palsy and dementia with Lewy bodies.
Tau aggregates propagate in brain cells and transmit to neighboring cells as well as anatomically connected brain regions by prion-like mechanisms. Soluble tau aggregates (tau oligomers) are the most toxic species that initiate neurodegeneration in tauopathies, such as Alzheimer's disease (AD), progressive supranuclear palsy (PSP), and dementia with Lewy bodies (DLB). Exogenous tau aggregates have been shown to be internalized by brain cells; however, the precise cellular and molecular mechanisms that underlie the internalization of tau oligomers (TauO) remain elusive. Using brain-derived tau oligomers (BDTOs) from AD, PSP, and DLB patients, we investigated neuronal internalization mechanisms of BDTOs, including the heparan sulfate proteoglycan (HSPG)-mediated pathway, clathrin-mediated pathway, and caveolae-mediated pathway. Here, we demonstrated that the HSPG-mediated pathway regulates internalization of BDTOs from AD and DLB, while HSPG-mediated and other alternative pathways are involved in the internalization of PSP-derived tau oligomers. HSPG antagonism significantly reduced the internalization of TauO, prevented tau translocation to the endosomal-lysosomal system, and decreased levels of hyperphosphorylated tau in neurons, the well-known contributor for neurofibrillary tangles (NFT) accumulation, degeneration of neurons, and cognitive decline. Furthermore, siRNA-mediated silencing of heparan sulfate (HS)-synthesizing enzyme, exostosin-2, leads to decreased internalization of BDTOs, prevented tau-induced autophagy-lysosomal pathway impairment, and decreased hyperphosphorylated tau levels. Collectively, these findings suggest that HSPG-mediated endocytosis and exostsin-2 are involved in neuronal internalization of TauO and subsequent tau-dependent neuropathology in AD and DLB.


Coronary thrombolysis with human single-chain, urokinase-type plasminogen activator (pro-urokinase) in patients with acute myocardial infarction.
Single-chain, urokinase-type plasminogen activator (pro-urokinase), isolated from the conditioned medium of a human renal adenocarcinoma cell line, was administered to six patients with evolving myocardial infarction of less than 5 hours' duration and angiographically confirmed total occlusion of the infarct-related coronary artery. The agent was infused intravenously at a rate of 40 mg over 60 minutes immediately followed in three patients by intracoronary infusion of 20 mg over 30 minutes. In four of the six patients complete reperfusion was obtained during intravenous administration and in one patient after 20 minutes of intracoronary infusion. One patient did not respond. The infusion caused significant changes in the fibrinogen level and generation of fibrinogen breakdown products in only one patient. Single-chain, urokinase-type plasminogen activator can induce clot-selective coronary thrombolysis in patients with acute myocardial infarction.


LC/QTOF profile and preliminary stability studies of an enriched flavonoid fraction of Cecropia pachystachya Trécul leaves with potential antidepressant-like activity.
There is increasing interest in natural antioxidants that are candidates for the prevention of brain damage occurring in major depressive disorders. Cecropia pachystachya is a tropical tree species of Central and South America and a rich source of polyphenols, particularly flavonoids. The aim of this study was to characterize the flavonoid profile of an enriched flavonoid fraction of C. pachystachya (EFF-Cp) and evaluate the antidepressant-like effects of its acute administration in behavior, cytokine levels, oxidative stress and energy metabolism parameters. The EFF-Cp chemical characterization was performed by HPLC/DAD and LC/QTOF. The antidepressant-like effects were performed by the forced swimming test, splash test and open field test. EFF-Cp revealed 15 flavonoids, including seven new glycosyl flavonoids for C. pachystachya. Quantitatively, EFF-Cp showed isoorientin (43.46 mg/g), orientin (23.42 mg/g) and isovitexin (17.45 mg/g) as major C-glycosyl flavonoids. In addition, EFF-Cp at doses 50 and 100 mg/kg reduced the immobility time in the forced swimming test, without changing the locomotor activity and grooming time. In addition, EFF-Cp was able to prevent the oxidative damage in some brain areas. In conclusion, the results of this study suggest that EFF-Cp exerts antidepressant-like effects with its antioxidant properties.


