Polyethyleneimine-lipid conjugate-based pH-sensitive micellar carrier for gene delivery.
A low molecular weight polyethyleneimine (PEI 1.8 kDa) was modified with dioleoylphosphatidylethanolamine (PE) to form the PEI-PE conjugate investigated as a transfection vector. The optimized PEI-PE/pDNA complexes at an N/P ratio of 16 had a particle size of 225 nm, a surface charge of +31 mV, and protected the pDNA from the action of DNase I. The PEI-PE conjugate had a critical micelle concentration (CMC) of about 34 μg/ml and exhibited no toxicity compared to a high molecular weight PEI (PEI 25 kDa) as tested with B16-F10 melanoma cells. The B16-F10 cells transfected with PEI-PE/pEGFP complexes showed protein expression levels higher than with PEI-1.8 or PEI-25 vectors. Complexes prepared with YOYO 1-labeled pEGFP confirmed the enhanced delivery of the plasmid with PEI-PE compared to PEI-1.8 and PEI-25. The PEI-PE/pDNA complexes were also mixed with various amounts of micelle-forming material, polyethylene glycol (PEG)-PE to improve biocompatibility. The resulting particles exhibited a neutral surface charge, resistance to salt-induced aggregation, and good transfection activity in the presence of serum in complete media. The use of the low-pH-degradable PEG-hydrazone-PE produced particles with transfection activity sensitive to changes in pH consistent with the relatively acidic tumor environment.


[Genetics of congenital heart diseases].
New insights into the genetics of congenital heart diseases in human beings have been drawn these past years. The identification of genes for heart defects have led to a new clinical approach of these malformations in children and their families. These progresses have been made with the help of positional cloning as well as with the analysis of mouse models. These findings also yielded a new complexity in understanding the development of cardiac defects and led to revise the different classifications for congenital heart defects. Pediatric cardiologists have also improved their efficiency in defining cardiac phenotypes in affected individuals and in pedigrees with recurrent malformations. Genetic heterogeneity has made the molecular approach of a given defect difficult. In addition, intrafamilial variability still has scarce explanations. Finally, the contribution of epigenetic factors has to be kept in mind in specific conditions such as twin gestations. Clinical consequences of these findings remain at the present time limited for the patients themselves but in particular cases, genetic counseling has been dramatically improved.


Muscle fiber diameter assessment in cleft lip using image processing.
To pilot investigation of muscle fiber diameter (MFD) on medial and lateral sides of the cleft in 18 infants with cleft lip with or without cleft palate (CL/P) using image processing. Formalin-fixed paraffin-embedded (FFPE) tissue samples from the medial and lateral sides of the cleft were analyzed for MFD using an image-processing program (ImageJ). For within-case comparison, a paired Student's t test was performed. For comparisons between classes, an unpaired t test was used. Image processing enabled rapid measurement of MFD with majority of fibers showing diameter between 6 and 11 μm. There was no significant difference in mean MFD between the medial and lateral sides, or between CL and CLP. However, we found a significant difference on the medial side (p = .032) between males and females. The image processing on FFPE tissues resulted in easy quantification of MFD with finding of a smaller MFD on the medial side in males suggesting possible differences in orbicularis oris (OO) muscle between the two sexes in CL that warrants replication using larger number of cases. Moreover, this finding can aid subclinical phenotyping and potentially in the restoration of the anatomy and function of the upper lip.


Divergent physical activity and novel alternative responses to high fat feeding in polygenic fat and lean mice.
We determined whether altered physical activity levels might underlie the contrasting adiposity of a divergently selected polygenic murine model of metabolic syndrome (Fat; F) and leanness (Lean; L) mice. We measured physical activity with a long term running wheel experiment and performed an additional high fat diet intervention. Further, we measured posture allocation by visual monitoring within the home cage as a non-exercise correlate of 'normal' physical activity. Whilst initially similar, running wheel activity of the F line declined with age, while the activity of the L line increased. Food intake was higher in the L line and increased with wheel exposure. Vertical rearing measured by video quantification in the home cage, without the stimulus of a running wheel was also significantly higher in the L line. The two lines developed novel alternate strategies to defend their body weight when exposed to high fat diets with a running wheel. F mice increased their running wheel activity, and despite unaltered food intake, still gained weight. L mice reduced their food intake and maintained activity levels without a significant change in body weight. Phenotypic selection for divergence in body fat content has co-segregated with a genetic predisposition for divergent physical activity levels and different strategies for coping with exposure to high fat diets that will facilitate the discovery of the genes underlying these important obesity related traits.


