Development of a bi-layered cryogenic electrospun polylactic acid scaffold to study calcific aortic valve disease in a 3D co-culture model.
Calcified aortic valve disease (CAVD) is the most prevalent valve disease in the elderly. Targeted pharmacological therapies are limited since the underlying mechanisms of CAVD are not well understood. Appropriate 3D in vitro models could potentially improve our knowledge of the disease. Here, we developed a 3D in vitro aortic heart valve model that resembles the morphology of the valvular extracellular matrix and mimics the mechanical and physiological behavior of the native aortic valve fibrosa and spongiosa. We employed cryogenic electrospinning to engineer a bi-layered cryogenic electrospun scaffold (BCES) with defined morphologies that allowed valvular endothelial cell (VEC) adherence and valvular interstitial cell (VIC) ingrowth into the scaffold. Using a self-designed cell culture insert allowed us to establish the valvular co-culture simultaneously by seeding VICs on one side and VECs on the other side of the electrospun scaffold. Proof-of-principle calcification studies were successfully performed using an established osteogenic culture protocol and the here designed 3D in vitro aortic heart valve model. STATEMENT OF SIGNIFICANCE: Three-dimensional (3D) electrospun scaffolds are widely used for soft tissue engineering since they mimic the morphology of the native extracellular matrix. Several studies have shown that cells behave more naturally on 3D materials than on the commonly used stiff two-dimensional (2D) cell culture substrates, which have no biological properties. As appropriate 3D models for the study of aortic valve diseases are limited, we developed a novel bi-layered 3D in vitro test system by using the versatile technique of cryogenic electrospinning in combination with the influence of different solvents to mimic the morphology, mechanical, and cellular distribution of a native aortic heart valve leaflet. This 3D in vitro model can be used to study valve biology and heart valve-impacting diseases such as calcification to elucidate therapeutic targets.


MicroRNAs miR-186 and miR-150 down-regulate expression of the pro-apoptotic purinergic P2X7 receptor by activation of instability sites at the 3'-untranslated region of the gene that decrease steady-state levels of the transcript.
The P2X7 receptor regulates cell growth through mediation of apoptosis. P2X7 levels are lower in cancer epithelial cells than in normal cells, and previous studies showed that expression of P2X7 was regulated post-transcriptionally. The objective of the study was to understand regulation of P2X7 mRNA stability. Overexpression of a reporter containing the full-length human P2X7 3'-untranslated region (3'-UTR) or reporters containing parts of the 3'-UTR-P2X7 were associated with increased abundance of the construct in normal cells and decreased abundance in cancer epithelial cells. Sequences within the 3'-UTR-P2X7, which are putative target sites for the microRNAs, miR-186 (middle segment) and miR-150 (distal segment), decreased the abundance of the P2X7 transcript. Overexpression in cancer cells of mutated miR-186 and miR-150 target sites was associated with lower levels of the reporter genes. In normal cells overexpression of the mutated miR-186 target site was associated with marked increased concentration, but overexpression of the miR-150 target site reporters, wild-type and mutant, did not change over time. Levels of miR-186 and miR-150 were higher in cancer than in normal cells, and treatment with miR-186 and miR-150 inhibitors increased P2X7 mRNA. In human embryonic kidney-293 cells heterologously expressing the full-length 3'-UTR-P2X7 luciferase reporter, miR-186 and miR-150 inhibitors increased luciferase activity, whereas miR-186 and miR-150 mimics decreased luciferase activity after actinomycin D treatment. These data suggest that increased expression of miR-186 and miR-150 in cancer epithelial cells decreases P2X7 mRNA by activation of miR-186 and miR-150 instability target sites located at the 3'-UTR-P2X7.


[Prevention of bleeding during tubeless percutaneous nephrolithotomy].
Nephrostomy tube placed after percutaneous removal of kidney stones can cause pain and discomfort in the early postoperative period [1, 2]. A tubeless percutaneous nephrolithotomy (PCNL) is considered as alternative approach. However, the most serious complication after tubeless procedure is a development of active uncontrolled bleeding from percutaneous tract in the early postoperative period. To improve the results of surgical treatment of kidney stones by evaluation of efficiency of using hemostatic matrix during tubeless PCNL. A total of 113 patients with large kidney stones undergone to the tubeless PCNL for the past 9 years in our clinic. The external or internal stent was put for the drainage of upper urinary tract for 2 and 14 days, respectively. All patients were divided into 2 groups, depending on severity of bleeding (group 1 - no bleeding vs. group 2 - mild bleeding). Each group was further divided into two subgroups based on the technique and type of drainage of upper urinary tract at the end of the surgery. In the main group (n=74) the hemostatic matrix based on lyophilized thrombin 2000 IU was injected in percutaneous tract at the end of the surgery. The control group included 39 patients who underwent mini-PCNL without using hemostatic matrix. The blood loss, intensity of urine leakage from percutaneous tract (duration, visual analog scale) and degree of fluid extravasation in pararenal fat estimated by US were evaluated. The mean operative time was 47.5+/-3.6 min (52, 58, 38 and 49 min in subgroup I, II, III and control group, respectively). Hemoglobin drop was 9.3+/-4.1, 12.1+/-7.5, 14.6+/-11.2 and 10.6+/-5.9 in subgroups I, II, III and in the control group, respectively, while duration of hematuria was 16.1, 20.3, 28.5 and 22.9 hours, respectively. Fluid extravasation in pararenal space was found in 1 out 7 patients in subgroup III and control group, respectively (p<0.05). The dilatation of collection system (after double-J) persisted in 18.1%, 20% 22.2% and 13.3% cases in subgroup I, II, III and control group, respectively. There were no significant differences in other evaluated parameters. In 1 case in the control group there was severe bleeding which required selective embolization of the renal artery. The use of hemostatic matrix is an additional important measure which allows to prevent active bleeding from the percutaneous tract in early postoperative period. Moreover, the hemostatic matrix contributes to the sealing of tract and reduces the risk of fluid extravasation in pararenal space after mini-PCNL.


Effects of several antibiotics on the neuromuscular junction: Part II.
The effects of various kinds of antibiotics including tetracycline (TC), chloramphenicol (CP), sodium cephalothin (CET), sodium cefazolin (CEZ), colistin sulfate (CL), colistin sodium methanesulfonate (CL-M), bacitracin (BC), gramicidin HCl (GR), rifampicin sulfate (RFP) and lincomycin (LCM), on the neuromuscular junction (NMJ) were studied by in vitro and in vivo experiments. In in vitro experiments, CL and LCM exhibited a blocking effect on the NMJ in rat phrenic nerve diaphragm preparations, and GR caused a marked increase in muscle contraction. This effect was not affected by administration of eserine or CaCl2. In in vitro experiments with frog sciatica nerve and musculus sartorius preparations, CL and GR induced the appearance of endplate potentials, suggesting blockade of the NMJ. No blocking effect of other antibiotics was observed. In in vitro experiments with the preparations from Rana catesbiana frogs, TC and LCM induced a decrease in the frequency of miniature endplate potentials. In in vivo experiments with rabbit musculus tibialis anterior preparations, CL, TC and LCM exerted a blocking effect soon after administration, but GR and RFP had a late blocking effect. CL, GR, BC and RFP were found not to compete with eserine or CaCl2 in terms of the blocking effect on the NMJ. From the fact that TC did not compete with eserine but did compete with CaCl2 and with KCl as blockers at the NMJ, this blocking effect of TC seems to be due to inhibition of release of acetylcholine (ACh). The fact that LCM competes with eserine indicates that this antibiotic has the same type of action as curare on ACh receptors of the NMJ.(ABSTRACT TRUNCATED AT 250 WORDS)


Construction of stable MoxSy/CeO2 heterostructures for the electrocatalytic hydrogen evolution reaction.
The unique structures of polynuclear MoxSy clusters make it possible to maximize the number of their active sites and for them to be good candidates for HER catalysts. An appropriate support is highly necessary not only to avoid the desorption of MoxSy clusters in a working environment, but also to improve their HER activity. Our work here shows that the CeO2 support could provide strong support for interaction with various MoxSy clusters and the formed MoxSy/CeO2 hetero-structures also have modest ΔGH* for the HER. The electronic features of MoxSy clusters are regulated by the CeO2 support, which leads to charge redistribution on edge atoms and plays a key role in H adsorption. Our studies provide instructive predictions on efficient candidates of molybdenum-sulfur based catalysts for the HER.


Clozapine and Fever: A Case of Continued Therapy With Clozapine.
Clozapine is a major atypical antipsychotic drug used in treatment-resistant schizophrenia (Patel and Allin. Ther Adv Psychopharmacol 2011;1:25-29). It interferes with dopamine binding to D1, D2, D3, and D5 receptors but has high affinity to D4. It also has an anticholinergic effect and antagonizes α-adrenergic, histaminergic, and serotoninergic receptors (Oerther and Ahlenius. J Pharmacol Exp Ther 2000;292:731-736). Clozapine has proved effective in treating positive and negative symptoms in patients with refractory schizophrenia, thus accounting for its frequent use. Despite its effectiveness, this drug is not without its adverse effects. The most well known is agranulocytosis. There are, however, many others, such as myocarditis, aspiration pneumonia, ileus, fever, hyperglycemia, hyperlipidemia, hypertriglycemia, tachycardia, and weight gain, among others (Bruijnzeel et al. Asian J Psychiatr 2014;11:3-7). Fever induced by clozapine is a common phenomenon (Lowe et al. Ann Pharmacother 2007;41:1700-1704), which usually occurs in the first 4 weeks of treatment, and its prevalence oscillates from 0.5% and 55%, depending on the study (Jeong et al. Schizophr Res 2002;56:191-193; Young et al. Schizophr Bull 1998;24:381-390). The fever lasts for 2.5 days on average, and unless the treatment is discontinued, it generally abates between day 8 and 16 of treatment (Kohen et al. Ann Pharmacother 2009;43:143-146). There are several different theories about the physiopathological mechanism; it could be a variation of malignant neuroleptic syndrome, an infection secondary to neutropenia, and allergic reaction or the emergence of the immunomodulating effect of clozapine. Some case reports in the bibliography have shown that patients in treatment with clozapine can develop a mild leukocytosis, but the presence of other concurrent symptoms, which indicate infection, is not common (Tham and Dickson. J Clin Psychiatry 2002;63:880-884). The theory of an allergic reaction is unsupported because of the fact that the fever does not recur after reintroducing clozapine. So we question, "What would be the attitudes to follow when we find clozapine-induced fever (Nielsen et al. J Clin Psychiatry 2013;74:603-613)?" The management of patients with clozapine-induced fever should include a complete blood picture, liver and renal function tests, a creatine kinase test urine culture, and a chest x-ray. A nasopharyngeal aspirate can also be useful to exclude infection (Pui-yin Chung et al. Can J Psychiatry 2008;53:857-862). On the other hand, some drugs have been suggested for treatment of fever induced by clozapine. The use of acetaminophen, in the treatment of the fever induced by clozapine, is supported by many studies (Jeong et al. Schizophr Res 2002;56:191-193). In one of these, clozapine was suspended and restarted successfully after 1 week. However, in some studies, such as the case report of Tremeau et al (Clin Neuropharmacol 1997;20:168-170), clozapine was reduced instead of discontinued. In other studies, the recommendation is continuating clozapine treatment (Martin and Williams. J Psychiatry Neurosci 2013;38:E9-E10).