Tenascin-C produced by oxidized LDL-stimulated macrophages increases foam cell formation through Toll-like receptor-4.
Atherosclerosis is a chronic inflammatory disease in which both innate and adaptive immunity are involved. Although there have been major advances in the involvement of toll-like receptor 4 (TLR4) and CD36 in the initiation and development of this disease, detailed mechanisms remain unknown. Here, we show that tenascin-C (TN-C) can stimulate foam cell formation and this can be inhibited by a TLR4-blocking antibody or CD36 gene silencing. Our results identify TN-C-TLR4 activation as a common molecular mechanism in oxLDL-stimulated foam cell formation and atherosclerosis. In addition, CD36 is the major scavenger receptor responsible for the TN-C-mediated foam cell formation. Taken together, we have identified that TNC produced by oxLDL-stimulated macrophages increases foam cell formation through TLR4 and scavenger receptor CD36.


Performance Evaluation and Clinical Associations of Immunoassays That Detect Antibodies to Negatively Charged Phospholipids Other Than Cardiolipin.
We evaluate the performance characteristics of antiphosphatidylserine (anti-PS), antiphosphatidylinositol (anti-PI), and antiphospholipid mixture (APhL) enzyme-linked immunosorbent assays (ELISAs) compared with anticardiolipin (aCL) and anti-β2 glycoprotein I (anti-β2GPI) in a large group of patients with antiphospholipid (aPL)-related diseases. Serum samples from 548 patients from the Hopkins and Jamaican systemic lupus erythematosus cohorts, the PROMISSE cohort, and the Antiphospholipid Standardization Laboratory were examined for immunoglobulin G (IgG)/immunoglobulin M (IgM) positivity in aCL, anti-β2GPI, anti-PS, anti-PI, and APhL ELISA assays. All IgG assays were associated with one or more thrombotic and/or obstetric manifestations, with an increased risk associated with higher antibody titers. Analytical performance was similar among assays, but IgG assays performed better than IgM counterparts. Increasing titers of APhL, anti-PS, and anti-PI antibodies could indicate an increased risk of thrombotic and/or obstetric aPL-related manifestations. These assays may be promising biomarkers for particular APS manifestations.


Application of stages of change model to adolescents' physical activity in relation to psychological variables.
This study investigated the stages of change in physical activity in relation to psychological variables: self-efficacy, physical self-concept and barriers to physical activity, as well as gender. Furthermore, cluster profiles were created based on psychological variables to be compared with stages of change groups. Data were obtained from a representative sample of adolescents (N.=1618, 14.46±1.28 years) from Aragon region (Spain). The different strata were selected according to the geographical region, environment, type of school, age and gender Males (75.5%) and very active adolescents (96.9% participant boys and 93.5% participants girls) were more prevalent in the action and maintenance groups. We found significant differences in psychosocial variables by stages of change and gender, showing higher scores in active groups for physical self-concept (effects of gender and stages of change: F(1, 1618)=313.82 P<0.001, η²=0.230) and self-efficacy (F(1, 1618)=324.34 P<0.001, η²=0.236), and lower scores in passive groups for intrapersonal (F(1, 1618)=179.73 P<0.001, η²=0.146) and extrapersonal barriers (F(1, 1618)=189.42 P<0.001, η²=0.153). Psychological cluster profiles created with these variables showed significant relationships with stages of change groups (χ²(1)=162,76, P<0.001; Pearson R=0.359 P<0.001). The findings of the research suggest the importance of the study, showing strong determinants of PA, physical self-concept, self-efficacy and perceived barriers, in relation to the intention to change behaviour, not only stage of change model variables. Furthermore, the study can be relevant to designing more effective health promotion programmes in the adolescent population.


The type 1 equilibrative nucleoside transporter regulates ethanol intoxication and preference.
Adenosine is an important mediator of ethanol intoxication. In vitro, ethanol stimulates adenosine signaling by inhibiting the type 1 equilibrative nucleoside transporter (ENT1), whereas chronic ethanol exposure downregulates ENT1. It is not known, however, whether ENT1 is important for ethanol intoxication or consumption in vivo. Here we report that ENT1-null mice show reduced hypnotic and ataxic responses to ethanol and greater consumption of alcohol as compared with their wild-type littermates. These features are associated with a decrease in adenosine tone, as measured indirectly as a reduction in A(1) receptor-mediated inhibition of glutamate excitatory postsynaptic currents (EPSCs) in the nucleus accumbens, leading to increased phosphorylation of CRE-binding protein (CREB) in the striatum. Treatment with an A(1) receptor agonist decreases EPSC amplitude and reduces ethanol consumption in ENT1-null mice. Our results indicate that ENT1 has a physiological role in ethanol-mediated behaviors and suggest that decreased A(1) adenosine receptor function promotes alcohol consumption.