Dihydroorotate dehydrogenase is a high affinity binding protein for A77 1726 and mediator of a range of biological effects of the immunomodulatory compound.
A protein with high affinity (Kd 12 nM) for the immunomodulatory compound A77 1726 has been isolated from mouse spleen and identified as the mitochondrial enzyme dihydroorotate dehydrogenase (EC 188.8.131.52). The purified protein had a pI 9.6-9.8 and a subunit Mr of 43,000. Peptides derived from the mouse protein displayed high microsequence similarity to human and rat dihydroorotate dehydrogenase with, respectively, 35 and 39 out of 43 identified amino acids identical. Dihydroorotate dehydrogenase catalyzes the fourth step in de novo pyrimidine biosynthesis. The in vitro antiproliferative effects of A77 1726 are mediated by enzyme inhibition and can be overcome by addition of exogenous uridine. The rank order of potency of A77 1726 and its analogues in binding or enzyme inhibition was similar to that for inhibition of the mouse delayed type hypersensitivity response. It is proposed that inhibition of dihydroorotate dehydrogenase is an in vivo mechanism of action of the A77 1726 class of compounds. This was confirmed using uridine to counteract inhibition of the murine acute graft versus host response.


Outcomes of Intravascular Ultrasound-Guided Percutaneous Coronary Intervention Among Patients With Acute Coronary Syndrome.
Intravascular ultrasound (IVUS) use in percutaneous coronary intervention (PCI) improves outcomes. However, data on outcomes of IVUS-guided PCI in patients presenting with acute coronary syndrome (ACS) is scarce. Therefore, we sought to study the utilization rate and outcomes of IVUS-guided PCI in patients with ACS. Using the National Readmission database, we identified all patients with ACS who underwent PCI from 2016 to 2019. We used a 1:1 propensity-matched analysis to compare the outcome of patients with ACS who underwent PCI with and without IVUS. In 1,263,997 patients with ACS, 563,521 (44.6%) underwent PCI without IVUS and 40,095 (3.17%) underwent IVUS-guided PCI. A Propensity scored matched comparison of PCI with and without IVUS showed IVUS-guided PCI was associated with a lower risk of in-hospital mortality (odds ratio 0.74, 95% confidence interval 0.64 to 0.85, p <0.01) compared with PCI without IVUS. The utilization of IVUS increased from 2.64% in 2016 to 4.10% in 2019, p <0.001. In conclusion, IVUS-guided PCI is associated with lower in-hospital mortality in patients with ACS, yet the current utilization of IVUS-guided PCI remains low across the United States.


Risk factors for substance use disorders among inpatients with major affective disorders in Taiwan Chinese.
An assessment of potential risk factors for substance use disorders was performed in 49 patients with major affective disorders in a general hospital acute care unit in Taiwan. The major depression patients were noted to have a higher prevalence of substance use disorder than bipolar disorder patients (p = 0.011). Those patients with substance use disorders were noted to be significantly male-predominated (p = 0.043), to have a later onset age of affective disorder (p = 0.009), and to have more visits to the emergency room in the recent one year (p = 0.009). The sedatives-hypnotics-anxiolytics was the most frequently used forms of drug abuse. The major depression patients had a significantly higher sedative-hypnotics-anxiolytics use disorder rate than the bipolar disorder patients (p = 0.001). All patients with alcohol use disorder were noted to have other substance use disorders as well. Fifty six percent of those patients with substance use disorder were polysubstance users. Eighty three percent of the male patients with substance use disorder had poly-substance use disorder.


(E)-4-[2-(4-Eth-oxy-phen-yl)ethen-yl]-1-methyl-pyridinium naphthalene-2-sulfonate.
In the title salt, C16H18NO(+)·C10H7O3S(-), the substituents attached to the central C=C bond adopt a trans conformation and the benzene and pyridinium rings are nearly coplanar, making a dihedral angle of 6.01 (9)°. The crystal structure features weak C-H⋯O hydrogen bonds and C-H⋯π inter-actions .