Phenytoin: effective but insidious therapy for epilepsy in people with intellectual disability.
Phenytoin (5,5-diphenylhydantoin), which has been in use for 60 years, is still an important antiepileptic drug. Its primary mechanism of action is modulation of the sustained repetitive firing of neurones by direct inhibition and blockage of voltage-gated sodium channels in the neuronal cell membrane, and by delay of cellular reactivation. The plasma protein binding of phenytoin is normally between 90% and 95%. The drug is rapidly distributed from the blood to the tissues and is almost completely metabolized in the liver. The plasma phenytoin concentration normally reaches the steady-state level within 1-2 weeks. The half-life of phenytoin is less than 20 h in low doses, but is prolonged in high doses, newborn infants and elderly people. The half-life is shortened when phenytoin is given concomitantly with an enzyme-inducing drug, such as phenobarbital or carbamazepine. Phenytoin is effective for treating generalized tonic-clonic seizures, partial seizures with or without generalization, and convulsive status epilepticus. Over the years, many new, and even serious, adverse effects of phenytoin have been recognized. Phenytoin encephalopathy, manifesting as cognitive impairment and a cerebellar syndrome, is an important adverse neurological effect, the development of which depends on the saturation kinetics of phenytoin, individual differences in phenytoin metabolism, an inhibitory effect of certain drugs on phenytoin metabolism, or the ability of certain drugs to displace phenytoin from plasma proteins, leading to an increase in the plasma level of unbound phenytoin. Because of its potentially adverse effects, phenytoin is not recommended as the first choice for treating epileptic seizures, except as a co-drug for managing convulsive status epilepticus. In patients with epilepsy who also have intellectual disability, and are susceptible to balance disturbances and cognitive dysfunction, it is wise to replace phenytoin with another drug, such as carbamazepine or oxcarbazepine. The long-term use of phenytoin is not recommended for patients with loss of locomotion, marked cognitive impairment, or symptoms and signs of cerebellar disease. The prevention of phenytoin intoxication, with the subsequent development of phenytoin-induced encephalopathy, depends on careful observation of the patients and frequent monitoring of plasma levels of phenytoin and other concomitantly administered antiepileptic drugs.


Role of kappa-opioid and mu-opioid receptors in pruritus: Peripheral and central itch circuits.
Modern genetic approaches in animal models have unveiled novel itch-specific neural pathways, emboldening a paradigm in which drugs can be developed to selectively and potently target itch in a variety of chronic pruritic conditions. In recent years, kappa-opioid receptors (KORs) and mu-opioid receptors (MORs) have been implicated in both the suppression and promotion of itch, respectively, by acting on both the peripheral and central nervous systems. The precise mechanisms by which agents that modulate these pathways alleviate itch remains an active area of investigation. Notwithstanding this, a number of agents have demonstrated efficacy in clinical trials that influence both KOR and MOR signalling. Herein, we summarize a number of opioid receptor modulators in development and their promising efficacy across a number of chronic pruritic conditions, such as atopic dermatitis, uremic pruritus and beyond.


Synthetic inhibitors of endopeptidase EC 18.104.22.168: potency and stability in vitro and in vivo.
1. The role of the metalloendopeptidase EC 184.108.40.206 (EP 24.15) in peptide metabolism in vivo is unknown, in part reflecting the lack of a stable enzyme inhibitor. The most commonly used inhibitor, N-[1-(R,S)-carboxy-3-phenylpropyl]-Ala-Ala-Tyr-p-aminobenzoate (cFP-AAY-pAB, Ki = 16 nM), although selective in vitro, is rapidly degraded in the circulation to cFP-Ala-Ala, an angiotensin converting enzyme (ACE) inhibitor. This metabolite is thought to be generated by neutral endopeptidase (NEP; EC 184.108.40.206), as the Ala-Tyr bond of cFP-AAY-pAB is cleaved by NEP in vitro. In the present study, we have examined the role of NEP in the metabolism of cFP-AAY-pAB in vivo, and have tested a series of inhibitor analogues, substituted at the second alanine, for both potency and stability relative to the parent compound. 2. Analogues were screened for inhibition of fluorescent substrate cleavage by recombinant rat testes EP 24.15. D-Ala or Asp substitution abolished inhibitory activity, while Val-, Ser- and Leu-substituted analogues retained activity, albeit at a reduced potency. A relative potency order of Ala (1) > Val (0.3) > Ser (0.16) > Leu (0.06) was observed. Resistance to cleavage by NEP was assessed by incubation of the analogues with rabbit kidney membranes. The parent compound was readily degraded, but the analogues were twice (Ser) and greater than 10 fold (Leu and Val) more resistant to cleavage. 3. Metabolism of cFP-AAY-pAB and the Val-substituted analogue was also examined in conscious rabbits. A bolus injection of cFP-AAY-pAB (5 mg kg-1, i.v.) significantly reduced the blood pressure response to angiotensin I, indicating ACE inhibition. Pretreatment with NEP inhibitors, SCH 39370 or phosphoramidon, slowed the loss of cFP-AAY-pAB from the plasma, but did not prevent inhibition of ACE. Injection of 1 mg kg-1 inhibitor resulted in plasma concentrations at 10 s of 23.5 microM (cFP-AAY-pAB) and 18.0 microM (cFP-AVY-pAB), which fell 100 fold over 5 min. Co-injection of 125I-labelled inhibitor revealed that 80-85% of the radioactivity had disappeared from the circulation within 5 min, and h.p.l.c. analysis demonstrated that only 25-30% of the radiolabel remained as intact inhibitor at this time. Both analogues were cleared from the circulation at the same rate, and both inhibitors blunted the pressor response to angiotensin I, indicative of ACE inhibition. 4. These results suggest that both NEP and other clearance/degradation mechanisms severely limit the usefulness of peptide-based inhibitors such as cFP-AAY-pAB. To examine further EP 24.15 function in vivo, more stable inhibitors, preferably non-peptide, must be developed, for which these peptide-based inhibitors may serve as useful molecular templates.


Command and compensation in a neuromodulatory decision network.
The neural circuits that mediate behavioral choices must not only weigh internal demands and environmental circumstances, but also select and implement specific actions, including associated visceral or neuroendocrine functions. Coordinating these multiple processes suggests considerable complexity. As a consequence, even circuits that support simple behavioral decisions remain poorly understood. Here we show that the environmentally sensitive wing expansion decision of adult fruit flies is coordinated by a single pair of neuromodulatory neurons with command-like function. Targeted suppression of these neurons using the Split Gal4 system abrogates the fly's ability to expand its wings in the face of environmental challenges, while stimulating them forces expansion by coordinately activating both motor and neuroendocrine outputs. The arbitration and implementation of the wing expansion decision by this neuronal pair may illustrate a general strategy by which neuromodulatory neurons orchestrate behavior. Interestingly, the decision network exhibits a plasticity that is unmasked under conducive environmental conditions in flies lacking the function of the command-like neuromodulatory neurons. Such flies can often expand their wings using a motor program distinct from that of wild-type animals and controls. This compensatory program may be the vestige of an ancestral, environmentally insensitive program used for wing expansion that existed before the evolution of the environmentally adaptive program currently used by Drosophila and other cyclorrhaphan flies.


The involvement of the opioid system in human obesity: a study in normal weight relatives of obese people.
The involvement of the opioid system in human obesity has been demonstrated, but whether the abnormalities in the endorphinergic system play a primary role in overfeeding and weight gain or represent a simple biochemical feature is still unclear. The objectives of this study were to investigate the effects of both physiological and pharmacological plasma beta-endorphin levels on some metabolic and hormonal parameters in a normal weight, but prone to obesity, young population consisting of first degree relatives of obese subjects and in body mass index-, sex, and age- matched control subjects without a family history of obesity. Each subject underwent a 1-h infusion of synthetic human beta-endorphin at a constant rate of 4.5 ng/kg.min (low rate), then after a 1-week interval, at a rate of 500 micrograms/h (high rate). Under basal conditions, there was no significant difference in plasma glucose and pancreatic hormones (insulin, C peptide, and glucagon) between the two groups, except for plasma beta-endorphin levels, which were significantly (P < 0.01) higher in relatives of obese individuals. The low rate of beta-endorphin infusion induced physiological elevations of plasma opioid levels in both groups; no significant change in plasma glucose and pancreatic products in control subjects; and a significant (at least P < 0.05) rise in plasma insulin, C peptide, and glucagon concentrations in relatives of the obese. The high rate of beta-endorphin infusion produced pharmacological elevations of opioid plasma levels in both groups; significant (at least P < 0.05) increments in plasma glucose and glucagon levels and no appreciable modification of plasma insulin and C peptide levels in the control group; and a significant (at least P < 0.05) positive response of plasma glucose, insulin, C peptide, and glucagon levels in relatives of obese subjects. These findings suggest that 1) opioid peptides at least in part play a primary, rather than secondary, role in some metabolic events of obesity; and 2) both physiological and pharmacological plasma levels of beta-endorphin are able to provoke marked islet hormone release in the early phase of human obesity.


A dose-finding approach based on shrunken predictive probability for combinations of two agents in phase I trials.
We develop a novel dose-finding method for two-agent combination phase I trials on the basis of the shrunken predictive probability of toxicity. In this method, a shrinkage logistic regression model that allows distinct shrinkage multipliers for the coefficients of the main effects of two agents and their interaction on the probability of toxicity constructs the toxicity outcome. We also propose dose-escalation/de-escalation decision rules on the basis of the shrunken predictive probability of toxicity. Simulation studies under various patterns of monotonic dose-response relationships for combinations of two agents demonstrated that the proposed method performed no worse than the existing two dose-finding methods we selected.