[Screening the effective target sequences of laryngeal carcinoma related gene LCRG1].
To screen the effective target sequences of laryngeal carcinoma related gene LCRG1 using RNAi. PCR site mutation method was used to reconstruct pSuper vector. Five pairs of siRNA sequences designed by siRNA software were annealed and inserted into the reconstructed pSuper vector. The reconstructed pSuper 362,398,432,789,903,and pSuper vectors were transfected into Hela cell lines and selected with the appropriate drugs to get resistant and pool cells, respectively. The colonies were identified by RT-PCR or real-time RT-PCR analysis. The silence effects were observed by cloning formation analysis. pSuper vector was reconstructed to restore Bgl II restriction enzyme sites using PCR mutation. The RT-PCR or real-time RT-PCR Results of pool clones showed 362, 398, and 432 pool clones all had better effects of LCRG1 gene-silence, especially 362 pool clones. The expression level of LCRG1 mRNA of selected 362 group anti-puromycin clones A2 and A5 was decreased. The Results of clone forming efficiency revealed that the cellular proliferation in A2 of 362 group was significantly higher than that of the vector and control Hela cells (P<0.05). The reconstructed pSuper vector is successfully constructed. The 362 group has better gene silence and has 2 effective 362 group anti-clones, suggesting that methodology has important values in studYing the function and molecular mechanism of LCRG1.


Pegylated liposomal formulation of doxorubicin overcomes drug resistance in a genetically engineered mouse model of breast cancer.
Success of cancer treatment is often hampered by the emergence of multidrug resistance (MDR) mediated by P-glycoprotein (ABCB1/Pgp). Doxorubicin (DOX) is recognized by Pgp and therefore it can induce therapy resistance in breast cancer patients. In this study our aim was to evaluate the susceptibility of the pegylated liposomal formulation of doxorubicin (PLD/Doxil®/Caelyx®) to MDR. We show that cells selected to be resistant to DOX are cross-resistant to PLD and PLD is also ineffective in an allograft model of doxorubicin-resistant mouse B-cell leukemia. In contrast, PLD was far more efficient than DOX as reflected by a significant increase of both relapse-free and overall survival of Brca1-/-;p53-/- mammary tumor bearing mice. Increased survival could be explained by the delayed onset of drug resistance. Consistent with the higher Pgp levels needed to confer resistance, PLD administration was able to overcome doxorubicin insensitivity of the mouse mammary tumors. Our results indicate that the favorable pharmacokinetics achieved with PLD can effectively overcome Pgp-mediated resistance, suggesting that PLD therapy could be a promising strategy for the treatment of therapy-resistant breast cancer patients.


Determination of drug concentrations in plasma by a highly automated, generic and flexible protein precipitation and liquid chromatography/tandem mass spectrometry method applicable to the drug discovery environment.
This report presents a highly automated procedure for the determination of drug concentrations in plasma samples. The method is generic, in that it has been applied without adaptation to many different drug candidate molecules, but is also flexible, in that variations in the nature and number of samples to be analyzed can be readily accommodated. The method includes preparation of dilutions of analyte stock solutions, spiking these into control plasma to generate analytical standards, and preparation of samples suitable for analysis by high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) by precipitation of plasma proteins with acetonitrile, centrifugation, and dilution of the supernatants with HPLC buffer. All of these steps, apart from centrifugation, are performed without manual intervention on an automated liquid-handling workstation using 96-well plates. Analysis is by HPLC/MS/MS, using a generic HPLC gradient. Commercially available software was used for optimization of parameters for analysis by HPLC/MS/MS, integration of chromatographic peaks, and quantification of drug concentrations. The use of this methodology in our laboratory has greatly facilitated the analysis of small sample sets for a large number of analytes, a situation regularly encountered in an early drug discovery environment.