[Next-generation sequencing].
Next generation or massive parallel sequencing (MPS) is a rapidly advancing method in molecular genetics that will bring significant changes in the personalized medicine field. In this review we briefly describe major types of MPS, including whole-genome, -exome, -transcriptome and amplicon sequencing. We also present an overview of the advantages, drawbacks and possible applications of sequencing technologies available in the Czech Republic.


Fine needle aspiration cytology of neuroblastoma, including peripheral neuroectodermal tumor, with immunocytochemical and ultrastructural confirmation.
Eleven fine needle aspiration (FNA) biopsies were performed on seven children with neuroblastoma, including one patient with a congenital neuroblastoma and another with a peripheral neuroblastoma of the thoracopulmonary region. FNA cytology made the primary diagnosis of neuroblastoma in four of the seven cases. The other biopsies documented local recurrences and metastases to liver, lymph nodes, orbit and breast. The cytologic features included varying numbers of small primitive cells with scanty cytoplasm, poorly to well-formed pseudorosettes, cell processes, a fibrillary matrix and multinucleated ganglion cells. Five of the seven patients had electron microscopic (EM) examination of the FNA specimen, which in all cases confirmed the diagnosis. Batteries of immunoperoxidase stains were performed on all 11 aspirates with variable results. Staining for neuron-specific enolase was positive in four of the five neoplasms tested, although strongly positive in only three of the cases. Staining for neurofilament markers was positive in only two of five tumors. Studies for cytokeratin markers (AE1/3), low-molecular-weight cytokeratin (35BH11), hematopoietic markers (T29/33), immunoglobulin light chains and myoglobin were negative. One case was positive for vimentin. This study attests to the value of FNA cytology in suggesting a correct diagnosis of either primary, recurrent or metastatic neuroblastoma in children. Selective use of immunoperoxidase stains and EM on the aspirates may be of value.


The Controversy of the Most Proper Time for Pregnancy After Bariatric Surgery: a Review of Ten Cases.
Many infertile obese women conceive after bariatric surgery after many years, while guidelines prohibit them from pregnancy for at least 12-18 months after surgery, due to complications. We hereby present 11 cases in our research center, pregnant in less than 12 months after surgery from 2011 to 2015, to evaluate their pregnancy outcome and complications. The mean conception time was 3.6 months after surgery and mean birth weight was 2695 g. Regarding neonatal/fetal complications, there was one case of intrauterine growth retardation and one NICU hospitalization. Among pregnancy complications, eight patients had hyperemesis gravidarum and one had pre-eclampsia. The results of the present study suggest revising setting a contraindication for conception for patients undergoing bariatric surgery.


Phage amplification-based technologies for simultaneous quantification of viable Salmonella in foodstuff and rapid antibiotic susceptibility testing.
Salmonella, especially drug-resistant Salmonella poses a serious threat to food safety and human health. Herein, we proposed a rapid, accurate and sensitive phage amplification-based analysis (PAA) based on an isolated Salmonella phage T156 with broad host range and potent lysis ability for the quantification of viable Salmonella, as well as rapid antibiotic susceptibility testing. This assay has been successful applied to quantify viable Salmonella in food matrices such as milk and lettuce, with a detection limit of 1 colony forming units (CFU) /mL and high specificity for only detect live bacteria not dead bacteria. When combined with real-time PCR (qPCR), PAA-qPCR further reduced the detection time from 6.5 h to 3.5 h, with a detection limit of 10 CFU/mL and without complicate DNA extraction or purification process. Moreover, this assay could also specifically detect drug-resistant Salmonella, shortening the detection time from 24 h to 6.5 h. The results were consistent with that of the conventional paper disc diffusion method (DDM). These PAA methods were evaluated and applied for Salmonella quantification, as well as the detection of drug-resistant Salmonella, providing a promising platform to trace the foodborne pathogen in the complex food matrix, as well as the detection of drug-resistant Salmonella.