Identification of New Players in Hepatocarcinogenesis: Limits and Opportunities of Using Tissue Microarray (TMA).
Liver tumours are among the leading causes of cancer-related death worldwide and hepatocellular carcinoma (HCC) accounts for the vast majority of liver tumours. When detected at an early stage of disease, patients might still be eligible for surgical-based curative treatments. However, currently only small portion of HCC affected patients are diagnosed at an early stage. For late stage HCC no treatment option exists beside the multi-tyrosine kinase inhibitor Sorafenib. Thus new molecular targets and treatment options for HCC are urgently needed. Nevertheless, despite some improvements in diagnosis and patient management, the biology of liver tumour remains inadequately understood, mainly because these tumours have shown to harbour a highly complex genomic landscape. In addition, one major obstacle delaying the identification of new molecular targets in biomedical research is the necessity to validate them using a large collection of tissue specimens. Tissue microarray (TMA) technology allows the prompt molecular profiling of multiple tissue specimens and is therefore ideal to analyze presumptive candidate biomarkers in a fast an effective manner. The use of TMA has substantial benefits over standard techniques and represents a significant advancement in molecular pathology. For example, TMA technology reduces laboratory work, offers a high level of experimental uniformity and provides a judicious use of precious tissue. On the other hand, one potential limitation of using TMA is that the small cores sampled may not be representative of whole tumors. This issue is very critical in particularly heterogeneous cancers such as HCC. For liver focused studies, it is ideal to evaluate the staining patters of a determined marker over the structure of an entire acinus and to define staining in as many as possible anatomical regions. In this review we analyze the limits and opportunities offered by the usage of TMA technology in HCC research. In summary, TMA has revolutionized the histopathological analysis and will be of great help to further advance the knowledge in the field of hepatocarcinogenesis research.


Prevalence and risk factors for depression, anxiety and suicidal ideation in youth with persistent post-concussive symptoms (PPCS).
Objectives: To assess the prevalence and risk factors for emotional distress in youth with persistent post-concussive symptoms (PPCS) greater than one month.Methods: We used baseline data from an intervention study for youth with PPCS, utilizing Poisson regression to examine factors associated with exceeding clinical cut-points on measures of depression, anxiety, self-harm and suicidal ideation. Predictors included: age, sex, socioeconomic status, mental health history, duration of concussion symptoms, history of prior concussion, trauma history and sleep quality.Results: The sample included 200 youth with PPCS, (mean 14.7 SD 1.7 years, 82% white, 62% female). Forty percent reported clinically significant depressive symptoms, 25% anxiety, 14% thoughts of self-harm and 8% thoughts of suicide. History of depression was associated with 3-fold higher risk for thoughts of self-harm (95% CI:1.82-6.99) and 6-fold higher risk for suicidal ideation (95% CI:1.74-24.46). Better sleep quality was associated with lower risk for all outcomes. History of prior concussion and duration of PPCS were not significantly associated with any outcomes.Conclusions: Suicidal thoughts are common post-concussion, and history of depression is a strong risk factor. Tailored interventions may be needed to address mental health in this population.


Flu Vaccinations in Pharmacies-A Review of Pharmacists Fighting Pandemics and Infectious Diseases.
The phenomenon of population ageing observed over recent years involves growing healthcare needs and the limited staffing and financing of healthcare systems, and as such demands some functional changes in the healthcare model in many countries. This situation is particularly significant in the face of a pandemic, e.g., flu, and currently COVID-19.As well as social education, preventive vaccinations are the most effective method of fighting the infectious diseases posing a special threat to seniors. Despite this, the vaccination coverage level in most European countries is relatively low. This is largely due to patients having limited access to vaccinations. In some countries, implementing vaccinations in pharmacies and by authorized pharmacists has significantly improved vaccination coverage rates and herd immunity, while lowering the cost of treating infections and the resulting complications, as well as minimizing the phenomenon of inappropriate antibiotic therapies. This article presents the role of pharmacists in the prevention of infectious diseases, pointing out the measurable effects of engaging pharmacists in conducting preventive vaccinations, as well as analyzing the models of implementing and conducting vaccinations in pharmacies in selected countries, and depicting recommendations regarding vaccinations developed by international organizations. The presented data is used to suggest requirements for the implementation of preventive vaccinations in community pharmacies.


Hepatitis B Vaccination in Patients with Liver Cirrhosis Evaluated for Liver Transplantation - A Simple Intervention Ensures High Adherence.
BACKGROUND There is an unmet need to improve the HBV vaccination status in patients with chronic liver diseases. Primary care physicians and outpatient hepatologists often fail to vaccinate as recommended. Thus, new strategies to improve the HBV vaccination rate are required. MATERIAL AND METHODS This study was performed in a cohort of patients with chronic liver diseases evaluated for liver transplantation. Vaccination status was taken from the patients' vaccination cards. HBsAg-, anti-HBc-, and anti-HBs-negative individuals were vaccinated against HBV at hospital discharge, and subsequent outpatient completion of the standard vaccination protocol was recommended in detail in the discharge letter. At months 2 and 8, titer controls were performed, and completion of vaccination was evaluated. RESULTS We prospectively recruited 37 patients. At baseline, the vaccination rate against HBV was 24% (N=9/37), and 3/9 HBV vaccinated patients presented with an anti-HBs-titer >10 IU/L. Thus, N=34 were vaccinated with Engerix® or Twinrix®. We evaluated 26/34 patients at month 2 and 10/26 again at month 8. The second vaccine dose was obtained by 21/26 (80%) of the patients seen at month 2, and 9/10 (90%) seen at month 8 obtained the third vaccine dose by primary care physicians or ambulant hepatologists. Only 2 patients presented with an anti-HBs-titer >10 IU/L at month 8. CONCLUSIONS Initiation of HBV vaccination during hospitalization and detailed recommendations on subsequent vaccinations in the discharge letter improve previously inadequate vaccination rates in the outpatient setting. Similar measures should be implemented at earlier time points of chronic liver diseases to achieve higher immune response rates.


Dermoscopic Differentiation of Facial Lentigo Maligna from Pigmented Actinic Keratosis and Solar Lentigines.
The differential diagnosis of lentigo maligna (LM) from pigmented actinic keratosis (PAK) and solar lentigines (SL) remains a challenge for clinicians, especially in the early stages of LM when there are no distinctive dermoscopic features. Objective of this study was to evaluate the frequencies of selective dermoscopic criteria in LM, PAK, and SL and to find the specific combination of distinguishing dermoscopic criteria for LM. Dermoscopists blinded to histopathological diagnosis evaluated 42 LM, 107 PAK, and 16 SL for the presence of predefined dermoscopic criteria. The differences in the presence of dermoscopic criteria between LM and others were evaluated with the chi-squared test or Fisher's exact test as appropriate. Multivariate logistic regression analysis with the forward conditional stepwise method were performed and odds ratios and corresponding 95% confidence intervals for LM, PAK, and SL were calculated. LM, PAK, and SL showed many common dermoscopic findings. In multivariate logistic regression analysis, darkening at dermoscopic examination (sevenfold), gray circles (sevenfold), target-like pattern (sixfold), gray rhomboids (sixfold), and slate-gray dots/globules (threefold) represented the strongest predictors of LM, while hyperkeratosis (thirteenfold), white circles (twelvefold), and red rhomboids (sixfold) represented the strongest predictors of PAK. The dermoscopic diagnosis of a given lesion should be based on the presence of the combination of specific dermoscopic criteria rather than a single benign or malignant criterion. Our results suggest that the presence of darkening at dermoscopic examination, gray circles, target-like pattern, gray rhomboids, and slate-gray dots/globules should be considered supportive findings for the diagnosis of early LM.


[Enterobacter cloacae--antimicrobial susceptibility of strains isolated from hospitalized patients in 2004-2005].
The profiles of resistance of ESBL(-) and ESBL(+) strains of Enterobacter cloacae were analysed and compared. 466 Enterobacter cloacae strains isolated from different specimens obtained from patients of big Warsaw hospital in 2004-2005 were investigated. By using the several phenotypic methods 33.5% of strains was identified as ESBL(+). ESBL(+) strains were significantly less susceptible then ESBL(-). These two groups differed mostly in susceptibility to aminoglycosides and fluoroquinolones, e.g. 65.5% of ESBL(-) strains were susceptible to gentamicin, compared to only 25.0% ESBL(+), in case of ciprofloxacin 59.0% of ESBL(-) were susceptible whereas only 25.0% of ESBL(+) strains. The percentage of ESBL(+) grew from 26.4% in 2004 to 40.4% in 2005, whereas a tendency of growing susceptibility to aminoglycosides and fluoroquinolones was noted among all E.cloacae strains. Susceptibility to combinations of piperacillin or ticarcillin with beta-lactamase inhibitor was rather low among ESBL(+) strains.


Synergistic lithium chloride and glial cell line-derived neurotrophic factor delivery for peripheral nerve repair in a rodent sciatic nerve injury model.
Restoring peripheral nerve function after long gap peripheral nerve damage is challenging. Lithium chloride has demonstrated neuroprotective qualities and therefore shows great potential therapeutic benefit for some neurodegenerative diseases. This study examined the synergistic combination of glial cell line-derived neurotrophic factor and lithium chloride and its effect on peripheral nerve regeneration in a rat sciatic nerve injury model. Polycaprolactone conduits with glial cell line-derived neurotrophic factor-loaded double-walled microspheres and local injections of lithium chloride, 1.5 or 2.5 mEq/kg body weight, were examined in a 15-mm rat sciatic nerve defect model. Eighteen Lewis male rats were divided randomly into control, 1.5-, and 2.5-mEq/kg lithium chloride injection groups. As an indicator of recovery, nerve sections were stained with S100, protein gene product 9.5 antibody, and toluidine blue. Nerves stained with S100 and protein gene product 9.5 antibody demonstrated a significantly increased density of Schwann cells and axons in the 2.5-mEq/kg lithium chloride injection-treated groups compared with both the control and 1.5-mEq/kg lithium chloride injection-treated groups (p<0.05). At 6 weeks, histomorphometry revealed a significantly higher fiber density in the middle of the conduit for the 2.5-mEq/kg groups compared with the 1.5-mEq/kg group or the control group. Polycaprolactone nerve guides with glial cell line-derived neurotrophic factor-loaded double-walled microspheres and local injections of lithium chloride, 2.5-mEq/kg, represent a potentially viable guiding material for Schwann cell and axon migration and proliferation for the treatment of peripheral nerve regeneration.


Antitumoural activity of a cytotoxic peptide of Lactobacillus casei peptidoglycan and its interaction with mitochondrial-bound hexokinase.
In a previous study, we reported the cytotoxic activity against various tumour cells of the peptidoglycan of Lactobacillus casei. To isolate the most active components, we performed column-chromatography separation of the peptidoglycan complex and tested the related fractions for their cytotoxic activity. The most active fractions were then lyophilized and the residue was analysed by gas chromatography for its amino acid content and composition. On the basis of the known chemical formula of the basic peptidic component of the peptidoglycan complex of L. casei, a peptide was then synthesized [Europ. (CH-DE-FR-GB) Patent number 1217005; IT number 01320177] and its cytotoxicity was tested against tumoural and normal cells. The synthetic peptide was found to impair the entire metabolism of cultured tumour cells and to restore the apoptotic process. By contrast, normal cells appeared to be stimulated rather than inhibited by the peptide, whereas primary mouse embryo fibroblasts behaved similarly to tumour cells. On the basis of these results, L. casei peptidoglycan fragments and their constituent basic peptide might be applicable as potent antitumour agents.