Antimicrobial resistance in Europe and its potential impact on empirical therapy.
The problem of microbial drug resistance is a major public health concern, due to its global dimension and alarming magnitude, although the epidemiology of resistance can exhibit remarkable geographical variability and rapid temporal evolution. The major resistance issues overall are those related to methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), Enterobacteriaceae producing extended-spectrum beta-lactamases, and multidrug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii. Europe is not free from any of these issues, although their impact may be significantly different in different countries. MRSA rates are high in several European countries, but seem to have levelled off in some settings. Diffusion of VRE is still irregular. The most alarming resistance trends are those observed for Enterobacteriaceae and the Gram-negative non-fermenters, with a generalized increase in rates of resistance to the most important anti-Gram-negative agents (beta-lactams and fluoroquinolones) and the circulation of strains showing multidrug resistance phenotypes.


Apigenin Attenuates Adriamycin-Induced Cardiomyocyte Apoptosis via the PI3K/AKT/mTOR Pathway.
Treatment with Adriamycin (ADR) is one of the major causes of chemotherapy-induced cardiotoxicity and therefore is the principal limiting factor in the effectiveness of chemotherapy for cancer patients. Apigenin (API) has been shown to play a cardioprotective role. The present study examined the effect of API on ADR-induced cardiotoxicity in mice. Sixty male Kunming mice were randomly divided into 4 groups: a control group, ADR model group, low-dose API treatment group (125 mg·kg-1), and high-dose API treatment group (250 mg·kg-1). Blood samples were taken to evaluate a spectrum of myocardial enzymes. Cardiomyocyte apoptosis was measured using a TUNEL assay, and cardiomyocyte autophagy was observed using electron microscopy. Moreover, apoptosis-related proteins, such as Bax and Bcl-2, autophagy-related proteins, including Beclin1 and LC3B, and PI3K/AKT/mTOR pathway-related proteins were examined with western blot. Our results demonstrate that ADR caused an increase in the serum levels of cardiac injury markers and enhanced cardiomyocyte apoptosis and autophagy. API administration prevented the effects associated with ADR-induced cardiotoxicity in mice and inhibited ADR-induced apoptosis and autophagy. API also promoted PI3K/AKT/mTOR pathway activity in ADR-treated mice. In conclusion, API may have a protective effect against ADR-induced cardiotoxicity by inhibiting apoptosis and autophagy via activation of the PI3K/AKT/mTOR pathway.


Talonavicular coalition. Objective gait analysis.
Talonavicular coalitions, a rarely reported fusion between the talus and navicular, are often an incidental radiographic finding that may be asymptomatic or associated with peroneal spasm. The authors present a review of literature and case report based on clinical evaluation and instrumented gait analysis. Specifically, a patient presenting with a bilateral talonavicular coalition was objectively evaluated with kinetic, kinematic, muscle dynamometry, and pedobarographic testing to understand the biomechanical limitations related to this pathology. An excessive passive component of ankle torque, a high first metatarsophangeal joint plantar pressure, and a diminished time in the midstance portion of stance phase were measured and compared to those of healthy individuals.


The silkworm gustatory receptor BmGr63 is dedicated to the detection of isoquercetin in mulberry.
Gustatory systems in phytophagous insects are used to perceive feeding stimulants and deterrents, and are involved in insect decisions to feed on particular plants. During the process, gustatory receptors (Grs) can recognize diverse phytochemicals and provide a molecular basis for taste perception. The silkworm, as a representative Lepidoptera species, has developed a strong feeding preference for mulberry leaves. The mulberry-derived flavonoid glycoside, isoquercetin, is required to induce feeding behaviours. However, the corresponding Grs for isoquercetin and underlying molecular mechanisms remain unclear. In this study, we used molecular methods, voltage clamp recordings and feeding assays to identify silkworm BmGr63, which was tuned to isoquercetin. The use of qRT-PCR confirmed that BmGr63 was highly expressed in the mouthpart of fourth and fifth instar larvae. Functional analysis showed that oocytes expressing BmGr63 from the 'bitter' clade responded to mulberry extracts. Among 20 test chemicals, BmGr63 specifically recognized isoquercetin. The preference for isoquercetin was not observed in BmGr63 knock-down groups. The tuning between BmGr63 and isoquercetin has been demonstrated, which is meaningful to explain the silkworm-mulberry feeding mechanism from molecular levels and thus provides evidence for further feeding relationship studies between phytophagous insects and host plants.