Indole-3-carbinol inhibits LPS-induced inflammatory response by blocking TRIF-dependent signaling pathway in macrophages.
Indole-3-carbinol (I3C), a natural hydrolysis product of glucobrassicin, is a member of the Brassica family of vegetables and is known to have various anti-cancer activities. In the present study, we assessed in vitro and in vivo anti-inflammatory effects of I3C and its molecular mechanisms. I3C attenuated the production of pro-inflammatory mediators such as NO, IL-6, and IL-1β in LPS-induced Raw264.7 cells and THP-1 cells through attenuation of the TRIF-dependent signaling pathway. Furthermore, I3C suppressed the infiltration of immune cells into the lung and pro-inflammatory cytokine production such as IL-6, TNF-α in broncho-alveolar lavage fluid (BALF) in the LPS-induced acute lung injury mouse model. I3C also suppressed IL-1β secretion in nigericin treated in vivo model. I3C has potent anti-inflammatory effects through regulating TRIF-dependent signaling pathways, suggesting that I3C may provide a valuable therapeutic strategy in treating various inflammatory diseases.


Proton pump inhibitors and osteoporosis risk: exploring the role of TRPM7 channel.
Long-term use of proton pump inhibitors (PPIs) has been linked to an increased risk of osteoporosis, with various indirect mechanisms so far identified. Although no direct underlying mechanism for effect on bone cells have been investigated with the use of PPIs. Melastatin-like transient receptor potential 7 (TRPM7)channel has been engaged in the proliferation of bone cells. TRPM7 channel is regulated by extracellular Mg2+ and Ca2+ level, that further encourages to analyse if any imbalance with pantoprazole usage could alter bone remodelling process mediated by TRPM7. The present study was conducted to investigate the effect of pantoprazole on the calcium and magnesium level, the cations involved in the bone remodelling process, as well as role of the TRPM7 channel in the proliferation of bone cells. A cytotoxicity study was carried out to study the effect of pantoprazole on the bone cell using MC3T3-E1 cell line, together with the expression of TRPM7 was determined post-pantoprazole treatment. An in vivo study in rats was carried out for estimation of Ca2+, Mg2+ and Ca2+/Mg2+ ratio as well as bone strength was measured over a duration of 4 weeks and 8 weeks with the treatment of pantoprazole. A pilot-scale clinical study was carried out in patients with a fracture to support the evidence of preliminary findings from in-vitro and in vivo studies. MC3T3-E1 cell line treated with pantoprazole showed decreased cell viability in a dose-dependent manner and reduced expression of TRPM7 channel, evidencing interaction of TRPM7 and pantoprazole in the bone remodelling process. A pilot study conducted on 12 patients having major fractures showed changes in serum Mg2+ and Ca2+ levels over a period of 1 month as well as the animal study also showed ionic imbalance over 8-week treatment with pantoprazole. Bone density measured for the patient at the end of the 1-month treatment was found to be in the osteopenic category, together with the animal study which showed a decrease in femur bone strength for the animal treated with pantoprazole over a period of 8 weeks. The study findings proved a negative impact of pantoprazole use on Ca2+ and Mg2+ levels, which can impact TRPM7-mediated bone remodelling which serves to be a possible mechanism for osteoporosis upon pantoprazole use.


B chromosomes reveal a female meiotic drive suppression system in Drosophila melanogaster.
Selfish genetic elements use a myriad of mechanisms to drive their inheritance and ensure their survival into the next generation, often at a fitness cost to its host.1,2 Although the catalog of selfish genetic elements is rapidly growing, our understanding of host drive suppression systems that counteract self-seeking behavior is lacking. Here, we demonstrate that the biased transmission of the non-essential, non-driving B chromosomes in Drosophila melanogaster can be achieved in a specific genetic background. Combining a null mutant of matrimony, a gene that encodes a female-specific meiotic regulator of Polo kinase,3,4 with the TM3 balancer chromosome creates a driving genotype that is permissive for the biased transmission of the B chromosomes. This drive is female-specific, and both genetic components are necessary, but not individually sufficient, for permitting a strong drive of the B chromosomes. Examination of metaphase I oocytes reveals that B chromosome localization within the DNA mass is mostly abnormal when drive is the strongest, indicating a failure of the mechanism(s) responsible for the proper distribution of B chromosomes. We propose that some proteins important for proper chromosome segregation during meiosis, like Matrimony, may have an essential role as part of a meiotic drive suppression system that modulates chromosome segregation to prevent genetic elements from exploiting the inherent asymmetry of female meiosis.