Protein prenylcysteine analog inhibits agonist-receptor-mediated signal transduction in human platelets.
Signal transduction components, including the Ras superfamily of low molecular weight GTP-binding proteins and the gamma subunits of heterotrimeric G proteins, are reversibly carboxyl methylated at C-terminal prenylcysteine residues. We have previously shown that the prenylcysteine analog N-acetyl-S-trans,trans-farnesyl-L-cysteine (AFC) inhibits carboxyl methylation of these proteins in human platelets. Here we show that concentrations of AFC that inhibit Ras carboxyl methylation (10-50 microM) also block responses to agonists such as ADP, collagen, arachidonic acid, U46619 (a stable analog of prostaglandin H2), thrombin, and guanosine 5'-[gamma-thio]triphosphate. AFC does not inhibit aggregation induced by effectors such as ionomycin, phorbol 12,13-dibutyrate, and bacterial phospholipase C that bypass G proteins to activate platelets at the level of cytosolic Ca2+ concentration and protein kinase C. These findings indicate that AFC inhibits agonist-receptor-mediated signal transduction in human platelets.


Antegrade and retrograde endoscopy for treatment of esophageal stricture.
Total or near-total esophageal stricture results from multiple processes. Traditional treatment with wire cannulation followed by serial dilation is often contraindicated due to poor visualization and the risk of perforation. We seek to demonstrate that combined antegrade and retrograde endoscopy are useful for treatment of total or near-total esophageal strictures. The gastrostomy tube is removed and the tract dilated. A standard endoscope is passed retrograde to the stricture. An antegrade endoscope is advanced until transillumination across the stricture is visualized. A biopsy forceps or needle is used to traverse the stricture in an antegrade fashion. The tract is cannulated with a stiff wire that is then brought out through the gastrostomy site. The stricture is serially dilated. The gastrostomy tube is replaced, and a nasogastric tube is left across the stricture for 3 to 4 weeks. The endoscope is withdrawn and an 18 or 20 Fr gastrostomy tube is left in place. A total of three patients with total esophageal strictures were treated using combined antegrade and retrograde esophagoscopy. All three patients regained the ability to swallow secretions. Importantly, there were no instances of esophageal perforation. This technique has broader application, including combination with minilaparotomy for patients without retrograde access. Further research is needed to determine durability of stricture dilation.


Inequalities in mammography and Papanicolaou test coverage: a time-series study.
Cancer is a serious public issue problem worldwide. In Brazil, breast cancer is the most common type and cervical cancer is the third most frequent among women. To analyze the temporal trend of coverage of mammography and cervical oncotic cytological testing, between 2007 and 2018. Time-series study conducted in the 26 Brazilian state capitals and in the Federal District. A linear regression model was used to estimate trends in coverage of mammography and cervical oncotic cytological testing over the period. The data collection system for Surveillance of Risk and Protection Factors for Chronic Diseases by Telephone Survey (Vigitel) was used. A significant increase in mammography coverage was observed, from 71.1% in 2007 to 78.0% in 2018. There was a trend towards an increase among women with 0 to 8 years of schooling, in all regions of Brazil. Regarding cervical oncotic cytological testing coverage, there was a trend towards stability during the period analyzed, reaching 81.7% in 2018. On the other hand, there was a significant increase in the northern region. There was an improvement in the coverage of these screening examinations, especially regarding mammography. However, it is still necessary to expand their provision, quality and surveillance, aimed towards women's health.


Pseudo-rotation mechanism for fast olefin exchange and substitution processes at orthometalated C,N-complexes of platinum(II).
Bridge splitting in chloroform of the orthometalated chloro-bridged complex [Pt(micro-Cl)(2-Me(2)NCH(2)C(6)H(4))](2)(1), with ethene, cyclooctene, allyl alcohol and phosphine according to 1+ 2L --> 2[PtCl(2-Me(2)NCH(2)C(6)H(4))(L)], where L = C(2)H(4)(3a), C(8)H(14), (3b), CH(2)CHCH(2)OH (3c), and PPh(3)(4a and 4b) gives monomeric species with L coordinated trans or cis to aryl. With olefins the thermodynamically stable isomer with L coordinated cis to aryl is formed directly without an observable intermediate. With phosphine and pyridine, the kinetically controlled trans-product isomerizes slowly to the more stable cis-isomer. Bridge splitting by olefins is slow and first-order in 1 and L, with largely negative DeltaS(++). Substitution of ethene cis to aryl by cyclooctene and allyl alcohol to form 3b and 3c, and substitution of cot from 3b by allyl alcohol to form 3c are first order in olefin and complex, ca. six orders of magnitude faster than bridge cleavage due to a large decrease in DeltaH(++), and with largely negative DeltaS(++). Cyclooctene exchange at 3b is first-order with respect to free cyclooctene and platinum complex. All experimental data for olefin substitution and exchange are compatible with a concerted substitution/isomerization process via a turnstile twist pseudo-rotation in a short-lived labile five-coordinated intermediate, involving initial attack on the labile coordination position trans to the sigma-bonded aryl. Bridge-cleavage reactions of the analogous bridged complexes occur similarly, but are much slower because of their ground-state stabilization and steric hindrance.


Subacute toxicity of anilofos, a new organophosphorus herbicide in male rats: effect on lipid peroxidation and ATPase activity.
Effects of anilofos on lipid peroxidation--an index of oxidative stress, ATPase activity--an integral part of active transport mechanisms for cations, GSH level and GST activity were evaluated in blood (erythrocyte/plasma), brain and liver of male rats after daily oral exposure to 50, 100 or 200 mg/kg for 28 days. None of the doses increased lipid peroxidation. The lowest dose, rather, produced marginally significant decrease in peroxidation in liver. Different doses of anilofos decreased GSH content and activities of GST and ATPases. Inhibition of total ATPase (34-44%) and Na+-K+-ATPase (45-52%) activities was maximum in liver, while that of Mg2+-ATPase (46-56%) was more in erythrocyte. Results indicate that anilofos may not cause oxidative damage to cell membrane in repeatedly exposed animals and may cause neuronal/cellular dysfunction by affecting ionic transport across cell membrane.


Flavonoids and stilbenoids with COX-1 and COX-2 inhibitory activity from Dracaena loureiri.
Fom the stem wood of Dracaena loureiri, a new homoisoflavanone named loureiriol (1) and eight known flavonoid and stilbenoid derivatives, including 5,7-dihydroxy-3-(4-hydroxybenzyl)-4-chromanone (2), 4,4'-dihydroxy-2,6-dimethoxydihydrochalcone (3), 2,4'-dihydroxy-4,6-dimethoxydihydrochalcone (4), 4'-hydroxy-2,4,6-trimethoxydihydrochalcone (5), 4,6,4'-trihydroxy-2-methoxydihydrochalcone (6), 4,3',5'-trihydroxystilbene (7), 4,3'-dihydroxy-5'-methoxystilbene (8) and 4-hydroxy-3',5'-dimethoxystilbene (9) were isolated. These compounds were evaluated for their inhibitory activity against the enzymes cyclooxygenase-1 and cyclooxygenase-2. Potent but non-selective activity was found for the stilbenoids 7-9 (IC(50) 1.29 - 4.92 microM) whereas weak or no activity was observed for the flavonoids 1-6.


Women and migration: the social consequences of gender.
"This paper reviews the literature on the neglected role of women in migration. It argues that focusing on gender and the family can provide the necessary linkage of micro and macro levels of analyses. Striving to contribute to a gendered understanding of the social process of migration, the review organizes the literature along these major issues: How is gender related to the decision to migrate--i.e. what are the causes and consequences of female or male-dominated flows of migration? What are the patterns of labor market incorporation of women immigrants--i.e. what accounts for their participation in the labor force and their occupational concentration? What is the relationship of the public and the private--i.e. what is the impact of work roles on family roles and of the experience of migration on the immigrants themselves? Throughout, the necessity to understand how ethnicity, class, and gender interact in the process of migration and settlement is stressed."


Ring A/B-Modified 17β-Hydroxywithanolide Analogues as Antiproliferative Agents for Prostate Cancer and Potentiators of Immunotherapy for Renal Carcinoma and Melanoma.
Physachenolide C (1) is a 17β-hydroxywithanolide natural product with a unique anticancer potential, as it exhibits potent and selective in vitro antiproliferative activity against prostate cancer (PC) cells and promotes TRAIL-induced apoptosis of renal carcinoma (RC) and poly I:C-induced apoptosis of melanoma cells. To explore the effect of ring A/B modifications of physachenolide C (1) on these biological activities, 23 of its natural and semisynthetic analogues were evaluated. Analogues 4-23 were prepared by chemical transformations of a readily accessible compound, physachenolide D (2). Compound 1 and its analogues 2-23 were evaluated for their antiproliferative activity against PC (LNCaP and 22Rv1), RC (ACHN), and melanoma (M14 and SK-MEL-28) cell lines and normal human foreskin fibroblast (HFF) cells. Most of the active analogues had selective and potent activity in reducing cell number for PC cell lines, some showing selectivity for androgen-independent and enzalutamide-resistant 22Rv1 cells compared to androgen-dependent LNCaP cells. Analogues with IC50s below 5.0 μM against ACHN cells, when tested in the presence of TRAIL, showed a significantly increased ability to reduce cell number, and those analogues active against the M14 and SK-MEL-28 cell lines exhibited enhanced activity when combined with poly I:C. These data provide additional structure-activity relationship information for 17β-hydroxywithanolides and suggest that selective activities of some analogues may be exploited to develop natural products-based tumor-specific agents for cancer chemotherapy.


Role of the Solvation Water in Remote Interactions of Hyperactive Antifreeze Proteins with the Surface of Ice.
Most protein molecules do not adsorb onto ice, one of the exceptions being so-called antifreeze proteins. In this paper, we describe that there is a force pushing an antifreeze protein molecule away from the ice surface when it is not oriented with its ice-binding plane toward the ice and that this pushing force may be also present even when the protein is oriented with its ice-binding plane toward the ice. This force is absent only when certain specific distance criteria are met, regarding the surface of ice and the protein. It acts at early stages of adsorption, prior to the solidification of water between the ice and the protein molecule nearby. We propose the water-originating mechanism of the generation of this force and also the mechanism of remote attachment of an antifreeze molecule to the ice surface. In liquid water, there exist locally favored structures, ordered and of high specific volume. The presence of a protein molecule usually shifts the equilibrium that exists in liquid water toward increasing the number of high-density, disordered structures and diminishing the number of low-density structures. Creation of the locally favored structures may be hampered not only near the non-ice-binding surfaces but also between the ice surface and the protein surface, if the distance between these surfaces does not allow these structures to develop because the available space is not sufficient for their proper formation. This conclusion is supported by the analysis of the mean geometry of a single hydrogen bond, as well as of the hydrogen bond network in the solvation layer and a structural order parameter that characterizes the separation between the first and second solvation shells of a water molecule.