Expression and regulation of integrin receptors in human trophoblast cells: role of estradiol and cytokines.
Embryo implantation and placentation are dynamic cellular events that require not only synchrony between the maternal environment and the embryo, but also complex cell-cell communication amongst the implanting blastocyst and the receptive endometrium through integrins, a large family of proteins involved in the attachment, migration, invasion and control of cellular functions. Integrins display dynamic temporal and spatial patterns of expression by the trophoblast cells during early pregnancy in humans. However, the precise mechanism of embryo implantation and the modulation of the integrin receptors during blastocyst attachment and further implantation remain elusive in the humans. The present study elucidates the expression and hormonal modulation of fibronectin, vitronectin and laminin integrin receptors by estradiol and IL-1alpha in human trophoblast cells. Human first trimester trophoblast cells showed the induction of the classical estrogen receptor (ER)-alpha by its own ligand, estradiol. Treatment with either estradiol or IL-1alpha induced the expressions of alpha4, alpha5, alpha6 and alpha(v) integrin receptor subunits at both the mRNA and protein levels, while expression of beta1 remained unaltered. Furthermore, estradiol upregulated the expression of IL-1alpha, thereby suggesting the possibility that estrogen may either directly or via the proinflammatory cytokine induces the expression of the cell surface integrin receptors. The findings delineate the role of hormones and the cytokines in modulating the adhesiveness and attachment of the trophoblast cells. This may reflect the in vivo scenario where the implanting embryo is surrounded by a hormone-cytokine rich uterine microenvironment that may precisely regulate the expression of integrins and thereby facilitate implantation.


Inhibition of radiogenic and chemically induced transformation in C3H/10T-1/2 cells by a polyprenoic acid (E-5166).
Using C3H/10T-1/2 mouse fibroblasts, we tested whether polyprenoic acid (E-5166) inhibits radiogenic and chemically induced transformation in vitro. Our results show that E-5166 markedly inhibits transformation by X rays and benzo[a]pyrene in a dose-related manner. Maximum inhibition was observed when cells were pretreated with E-5166 prior to carcinogen exposure, and lesser inhibition when E-5166 treatment followed carcinogen exposure. These results indicate that E-5166 can serve as a radio-protective and chemopreventive agent with anticarcinogenic potential.


[Morphological and biochemical studies of collagen in prostatic hypertrophy (author's transl)].
In an attempt to clarify the differentiation of prostate collagen under the comparison with histological pictures, 46 human prostatic hypertrophy specimens (from 50 to 79 years old) and normal specimens (from 27 gestational weeks to 80 years old) were examined by these techniques; the determination of hydroxyproline, polyacrylamide gel electrophoresis, CM-cellulose chromatography, amino acid analysis, and electron-microscopical studies on Segment-Long-Spacing collagen. The prostatic hypertrophy specimens were morphologically divided into stromal type (13 cases), glandular type (11 cases), and mixed type (22 cases). The results were as follows: 1) The prostate collagen was composed of Type I and Type III collagens in both hypertrophic and normal group. 3) In normal group, the average value of the lysyl hydroxylation of collagen in fetus and newborn prostate was 29.8 +/- 1.1% (mean +/- S.E.), but it showed conspicuous decrease in childhood and indicated about 18% by 20 years old. 4) The average value in hypertrophic group was 28.6 +/- 0.7%, while that in the age normal people over 50 years old was 22.8 +/- 2.3%, These findings led to the conclusion that an early stage collagen increases in prostatic hypertrophy, therefore it is presumed that there is a defect in process of collagen maturation.


Spontaneous vegetative endocarditis due to Enterococcus faecalis in a rottweiler puppy.
A 5-month-old female Rottweiler was referred because of a 5-week diarrhea and a sudden onset of a cardiac murmur auscultated by its veterinarian. Definitive diagnosis of bacterial endocarditis was based on ultrasonographic visualization of vegetative cardiac lesions and positive cultures of Enterococcus faecalis in blood and urine. Complicating findings were suppurative nephritis and renal infarction. Despite intensive supportive care, the endocarditis and clinical condition deteriorated and the dog had to be euthanized.