The Elimination of Lymphatic Filariasis: A Strategy for Poverty Alleviation and Sustainable Development - Perspectives from the Philippines.
BACKGROUND: Within the Philippines areas endemic for lymphatic filariasis are in regions with the highest incidence of poverty. Out of a total of 79 provinces, 39 have a higher poverty incidence than the national average and 30 of these 39 provinces are endemic for lymphatic filariasis. DISCUSSION: Recognizing that provinces endemic for lymphatic filariasis (LF) are also the poorest provinces, the elimination of lymphatic filariasis in these areas presents significant opportunities to reduce poverty and inequalities in health. The implementation of an effective national programme for the elimination of lymphatic filariasis will provide means for sustainable development at national, local and community levels. SUMMARY: The elimination of lymphatic filariasis as a public health problem is a 20-year strategic plan for the world community, with the vision of all endemic communities free of transmission of lymphatic filariasis by 2020 and with the commitment to ensure the delivery of quality technologies and human services to eliminate lymphatic filariasis worldwide through a multi-stakeholder global alliance of all endemic countries. This global goal of elimination of lymphatic filariasis is a significant opportunity for partnerships - a world with less poverty through sustainable development and free from the scourge of lymphatic filariasis.


Simplified ontologies allowing comparison of developmental mammalian gene expression.
Model organisms represent an important resource for understanding the fundamental aspects of mammalian biology. Mapping of biological phenomena between model organisms is complex and if it is to be meaningful, a simplified representation can be a powerful means for comparison. The Developmental eVOC ontologies presented here are simplified orthogonal ontologies describing the temporal and spatial distribution of developmental human and mouse anatomy. We demonstrate the ontologies by identifying genes showing a bias for developmental brain expression in human and mouse.


Chronic in vivo ethanol administration alters the sensitivity of adenylate cyclase coupling in homogenates of rat brain.
The high activities of adenylate cyclase, phosphodiesterase and protein kinases in the synaptic terminals of the central nervous system makes these enzymes prime candidates for the in vivo actions of ethanol. Adult female rats were fed a liquid diet containing ethanol as 35% of the available calories for 6 days. This resulted in a decrease (22-45%) in the basal activity of adenylate cyclase, as determined by cyclic 3',5'-adenosine monophosphate (cAMP) production, in homogenates of all brain areas tested. In these homogenates the ability of guanosine triphosphate and noradrenaline to stimulate basal cyclase activity was severely reduced. These results suggest that ethanol administration causes an uncoupling of the beta-receptor/adenylate cyclase cascade and an interruption of the control of the synthesis of cAMP.


MAPK/ERK-dependent translation factor hyperactivation and dysregulated laminin γ2 expression in oral dysplasia and squamous cell carcinoma.
Lesions displaying a variety of dysplastic changes precede invasive oral and epidermal squamous cell carcinoma (SCC); however, there are no histopathological criteria for either confirming or staging premalignancy. SCCs and dysplasias frequently contain cells that abnormally express the γ2 subunit of laminin-332. We developed cell culture models to investigate γ2 dysregulation. Normal human keratinocytes displayed density-dependent repression of γ2, whereas premalignant keratinocytes and SCC cells overexpressed γ2 and secreted laminin assembly intermediates. Neoplastic cells had hyperactive EGFR/MAPK(ERK) signaling coordinate with overexpressed γ2, and EGFR and MEK inhibitors normalized γ2 expression. Keratinocytes engineered to express HPV16 E6 or activated mutant HRAS, cRAF1, or MEK1 lost density repression of γ2 and shared with neoplastic cells signaling abnormalities downstream of ERK, including increased phosphorylation of S6 and eIF4 translation factors. Notably, qPCR results revealed that γ2 overexpression was not accompanied by increased γ2 mRNA levels, consistent with ERK-dependent, eIF4B-mediated translation initiation of the stem-looped, 5'-untranslated region of γ2 mRNA in neoplastic cells. Inhibitors of MEK, but not of TORC1/2, blocked S6 and eIF4B phosphorylation and γ2 overexpression. Immunostaining of oral dysplasias identified γ2 overexpression occurring within fields of basal cells that had elevated p-S6 levels. These results reveal a causal relationship between ERK-dependent translation factor activation and laminin γ2 dysregulation and identify new markers of preinvasive neoplastic change during progression to SCC.