Porphyrin self-assembly at electrochemical interfaces: role of potential modulated surface mobility.
The self-assembly of 5,10,15,20-tetra(4-pyridyl)-21H,23H-porphine (TPyP) on Au(111) electrodes was investigated. The adlayer structure was found to depend on the electrode potential. At positive potentials (>0.5V(SCE)), a disordered layer of TPyP is formed on the Au(111) electrode. STM images showed that the disordered molecules are immobile. At negative potentials (-0.2V(SCE)), however, the molecules are highly mobile and can no longer be imaged by STM, though they remain on the surface. At intermediate potentials (-0.2 to +0.2V(SCE)), the TPyP formed a highly ordered adlayer. Once the ordered adlayer is formed, it persists even after the potential is stepped to higher values (0.5-0.8 V(SCE)). These results can be explained by the role of potential modulated adsorbate-substrate interaction and surface mobility. This suggests the intriguing prospect of using electrode potential to tune surface interactions and to drive surface processes, e.g., molecular self-assembly, in electrochemical systems.


Construction and immunogenicity study of a 297-bp humanized HIV V3 DNA of an approximated last common ancestor in mice.
DNA immunization represents one of the promising HIV-1 vaccine approaches. To overcome the obstacle of genetic variation, we used the last common ancestor (LCA) or "center-of-the-tree" approach to study a DNA fragment of the HIV-1 envelope surrounding the V3 region. A humanized codon of the 297-bp consensus ancestral sequence of the HIV-1 envelope (codons 291-391) was derived from the 80 most recent HIV-1 isolates from the 8 circulating HIV-1 subtypes worldwide. This 297-bp humanized "multi-clade" V3 DNA was amplified by a PCR-based technique. The PCR product was well expressed in vitro whereas the corresponding non-humanized V3 DNA (subtype A/E) could not be expressed. However, both V3 DNA constructs as well as the full-length HIV-1 envelope construct (A/E) were found to be immunogenic in mice by the footpad-swelling assay. Moreover, intracellular and extracellular interferon-gamma could be detected upon in vitro stimulation of spleen cells although the response was relatively weak. Further improvement of our humanized V3 DNA is needed.


Exercise training-induced changes in inflammatory mediators and heat shock proteins in young tennis players.
Heat shock proteins (Hsp) represent proteins' groups, whose protective function, may be induced by heat, reactive oxygen species, cytokines etc. We evaluated blood levels of Hsp27 and Hsp70, and their relation to skeletal muscle damage and inflammation in young tennis players before and after the conditioning camp. Blood samples were collected directly after tournament season, 3-day rest and 14-day conditioning camp that followed. Hydrogen peroxide (H2O2) demonstrated the highest concentration directly after tournament season, which significantly decreased at camp's end. The pro-inflammatory cytokines IL-1β and TNFα decreased, whereas anti-inflammatory cytokines IL-6 and IL-10 increased after 3d rest and 14d camp. Hsp27 increased after 3d rest and remained so after 14d camp, while Hsp70 decreased from baseline to camp's completion. Hsp27 and Hsp70 correlated significantly with H2O2, IL-1β and TNFα. Muscle damage, observed as creatine kinase (CK) activity changes, increased after 14d camp similarly to Hsp27 and anti-inflammatory cytokines IL-6 and IL-10. Obtained data allows to conclude that decrease of Hsp27 and increase in pro-inflammatory cytokines could be a good indicator of overreaching. Reverse tendencies in these proteins may verify accuracy of conditioning camp. Finally, this training program caused an increase in the anti-inflammatory cytokines concentrations, improving individual status of recovery. Key PointsThe study demonstrating low grade inflammation-induced by the tournament season in young tennis player.Three days of active rest stimulated the anti-inflammatory response via rise of Hsp27 and anti-inflammatory cytokine IL-10.Observed decrease of blood Hsp70 may support mental recovery.Thirteen-day appropriate training program led to maintaining an immunological response balance.


Perceived benefits and barriers to physical activity among older Latina women.
Evidence of the benefits of physical activity in the health of aging women continues to grow but questions remain about the factors that influence these women's ability to engage in this behavior. The authors used a qualitative design to describe the social and culture-specific perceived benefits and barriers to physical activity among 143 Latina women, ages 40 to 79. Content analysis of these women's responses revealed that perceived benefits (health promotion, physical fitness, improved roles) and barriers (time constraints of women's roles, personal health, internal and external factors) function as competing elements that may explain physical inactivity. Health care providers should emphasize overcoming barriers and promote perceived benefits as clinical interventions that may pose the greatest potential to increase physical activity among aging Latina women. This emphasis holds promise as a feasible and effective primary care intervention for achieving increased physical-activity-related health benefits.


Coronary computed tomographic angiographic findings in patients with Kawasaki disease.
Kawasaki disease (KD) is the leading cause of acquired coronary disease in children and may lead to subsequent myocardial ischemia and infarction. Because coronary computed tomographic angiography (CTA) is the most sensitive noninvasive test in patients with atherosclerosis, the aim of this study was to retrospectively evaluate coronary CTA performed in patients with KD for aneurysm, stenosis, and calcified and noncalcified coronary artery disease (CAD). Clinical histories and prior stress and imaging test results were reviewed. Thirty-two patients underwent coronary CTA for KD, and 385 coronary segments were evaluated. Twenty-three of 32 patients had ≥1 diseased coronary segment. There were 20 aneurysms, 7 lesions, and 75 segments (20%) with nonobstructive CAD (16% noncalcified, 2% calcified, and 2% mixed). All nonobstructive and obstructive CAD was in patients with histories of acute-phase coronary artery dilatation or aneurysm (echocardiographic z score 4 to 44), and were almost always associated with normal stress imaging test results on follow-up. No lesion or CAD was found in coronary computed tomographic angiographic studies performed in a control group referred for other indications (n = 32, 422 segments evaluated). The median coronary computed tomographic angiographic dose-length product was 59 mGy cm (interquartile range 32 to 131), the median unadjusted radiation dose was 0.8 mSv (interquartile range 0.4 to 1.8), and the median age- and size-adjusted radiation dose was 1.3 mSv (interquartile range 0.7 to 2.3). In conclusion, high-risk patients with histories of KD had nonobstructive and obstructive CAD not visualized by other noninvasive imaging tests. In properly selected high-risk patients with KD, coronary CTA may identify a subset at increased risk for future coronary pathology who may benefit from medical therapy.


Contribution of membrane-binding and enzymatic domains of penicillin binding protein 5 to maintenance of uniform cellular morphology of Escherichia coli.
Four low-molecular-weight penicillin binding proteins (LMW PBPs) of Escherichia coli are closely related and have similar DD-carboxypeptidase activities (PBPs 4, 5, and 6 and DacD). However, only one, PBP 5, has a demonstrated physiological function. In its absence, certain mutants of E. coli have altered diameters and lose their uniform outer contour, resulting in morphologically aberrant cells. To determine what differentiates the activities of these LMW PBPs, we constructed fusion proteins combining portions of PBP 5 with fragments of other DD-carboxypeptidases to see which hybrids restored normal morphology to a strain lacking PBP 5. Functional complementation occurred when truncated PBP 5 was combined with the terminal membrane anchor sequences of PBP 6 or DacD. However, complementation was not restored by the putative carboxy-terminal anchor of PBP 4 or by a transmembrane region of the osmosensor protein ProW, even though these hybrids were membrane bound. Site-directed mutagenesis of the carboxy terminus of PBP 5 indicated that complementation required a generalized amphipathic membrane anchor but that no specific residues in this region seemed to be required. A functional fusion protein was produced by combining the N-terminal enzymatic domain of PBP 5 with the C-terminal beta-sheet domain of PBP 6. In contrast, the opposite hybrid of PBP 6 to PBP 5 was not functional. The results suggest that the mode of PBP 5 membrane anchoring is important, that the mechanism entails more than a simple mechanical tethering of the enzyme to the outer face of the inner membrane, and that the physiological differences among the LMW PBPs arise from structural differences in the DD-carboxypeptidase enzymatic core.


Observation of broadband self-amplified spontaneous coherent terahertz synchrotron radiation in a storage ring.
Bursts of coherent synchrotron radiation at far-infrared and millimeter wavelengths have been observed at several storage rings. A microbunching instability has been proposed as the source for the bursts. However, the microbunching mechanism has yet to be elucidated. We provide the first evidence that the bursts are due to a microbunching instability driven by the emission of synchrotron radiation in the bunch. Observations made at the Advanced Light Source are consistent with the values predicted by the proposed microbunching model. These results demonstrate a new instability regime for high energy synchrotron radiation sources and could impact the design of future sources.


A comparison of survival at different degrees of hemorrhagic shock in germ-free and germ-bearing rats.
We have previously reported superior survival after one level of hemorrhagic shock in germ-free (GF) rats compared with germ-bearing (GB) rats. The objective of this study was to determine the effect of the GF state on survival at different degrees of hemorrhagic shock. GF and GB rats were bled to a mean arterial blood pressure of 30 mmHg. Shock was terminated after 10, 20, 40, or 80% of the maximum shed blood volume was reabsorbed spontaneously. Both shock time and time to decompensation were significantly longer in GF rats (p < .05). Comparative survival was greater for GF rats at most levels of shock (p < .01). This superiority in survival was greatest at moderate shock levels and decreased at severe shock levels. There may be several reasons for the increased tolerance of GF animals to hemorrhagic shock such as metabolic or immunologic variations. It is hard to avoid the fact, however, that the most notable difference between the GF and GB rat is the presence or absence of bacteria.


Spatial distribution of buffy-tufted-ear (Callithrix aurita) and invasive marmosets (Callithrix spp.) in a tropical rainforest reserve in southeastern Brazil.
We investigated the spatial distribution of native and invasive marmoset species (Callithrix), as well as their hybrids, in the Serra dos Órgãos National Park (PARNASO) and surrounding area in Rio de Janeiro state, Brazil. To estimate occupancy and the detection probability, we surveyed 56 sites within the park and 52 sites outside its limits using vocal playbacks, as well as by interviewing local residents in the surrounding area. We estimated the occupancy and detection probability of Callithrix aurita and the observed groups composed of Callithrix jacchus, Callithrix penicillata, and their hybrids. We also recorded the presence or absence of mixed groups of native and exotic species, and their hybrids. We recorded similar occupancy rates and detection probabilities for both native and invasive species within the national park. C. aurita was found more often within the areas of the park located furthest from access roads and with the least human interference, while invasive species were more likely to be found along the edge of the park and in areas with greater human interference. In the area surrounding the park, invasive marmosets were recorded at seven sites, and a mixed group of native and invasive marmosets was observed at one site, but non-hybrid C. aurita groups were not recorded. The occupancy probability of C. aurita in the study area is relatively low, which may indicate a low population density, with groups restricted to a small region within the PARNASO in the proximity of groups of invasive marmosets.