Consolidated bioprocessing of AFEX-pretreated corn stover to ethanol and hydrogen in a microbial electrolysis cell.
The consolidated bioprocessing (CBP) of corn stover pretreated via ammonia fiber expansion (AFEX-CS) into ethanol was investigated in a microbial electrolysis cell (MEC) driven by the exoelectrogen Geobacter sulfurreducens and the CBP bacterium Cellulomonas uda. C. uda was identified in a screening for its ethanologenic potential from AFEX-CS and for producing electron donors for G. sulfurreducens fermentatively. C. uda produced ethanol from AFEX-CS in MECs inoculated simultaneously or sequentially, with the concomitant conversion of the fermentation byproducts into electricity by G. sulfurreducens. The fermentation and electrical conversion efficiencies were high, but much of the AFEX-CS remained unhydrolyzed as nitrogen availability limited the growth of the CBP partner. Nitrogen supplementation stimulated the growth of C. uda, AFEX-CS hydrolysis and ethanologenesis. As a result, the synergistic activities of the CBP and exoelectrogen catalysts resulted in substantial energy recoveries from ethanologenesis alone (ca. 56%). The cogeneration of cathodic H(2) in the MEC further increased the energy recoveries to ca. 73%. This and the potential to optimize the activities of the microbial catalysts via culturing approaches and genetic engineering or adaptive evolution, make this platform attractive for the processing of agricultural wastes.


Agmatine Alleviates Epileptic Seizures and Hippocampal Neuronal Damage by Inhibiting Gasdermin D-Mediated Pyroptosis.
Background: Epilepsy is a common neurological disease, and neuroinflammation is one of the main contributors to epileptogenesis. Pyroptosis is a type of pro-inflammatory cell death that is related to epilepsy. Agmatine, has anti-inflammatory properties and exerts neuroprotective effects against seizures. Our study investigated the effect of agmatine on the core pyroptosis protein GSDMD in the context of epilepsy. Methods: A chronic epilepsy model and BV2 microglial cellular inflammation model were established by pentylenetetrazole (PTZ)-induced kindling or lipopolysaccharide (LPS) stimulation. H&E and Nissl staining were used to evaluate hippocampal neuronal damage. The expression of pyroptosis and inflammasome factors was examined by western blotting, quantitative real-time PCR, immunofluorescence and enzyme-linked immunosorbent assay (ELISA). Results: Agmatine disrupted the kindling acquisition process, which decreased seizure scores and the incidence of full kindling and blocked hippocampal neuronal damage. In addition, agmatine increased BV2 microglial cell survival in vitro and alleviated seizures in vivo by suppressing the levels of PTZ-induced pyroptosis. Finally, the expression of TLR4, MYD88, phospho-IκBα, phospho-NF-κB and the NLRP3 inflammasome was significantly upregulated in LPS-induced BV2 microglial cells, while agmatine suppressed the expression of these proteins. Conclusions: Our results indicate that agmatine affects epileptogenesis and exerts neuroprotective effects by inhibiting neuroinflammation, GSDMD activation, and pyroptosis. The inhibitory effect of agmatine on pyroptosis was mediated by the suppression of the TLR4/MYD88/NF-κB/NLRP3 inflammasome pathway. Therefore, agmatine may be a potential treatment option for epilepsy.


Multicomponent Reactions with Cyclic Tertiary Amines Enabled by Facile C-N Bond Cleavage.
A novel and catalyst-free multicomponent reaction with cyclic tertiary amines, electron-deficient aryl halides or heteroaromatic halides, and Na2 S enabled by facile C-N bond cleavage of the cyclic tertiary amines was developed. This direct and operationally simple method can be applied with a wide range of functional groups and provides an efficient and rapid approach to potentially drug-like products containing amine, azaarene, thioether, or phenol ether functionalities in good to excellent yields. The utility of this method was demonstrated by the rapid synthesis of the analgesic ruzadolane.