Fungal Infection among Patients with Some Hematopoietic Disorders.
With the increasing number of immune compromised patients suffering from hematological disorders, invasive fungal infections have emerged as a major cause of morbidity and mortality among these patients. This study evaluated the use of conventional blood culture and PCR techniques in the isolation and identification of fungi from patients with different hematopoietic disorders. Ninety blood samples were tested by both techniques, conventional blood culture detected fungal infections in only five (5.6%) samples, which were further identified to be of Candida spp. While PCR technique detected fungal infections in 56 (62.2%) samples, which were further subjected to nested PCR technique giving final identification of Candida spp. infections in 36 (40%) samples and Aspergillus spp. in 16 (17.8) samples. The sensitivity of PCR technique was 100% and the specificity ranged from 40% to 100%. Thus, the use of nested PCR technique was an efficient and sensitive method for detection of invasive fungal infections among patients with hematopoietic disorders.


Ephrin b2 receptor and microsatellite status in lymph node-positive colon cancer survival.
Ephrin B2 receptor (EphB2) is a target of the canonical wnt pathway implicated in colorectal carcinogenesis, and its down-regulation may be associated with adverse prognosis. We evaluated its prognostic value in resected colon cancer stratified by microsatellite status and other clinicopathologic characteristics. We identified all cases of resected stage III colon cancer from 1995 to 2009 managed in the Capital Health district of Nova Scotia. Tissue microarrays were constructed and immunohistochemistry (IHC) for tumor EphB2 staining assigned into quartiles. Microsatellite status was evaluated by IHC for MutL homolog 1 (MLH1) and MutS homolog 2 (MSH2). Microsatellite stable tumors were defined as both MLH1/MSH2 (+/+); tumors staining otherwise were classified with microsatellite instability (MSI-H). Primary and secondary outcomes were disease-free survival (DFS) and overall survival (OS), respectively. We identified 159 cases with sufficient tissue for microarray analysis having a median follow-up of 3.47 years (range, 0.14-14). Median age was 61, 52% were male, 40% had an event, and 29% died. MSI-H was present in 18 (13%). Univariate analysis of EphB2 expression on DFS and OS showed a hazard ratio (HR) of 2.00 (P = .01) and 2.14 (P = .03), respectively. Multivariate analysis of EphB2 expression on DFS and OS showed an HR of 2.24 and 2.23, respectively, with tumor IHC ≤ 50%. In this cohort, decreased EphB2 expression was an independent prognostic factor for recurrence and death and may have prognostic relevance in tumors with MSI-H. However, this would require prospective validation in a larger study.


Life-threatening haematemesis associated with clozapine: a case report and literature review.
Clozapine is regarded as a second-line and in some cases last-line antipsychotic known for its common life-threatening side effects, such as agranulocytosis, constipation and cardiomyopathies, but rarely haematemesis. We report a case of severe haemetemesis in a chronic schizophrenic patient managed with clozapine. The patient was a 46-year-old male being managed for chronic schizophrenia with treatment resistance who developed sudden severe haematemesis following commencement of clozapine for 6 weeks. The patient had 1.1 l of blood transfusion. The relevant literature is reviewed. Clozapine can be associated with a life-threatening haematemesis. Psychiatrists and other medical specialists need to be alert to the fatality of clozapine-induced haematemesis in the treatment of psychiatric disorders.


2D full-field displacement and vibration measurements of specularly reflecting surfaces by two-beam common-path digital holography.
A new, to the best of our knowledge, configuration of common-path off-axis digital holography is proposed for simultaneous evaluation of out-of-plane and in-plane displacements of the vibrating object. The object is illuminated from two different directions, and each illumination interferes with its corresponding reference beam generated near the object, resulting in two independent holograms that are spatially multiplexed in a single camera image. Two multiplexed holograms, at undeformed and deformed states of the object, are recorded and processed to obtain the out-of-plane and in-plane displacements simultaneously. The proposed digital holographic system has the advantage of a simple and compact optical setup, is less sensitive to environmental disturbances, and has high temporal phase stability. The two-dimensional (z,x) full-field amplitude and phase vibration analysis of a perfect specularly reflecting surface are also demonstrated by the proposed holographic system. The experimental results authenticate the feasibility of the proposed system and reveal its unique advantages. The proposed digital holographic system, owing to simple and compact geometry and providing several advantages over other two-channel holographic systems, may find a wide range of applications in investigating real-time dynamic phenomena.


Multiple novel promoter-architectures revealed by decoding the hidden heterogeneity within the genome.
An important question in biology is how different promoter-architectures contribute to the diversity in regulation of transcription initiation. A step forward has been the production of genome-wide maps of transcription start sites (TSSs) using high-throughput sequencing. However, the subsequent step of characterizing promoters and their functions is still largely done on the basis of previously established promoter-elements like the TATA-box in eukaryotes or the -10 box in bacteria. Unfortunately, a majority of promoters and their activities cannot be explained by these few elements. Traditional motif discovery methods that identify novel elements also fail here, because TSS neighborhoods are often highly heterogeneous containing no overrepresented motif. We present a new, organism-independent method that explicitly models this heterogeneity while unraveling different promoter-architectures. For example, in five bacteria, we detect the presence of a pyrimidine preceding the TSS under very specific circumstances. In tuberculosis, we show for the first time that the spacing between the bacterial 10-motif and TSS is utilized by the pathogen for dynamic gene-regulation. In eukaryotes, we identify several new elements that are important for development. Identified promoter-architectures show differential patterns of evolution, chromatin structure and TSS spread, suggesting distinct regulatory functions. This work highlights the importance of characterizing heterogeneity within high-throughput genomic data rather than analyzing average patterns of nucleotide composition.


Kojic acid protects C57BL/6 mice from gamma-irradiation induced damage.
The radioprotective effects of a single administration of kojic acid (KA) against ionizing radiation were evaluated via assessment of 30-day survival and alterations of peripheral blood parameters of adult C57BL/6 male mice. The 30-day survival rate of mice pretreated with KA (75 or 300 mg/kg body weight, KA75 or KA300) subcutaneously 27 h prior to a lethal dose (8 Gy, 153.52 cGy/min) of gamma irradiation was higher than that of mice irradiated alone (40% or 60% vs 0%). It was observed that the white blood cell (WBC) count/the red blood cell (RBC) count, haemoglobin content, haematocrit and platelet count of mice with or without KA pretreatment as exposed to a sub-lethal dose (4 Gy, 148.14 cGy/min) of gamma irradiation decreased maximally at day 4/day 8 post-irradiation. Although the initial WBC values were low in KA300 or WR-2721 (amifostine) groups, they significantly recovered to normal at day 19, whereas in the control group they did not. The results from the cytotoxicity and cell viability assays demonstrated that KA could highly protect Chinese hamster ovary (CHO) cells against ionizing radiation with low toxicity. In summary, KA provides marked radioprotective effects both in vivo and in vitro.


Plasma thiol status is altered in children with mitochondrial diseases.
This study was undertaken to investigate thiol metabolism as a marker of oxidative stress and antioxidative defence capacity in a cohort of children with biochemically and/or genetically confirmed mitochondrial disease. Previous studies suggest that lower glutathione levels, which have been shown to further compromise mitochondrial function, may occur in these diseases. Better understanding of the pathogenesis of mitochondrial diseases is important in order to improve their treatment. We studied plasma and erythrocyte glutathione and cysteine levels, the activities of erythrocyte glutathione peroxidase (GPx), glutathione reductase (GR), glucose 6-phosphate dehydrogenase G6PDH) and glutathione S-transferase (GST), as well as the levels of erythrocyte thiobarbituric acid-reactive species (TBA-RS) and protein carbonyls in 10 children with a biochemical and/or genetic diagnosis of mitochondrial disease and six controls. Levels of reduced cysteine (CYSH) as well as reduced to oxidised cysteine ratio were lower in plasma of patients with mitochondrial diseases (p = 0.008 and p = 0.02, respectively). Plasma levels of reduced glutathione (GSH) were low in patients with mitochondrial diseases, mostly below the detection limit. We did not detect significant differences in erythrocyte thiols or glutathione-related enzyme activities. Plasma thiols and their redox state are altered in patients with mitochondrial diseases, suggesting an increase in oxidative stress and depletion of antioxidant supplies. If confirmed in further studies, this relative thiol deficiency could be an important factor in the pathophysiology of mitochondrial diseases.


Aquabirnaviruses isolated from marine organisms form a distinct genogroup from other aquabirnaviruses.
A phylogenetic tree of aquabirnaviruses, including marine birnaviruses (MABV) and infectious pancreatic necrosis virus (IPNV), was developed based on the nucleotide sequences and deduced amino acid sequences of the polyprotein and VP5 genes of genomic segment A. In the polyprotein of MABV strains, the amino acid sequences were very similar, with identities of 98.3-99.7%. Twenty-one unique amino acid residues were found in the deduced amino acid sequences of the polyprotein gene of MABV strains. The phylogenetic tree based on the nucleotide sequence of genomic segment A and polyprotein sequences showed that 31 aquabirnavirus strains were clustered into seven genogroups. All MABV strains isolated in Japan and Korea were clustered into one genogroup which was distinct from other aquabirnaviruses. The seventh genogroup containing all MABV strains showed amino acid sequence similarities of 80.7-90.6% with other genogroups. In VP5, four unique residues were found in MABV strains when compared with IPNV strains. The MABV strains exhibited amino acid sequence similarities of 63.9-86.4% with IPNV strains. The amino acid sequences of VP5 were conserved among MABV strains, but differed from those of IPNV strains. The MABV strains isolated from different host species and different geographical areas were very similar to each other, suggesting that the MABV are distinct from the other genogroups.


Causes of Hospital Violence, Characteristics of Perpetrators, and Prevention and Control Measures: A Case Analysis of 341 Serious Hospital Violence Incidents in China.
Objective: Hospital violence remains a global public health problem. This study aims to analyze serious hospital violence causes in China and the characteristics of perpetrators. It likewise seeks to understand frontline personnel's needs and put forward targeted suggestions. Methods: Serious hospital violence cases from 2011 to 2020 in the China Judgment Online System (CJOS) were selected for descriptive statistical analysis. A total of 72 doctors, nurses, hospital managers, and security personnel from 20 secondary and tertiary hospitals in China were selected for semi-structured interviews. Results: Of the incidents, 62.17% were caused by patients' deaths and dissatisfaction with their treatment results. Moreover, it was found that out-of-hospital disputes (11.14%) were also one of the main reasons for serious hospital violence. The perpetrators were mainly males (80.3%), and had attained junior high school education or lower (86.5%). Furthermore, most of them were family members of the patients (76.1%). Healthcare workers urgently hope that relevant parties will take new measures in terms of legislation, security, and dispute handling capacity. Conclusion: In the past 10 years, serious hospital violence's frequency in China has remained high. Furthermore, their harmful consequences are more serious. The causes of hospital violence are diverse, and the characteristics of perpetrators are obvious. Frontline healthcare workers urgently need relevant parties to take effective measures in terms of legislation, security, and dispute handling capacity, to prevent the occurrence of violence and protect medical personnel's safety.


Expression of Ascorbic Acid Oxidase in Zucchini Squash (Cucurbita pepo L.).
The expression of ascorbic acid oxidase was studied in zucchini squash (Cucurbita pepo L.), one of the most abundant natural sources of the enzyme. In the developing fruit, specific activity of ascorbic acid oxidase was highest between 4 and 6 days after anthesis. Protein and mRNA levels followed the same trend as enzyme activity. Highest growth rate of the fruit occurred before 6 days after anthesis. Within a given fruit, ascorbic acid oxidase activity and mRNA level were highest in the epidermis, and lowest in the central placental region. In leaf tissue, ascorbic acid oxidase activity was higher in young leaves, and very low in old leaves. Within a given leaf, enzyme activity was highest in the fast-growing region (approximately the lower third of the blade), and lowest in the slow-growing region (near leaf apex). High expression of ascorbic acid oxidase at a stage when rapid growth is occurring (in both fruits and leaves), and localization of the enzyme in the fruit epidermis, where cells are under greatest tension during rapid growth in girth, suggest that ascorbic acid oxidase might be involved in reorganization of the cell wall to allow for expansion. Based on the known chemistry of dehydroascorbic acid, the end product of the ascorbic acid oxidase-catalyzed reaction, we have proposed several hypotheses to explain how dehydroascorbic acid might cause cell wall "loosening."


Ligand specificity of brain lipid-binding protein.
Brain lipid-binding protein (BLBP) is a member of the fatty acid-binding protein (FABP) family. Although BLBP expression in the developing central nervous system is complex, a close correlation between its expression and radial glial differentiation has been observed. Furthermore, antibodies to BLBP can block glial cell differentiation in mixed primary cell cultures. Here we describe the ligand binding properties of BLBP. The binding affinities of BLBP for oleic acid (Kd approximately 0.44 microM) and arachidonic acid (Kd approximately 0.25 microM) are similar to those reported for other FABPs, but BLBP does not bind to palmitic acid or arachidinic acid. These and other experiments establish that BLBP has a strong preference for binding long chain polyunsaturated fatty acids. A probable in vivo ligand for BLBP is docosahexaenoic acid (DHA), since its binding affinity (Kd approximately 10 nM) is the highest yet reported for an FABP/ligand interaction, exceeding even the affinity of retinoic acid for its binding proteins. Furthermore, the requirement of DHA for nervous system development and the coincident expression of BLBP during these developmental stages suggest that the physiologic role of BLBP may involve DHA utilization. Finally, we present a structural model of BLBP/DHA interaction that provides insight into both the structural characteristics important for ligand binding and the effects of specific mutations upon BLBP/ligand interactions.


Multiple attentional control settings influence late attentional selection but do not provide an early attentional filter.
When one is responding to targets containing a specific feature, non-predictive peripheral cues that share this feature lead to faster responses to the target, while cues that do not contain the target feature effectively are ignored, providing evidence for the role of attentional control settings (ACSs) in the contingent capture hypothesis. It is unclear, however, at what stage of processing multiple ACSs are implemented. We took advantage of the excellent temporal resolution of electroencephalography to demonstrate that the maintenance of multiple ACSs influences later stages of attentional selection rather than providing an early attentional filter. N2pc analyses for cues and targets revealed a similar degree of spatial capture for any peripheral cue, regardless of control settings, with target P3s reflecting the application of the ACS color contingencies.


Development and Characterisation of the Imiquimod Poly(2-(2-methoxyethoxy)ethyl Methacrylate) Hydrogel Dressing for Keloid Therapy.
The imiquimod-poly(2-(2-methoxyethoxy)ethyl methacrylate) hydrogel (poly(MEO₂MA) hydrogel) dressing was developed for the keloid therapy application. Four groups of the hydrogels, including the imiquimod-poly(MEO₂MA) hydrogel, crosslinked with 0.2 mol %, 0.4 mol %, 0.6 mol %, and 0.8 mol % of di(ethylene glycol) dimethacrylate cross-linker (DEGDMA), were synthesised and characterised for fabricating the imiquimod-poly(MEO₂MA) hydrogel pad. The lower critical solution temperature (LCST) of the poly(MEO₂MA) hydrogel was measured at approximately 28 °C and was used as a trigger to control the imiquimod loading and release. The loaded amounts of the imiquimod in the poly(MEO₂MA) hydrogel, crosslinked with 0.2 mol % and 0.8 mol % of DEGDMA, were about 27.4 μg and 14.1 μg per 1 mm³ of the hydrogel, respectively. The imiquimod-release profiles of two samples were characterised in a phosphate buffered saline (PBS) solution at 37 °C and the released imiquimod amount were about 45% and 46% of the total loaded imiquimod. The Cell Counting Kit-8 (CCK-8) assay was utilised to analyse the cell viability of keloid fibroblasts cultured on the samples of imiquimod-poly(MEO₂MA) hydrogel, crosslinked with 0.2 mol % and 0.8 mol % of DEGDMA. There was around a 34% decrease of the cell viabilities after 2 days, compared with the pure-poly(MEO₂MA) hydrogel samples. Therefore, the developed imiquimod-poly(MEO₂MA) hydrogel dressing can affect the proliferation of keloid fibroblasts. It should be possible to utilise the hydrogel dressing for the keloid therapy application.


The Destructive Power of Circumstance on Concurrent Poliomyelitis and Osteomyelitis.
The current method in dealing with pediatric hip pathologies is to avoid joint replacement unless absolutely necessary. This decision is typically made on a scientific basis. However, this is not necessarily the correct approach in all cases as demonstrated in the following case report where social issues is an extremely important variable to be considered when deciding on surgical intervention. This case report is the first of its kind to highlight the pathology mentioned and the social determinants that were a causative agent in its development. This case revolves around a 46-year-old Caucasian gentleman who presented in clinic enquiring about the possibility of a hip arthroplasty. He aimed to improve his ability to work after the cessation of state benefits. A retrospective look at his history indicated a number of social factors lead to poor outcomes and the resulting downward spiral causing his current presentation. The effect of social circumstances on the medical and surgical outcomes is highlighted in this report with a look into avascular necrosis and resulting natural arthrodesis. Both of these presentations were secondary to chronic and untreated childhood osteomyelitis of the right hip in conjunction with poliomyelitis development. The dual involvement of poliomyelitis and osteomyelitis to the hip joint no doubt had a massive impact on final pathology and is a rarely documented phenomenon. From an early age, this patient has faced difficulties integrating in society by factors that were out of his control. By the time he was old enough to lead an independent life, the damage had been inflicted both medically and psychologically through his earlier ordeals. By writing this up, it is hoped that clinicians will recognize the debilitating holistic effect that mechanical joint pathologies and neurological deficiencies can have on patients' lives with the view to reopen the discussion on when it is appropriate to surgically intervene in hip pathologies in children and especially where osteomyelitis and poliomyelitis are involved.


Daily Intake of Trehalose Is Effective in the Prevention of Lifestyle-Related Diseases in Individuals with Risk Factors for Metabolic Syndrome.
We previously performed animal studies that suggested that trehalose potentially prevents the development of metabolic syndrome in humans. To evaluate this possibility, we examined whether trehalose suppressed the progression of insulin resistance in a placebo-controlled, double-blind trial in 34 subjects with a body mass index (BMI) ≥23. The subjects were divided into two groups and were assigned to ingest either 10 g/d of trehalose or sucrose with meals for 12 wk. During the study, body composition and blood biochemical parameters were measured at week 0, 8, and 12. These parameters were also measured 4 wk after the end of intake to confirm the washout of test substances. In the trehalose group, blood glucose concentrations after a 2-h oral glucose tolerance test significantly decreased following 12 wk of intake in comparison with baseline values (0 wk). When a stratified analysis was performed in the subjects whose percentage of truncal fat approached the high end of the normal range, the change in body weight, waist circumference, and systolic blood pressure were significantly lower in the trehalose group than in the sucrose group. Our data indicated that a daily intake of 10 g of trehalose improved glucose tolerance and progress to insulin resistance. Furthermore, these results suggested that trehalose can potentially reduce the development of metabolic syndrome and associated lifestyle-related diseases, such as type 2 diabetes.


Real-time intersession and interrater reliability of the functional movement screen.
The purpose of this study was to examine the real-time intersession and interrater reliability of the functional movement screen (FMS). The overall study consisted of 19 volunteer civilians (12 male, 7 female). The intersession reliability consisted of 12 men and 7 women, whereas 10 men and 6 women participated in the interrater reliability test session. Two raters (A and B) were involved in the interrater reliability aspect of this study. The FMS includes 7 tests: deep squat (DS), hurdle step (HS), in-line lunge (IL), shoulder mobility (SM), active straight leg raise (ASLR), trunk stability push-up (TSPU), and rotary stability (RS). Researchers analyzed the data via intraclass correlation (ICC). To determine the reliability of the intersession scoring of the FMS and the intrasession interrater scoring of the FMS a 2-way mixed effects model intraclass correlation coefficient (ICC(3,1)) was used for the continuous data, whereas a weighted Cohen's kappa (κ) was used for the categorical data. The dependent variables were FMS total score (0-21 scale) and associated tests were DS, HS, IL, SM, ASLR, TSPU, and RS. Intersession reliability (ICC, SEM) and κ were as follows: FMS total score (0.92, 0.51), DS (κ = 0.69), HS (κ = 0.16), IL (κ = 0.69), SM (κ = 0.84), ASLR (κ = 0.69), TSPU (κ = 0.77), and RS (no covariance). Interrater reliability (ICC, SEM) and κ were as follows: FMS total score (0.98, 0.25), DS (κ = 1.0), HS (κ = 0.33), IL (κ = 0.88), SM (κ = 0.90), ASLR (κ = 0.88), TSPU (κ = 0.75), and RS (no covariance). The FMS total scores displayed high intersession and interrater reliabilities. Finally, with the exception of HS, all tasks displayed moderate to high intersession reliability and good to high interrater reliability.