Development of a rapid and efficient method for non-lethal DNA sampling and genotyping in scallops.
Non-lethal DNA sampling has long appealed to researchers studying population and conservation genetics, as it does not necessitate removing individuals permanently from their natural environment or destroying valuable samples. However, such an approach has not yet been well established in bivalves. In this study, we demonstrate that the gill represents a good source of tissue for non-lethal sampling in scallops. Removal of a few gill filaments caused no noticeable behavioral abnormalities or increased mortality rates in Zhikong scallop (Chlamys farreri) during a three-month period of observation. To facilitate rapid gill-based DNA extraction, six methods (MA-MF) were designed and evaluated, each requiring less than one hour of processing time. The optimal method was identified as MF, in terms of maintaining DNA integrity and genotyping accuracy. Further optimization of MF method by orthogonal experimental design suggested that the utilization of gills could be limited to 2 mg of sample, which is sufficient for performing up to 20,000 PCR reactions. We also demonstrate the excellent cross-species utility of MF in two additional scallop species, Yesso scallop (Patinopecten yessoensis) and bay scallop (Argopecten irradians). Taken together, our study provides a rapid and efficient approach for applying non-lethal DNA sampling in bivalve species, which would serve as a valuable tool for maintaining bivalve populations and conservation genetics, as well as in breeding studies.


A small heat-shock protein (Hsp20) regulated by RpoS is essential for cyst desiccation resistance in Azotobacter vinelandii.
In Azotobacter vinelandii, a cyst-forming bacterium, the alternative sigma factor RpoS is essential to the formation of cysts resistant to desiccation and to synthesis of the cyst-specific lipids, alkylresorcinols. In this study, we carried out a proteome analysis of vegetative cells and cysts of A. vinelandii strain AEIV and its rpoS mutant derivative AErpoS. This analysis allowed us to identify a small heat-shock protein, Hsp20, as one of the most abundant proteins of cysts regulated by RpoS. Inactivation of hsp20 did not affect the synthesis of alkylresorcinols or the formation of cysts with WT morphology; however, the cysts formed by the hsp20 mutant strain were unable to resist desiccation. We also demonstrated that expression of hsp20 from an RpoS-independent promoter in the AErpoS mutant strain is not enough to restore the phenotype of resistance to desiccation. These results indicate that Hsp20 is essential for the resistance to desiccation of A. vinelandii cysts, probably by preventing the aggregation of proteins caused by the lack of water. To our knowledge, this is the first report of a small heat-shock protein that is essential for desiccation resistance in bacteria.


Understanding the implementation and adoption of a technological intervention to improve medication safety in primary care: a realist evaluation.
Monitoring for potentially hazardous prescribing is increasingly important to improve medication safety. Healthcare information technology can be used to achieve this aim, for example by providing access to prescribing data through surveillance of patients' electronic health records. The aim of our study was to examine the implementation and adoption of an electronic medicines optimisation system that was intended to facilitate clinical audit in primary care by identifying patients at risk of an adverse drug event. We adopted a sociotechnical approach that focuses on how complex social, organisational and institutional factors may impact upon the use of technology within work settings. We undertook a qualitative realist evaluation of the use of an electronic medicines optimisation system in one Clinical Commissioning Group in England. Five semi-structured interviews, four focus groups and one observation were conducted with a range of stakeholders. Consistent with a realist evaluation methodology, the analysis focused on exploring the links between context, mechanism and outcome to explain the ways the intervention might work, for whom and in what circumstances. Using the electronic medicines optimisation system could lead to a number of improved patient safety outcomes including pre-emptively reviewing patients at risk of adverse drug events. The effective use of the system depended upon engagement with the system, the flow of information between different health professionals centrally placed at the Clinical Commissioning Group and those locally placed at individual general practices, and upon variably adapting work practices to facilitate the use of the system. The use of the system was undermined by perceptions of ownership, lack of access, and lack of knowledge and awareness. The use of an electronic medicines optimisation system may improve medication safety in primary care settings by identifying those patients at risk of an adverse drug event. To fully realise the potential benefits for medication safety there needs to be better utilisation across primary care and with a wider range of stakeholders. Engaging with all potential stakeholders and users prior to implementation of such systems might allay perceptions that the system is owned centrally and increase knowledge of the potential benefits.