Molecular genetic analysis of an SNF2/brahma-related gene in Tetrahymena thermophila suggests roles in growth and nuclear development.
We used a reverse genetic approach to identify three members of the SNF2 superfamily of chromatin remodeling genes in the ciliated protozoan Tetrahymena thermophila in order to investigate possible functions of ATP-dependent chromatin remodeling factors in growth and nuclear development. Comparative sequence analysis of the gene product of the Tetrahymena brahma-related gene (TtBRG1) indicates it is a member of the SNF2/BRM subgroup of the SNF2 superfamily. Northern analysis suggests that TtBRG1 has roles in growth and nuclear development in Tetrahymena. Indirect immunofluorescence analysis during nuclear development indicates that TtBrg1p localizes to both the parental and developing macronucleus of Tetrahymena during the time period corresponding to genome rearrangements. We generated germ line knockout heterokaryons for TtBRG1 and demonstrated that expression of the gene is required to complete nuclear development of Tetrahymena. In addition, the formation of distinct Pdd1p-containing structures is disturbed during the late stages of conjugation in TtBRG1 germ line knockout heterokaryons. We discuss these results in light of possible roles of SNF2-related proteins in growth and nuclear development of Tetrahymena.


Red blood cell alloimmunization in multitransfused patients in a tertiary care center in Western India.
To investigate the seroprevalence and specificity of red blood cell (RBC) antibodies in multitransfused patients, in whom the risk of alloimmunization is especially high. We conducted a retrospective study on blood specimens from 200 multitransfused patients. We evaluated all specimens for alloimmunization using various immunohematological tests via the column agglutination technique. The overall prevalence of RBC alloantibodies was 5.5%. Of the 11 specific types of alloantibodies identified, most (72.7%) belonged to the Rh blood group system, followed by the S, M, and Lewis blood group systems (9.1% each). Most alloantibodies were of the Rh blood group specificity. To improve the quality of blood supplied, especially to patients with thalassemia, we recommend that Rh phenotyped, cross-match-compatible blood should be issued to prevent complications such as acute and delayed hemolytic reactions.


Documented Viral Illness at the Time of Splenic Sequestration Does Not Affect the Odds of Recurrence in Children With Sickle Cell Disease.
This retrospective study investigates the relationship between the presence of a documented viral infection in children with sickle cell disease during their first splenic sequestration crisis and their odds of recurrence. Forty-eight children were admitted to our hospital between 2008 and 2018 with a splenic sequestration crisis. Thirty-six had respiratory viral panels done on admission, 13 of whom were positive. Two additional children were diagnosed with parvovirus B19 infection by serology. The recurrence rate was 52% (17/33) for those deemed negative for viral illness compared with 33% (5/15) among those with a positive documented viral illness, which was not statistically different (P=0.35). HbSC genotype decreased, and reticulocytosis increased the odds of recurrence. Further research is needed to substantiate these findings.


Taphonomic and Diagenetic Pathways to Protein Preservation, Part I: The Case of Tyrannosaurus rex Specimen MOR 1125.
Many recent reports have demonstrated remarkable preservation of proteins in fossil bones dating back to the Permian. However, preservation mechanisms that foster the long-term stability of biomolecules and the taphonomic circumstances facilitating them remain largely unexplored. To address this, we examined the taphonomic and geochemical history of Tyrannosaurus rex specimen Museum of the Rockies (MOR) 1125, whose right femur and tibiae were previously shown to retain still-soft tissues and endogenous proteins. By combining taphonomic insights with trace element compositional data, we reconstruct the postmortem history of this famous specimen. Our data show that following prolonged, subaqueous decay in an estuarine channel, MOR 1125 was buried in a coarse sandstone wherein its bones fossilized while interacting with oxic and potentially brackish early-diagenetic groundwaters. Once its bones became stable fossils, they experienced minimal further chemical alteration. Comparisons with other recent studies reveal that oxidizing early-diagenetic microenvironments and diagenetic circumstances which restrict exposure to percolating pore fluids elevate biomolecular preservation potential by promoting molecular condensation reactions and hindering chemical alteration, respectively. Avoiding protracted interactions with late-diagenetic pore fluids is also likely crucial. Similar studies must be conducted on fossil bones preserved under diverse paleoenvironmental and diagenetic contexts to fully elucidate molecular preservation pathways.


Knockdown of macrophage migration inhibitory factor disrupts adipogenesis in 3T3-L1 cells.
Obesity is a condition in which adipose tissue mass is expanded. Increases in both adipocyte size and number contribute to enlargement of adipose tissue. The increase in cell number is thought to be caused by proliferation and differentiation of preadipocytes. Macrophage migration inhibitory factor (MIF) is expressed in adipocytes, and intracellular MIF content is increased during adipogenesis. Therefore, we hypothesized that MIF is associated with adipocyte biology during adipogenesis and focused on the influence of MIF on adipogenesis. To examine the effects of MIF on adipocytes, MIF expression in 3T3-L1 preadipocytes was inhibited by RNA interference, and cell differentiation was induced by standard procedures. The triglyceride content of MIF small interfering RNA (siRNA)-transfected 3T3-L1 cells was smaller than that of nonspecific siRNA-transfected cells. In addition, MIF knockdown apparently abrogated increases in adiponectin mRNA levels during differentiation. Gene expression of peroxisome proliferator-activated receptor (PPAR)gamma, CCAAT/enhancer binding protein (C/EBP)alpha, and C/EBPdelta decreased with MIF siRNA transfection, but C/EBPbeta expression increased. Cell number and incorporation of 5-bromo-2-deoxyuridine into cells decreased from 1-3 d and from 14-20 h, respectively, after induction of differentiation in MIF siRNA-transfected cells, thus suggesting that MIF siRNA inhibits mitotic clonal expansion. Taken together, these results indicated that MIF regulates differentiation of 3T3-L1 preadipocytes, at least partially, through inhibition of mitotic clonal expansion and/or C/EBPdelta expression.


Optimization of Microwave-Assisted Green Method for Enhanced Solubilization of Water-Soluble Curcuminoids Prepared Using Steviol Glycosides.
In this study, the optimization and modeling of microwave-assisted extraction (MAE) of water-soluble curcuminoids prepared using novel steviol glycosides (SGs) was carried out using four independent process variables at varying levels-X1: microwave power (50-200 W), X2: stevioside concentration (50-200 mg/mL), X3: curcumin concentration (20-200 mg/mL), and X4: time (1-10 min)-in response surface methodology configuration. Moreover, the effects of stevioside, as the most cost-effective natural solubilizer, were also evaluated. The water solubility of curcuminoids increased from 11 to 1320 mg/L with the addition of stevioside as a natural solubilizer. Moreover, microwave heating synergistically with stevioside addition significantly (p < 0.05) increased the solubility up to 5400 mg/L. Based on the results, the optimum conditions providing the maximum solubilization of 16,700 mg/L were 189 W microwave power, 195 g/L stevioside concentration, 183 g/L curcuminoid concentration, and 9 min of incubation time. Moreover, MAE of curcuminoids using SGs might render a significant advantage for its wide-scale application to solubilizing the multitude of insoluble functional flavonoids in fruits, plants, and food materials.


Systemic arterial embolization in patients with hemoptysis: initial experience with ethylene vinyl alcohol copolymer in 15 cases.
The purpose of this study was a preliminary evaluation of the use of ethylene vinyl alcohol copolymer to treat patients with hemoptysis of systemic arterial origin. We reviewed the cases of 15 consecutively registered patients (10 men, five women; mean age, 53.7 years) who underwent endovascular administration of ethylene vinyl alcohol copolymer in the management of hemoptysis. The indications, immediate control of hemoptysis, and clinical tolerability were analyzed. The indications for embolization were mainly early recurrence of hemoptysis with reperfusion of systemic arteries in seven cases; unstable microcatheter in two cases; large ectopic bronchial artery in two cases; and, in one case each, bronchial arterial bleeding through a small anastomotic network, aneurysm of the left internal thoracic artery in a patient with invasive aspergillosis, a potentially risky connection between the bronchial and right coronary arteries, and occlusion of a systemic artery due to Pryce type 1 intralobar lung sequestration. Hemoptysis was controlled in all but one case and did not recur in the other 14 cases. The injection procedure was well tolerated. Ethylene vinyl alcohol copolymer embolization for hemoptysis of systemic arterial origin is feasible. The agent seems beneficial in the care of patients with recurrent hemoptysis after previous bronchial arterial embolization, patients who would be at high risk during embolization with microparticles, and patients who need embolization of large systemic arteries through small-caliber anastomoses.


A method for maintaining odor-responsive adult rat olfactory receptor neurons in short-term culture.
We report a culture system requiring the addition of freshly made ascorbic acid to the medium, that supports the short-term survival of adult rat olfactory receptor neurons. The cultured neurons exhibit typical voltage-gated currents and are responsive to application of odorants.


An evaluation of temperature stability and resistance in neonatal ventilator circuits.
Gas conditioning minimizes complications associated with invasive ventilation of neonates. Poorly conditioned gas contributes to humidity deficit, facilitates condensate pools, and contributes to safety events. The specific aim was to objectively quantify the temperature drop across the unheated portion of a neonatal circuit and the impact condensation has to resistance to flow in the ventilator circuit. Ventilator circuits and filters were obtained, assembled according to manufacturer recommendations, and operational verification procedures were performed prior to data collection. A neonatal test lung was connected to each Servo-I ventilator with the following settings: pressure control IMV mode; inspiratory pressure: 14 cm H2O to achieve an exhaled tidal volume of 6.0 mL; PEEP: 5 cm H2O; pressure support: 5 cm H2O, FIO2: 0.21; set frequency 40/min; and inspiratory time: 0.4 s. The Fisher and Paykel MR850 and ChonchaTherm Neptune heaters were set at a temperature of 40°C. To evaluate both systems under similar conditions, the ChonchaTherm Neptune heater humidity control was set to midline. Heaters were turned on simultaneously and given 1 h to equilibrate. Readings for room temperature, airway temperature at the patient connection, airway resistance, exhaled tidal volume, and direct observation of circuit condensation and (or) pooling were recorded hourly for a 48-h period. Summary statistics were calculated for the variables of interest. Mean (±SD) air temperature was 26.3°C (±1.4) for the Fisher & Paykel MR850 system and 26.2°C (±1.5), for the ChonchaTherm Neptune system. Mean (±SD) airway resistance was 229.3 cm H2O/L/s (±81.0) for the Fisher & Paykel system and 196.2 cm H2O/L/s (±39.4) for the ChonchaTherm Neptune system. Mean (±SD) tidal volume for the Fisher & Paykel MR850 system was 6.5 mL (±0.4), and for the ChonchaTherm Neptune system was 7.2 mL (±0.6). Circuit condensate increased tidal volume delivery and airway resistance. Temperature at the patient connection was lower than the temperature monitored by the system 12 inches distally, which can negatively impact gas conditioning.