Superimposed spontaneous esophageal perforation in congenital esophageal stenosis.
Congenital esophageal stenosis (CES) is a very rare embryologic anomaly of tracheoesophageal development that requires lifelong management, usually with frequent esophageal dilations and dietary precautions. We present a patient with spontaneous thoracic esopahgeal perforation in a setting of CES. The patient, a 27-year-old male, recovered uneventfully following open primary esophageal closure. A brief but comprehensive discussion of CES follows the case report.


A Patient Presents with Longstanding, Severe LV Dysfunction. Is There a Role for Additional Risk Stratification Before ICD?
Increased longevity of patients with systolic heart failure caused by the use of implantable cardioverter-defibrillators (ICDs) is one of the most successful achievements in contemporary medicine. During the last 2 decades, the scientific community has striven to increase the benefits of ICD usage by specifying indications for primary prevention ICD implantation. Left ventricular ejection fraction is neither highly specific nor is it a highly sensitive risk marker of sudden cardiac death. The authors discuss risk-stratification approaches in different patient populations with structural heart disease and systolic heart failure.


The accidental conservationist: William T. Hornaday, the Smithsonian bison expeditions and the US National Zoo.
In 1886 William T. Hornaday, the Chief Taxidermist of the US National Museum, led the "Smithsonian Institution Buffalo Outfit" to Montana. The American bison was fast heading toward extinction and the mission of the expedition was to locate those of the elusive animals that remained and obtain specimens for the scientific collection at the museum. The successful expedition produced the most complete scientific series, and the finest artistic grouping of taxidermied specimens of the American bison anywhere in the world. Haunted by the scattered skeletal remains of the millions of slaughtered bison that peppered the Eastern Montana Plains, Hornaday fought to establish the National Zoological Park, which would provide sanctuary for bison and other endangered species, and a captive breeding program, which would result in the eventual reintroduction of the American bison to the wild.


Temporal and geographical clustering of Kawasaki disease in Japan: 2007-2012.
Since 1987, no study has reported the municipal-level geographical clustering of Kawasaki disease (KD) in Japan. Therefore, the aim of the present study was to identify the temporal and municipal-level geographical clustering of KD. The annual incidence rates of KD for each municipality were calculated using nationwide data from 73 758 patients with KD (2007-2012). To determine whether temporal and municipal-level clustering existed, we calculated the correlations of the annual incidence rates for each municipality during the study years, and compared these rates with those of the adjacent municipalities. Spatial scanning analysis was used to identify the geographical clusters for each year, and the incidence rates in those clusters were compared with the rates in the surrounding region. The annual national incidence rate of KD, adjusted for the prefecture-specific response rate, was 322.45 patients per 100 000 children aged 0-4 years. The correlation between the annual incidence rates during 2 consecutive years was significantly positive (coefficients, 0.149-0.428). On spatial scanning analysis, the most likely clusters were in the Tokyo metropolitan area during 2007-2010 and 2012, and in Kumamoto prefecture during 2011. Kawasaki disease exhibits temporal and municipal-level clustering.


Custodians of Information: Patient and Physician Views on Sharing Medical Records in the Acute Care Setting.
In the UK, in the acute in-patient setting, the only information that a patient receives about their medical care is verbal; there is no routine patient access to any part of the medical record. It has been suggested that this should change, so that patients can have real-time access to their notes, but no one has previously explored patient or clinician views on the impact this might have. Semi-structured interviews were conducted with 12 patients and 13 doctors about their experience of information sharing in the context of the acute care setting, and their views on sharing all of the medical records, or a summary note. Interviews were transcribed verbatim, double coded and analyzed using the constant comparative method. Patients were not given written information and did not ask questions even when they wanted to know things. Patients and doctors supported increased sharing of written information, but the purpose of the medical record - and the risks and benefits of sharing it - were disputed. Concerns included disclosing uncertainty, changing what was written, and causing patient anxiety. Benefits included increased transparency. Use of a summary record was welcomed as a way to empower patients, while doctors felt they had a responsibility to curate what information was given and when. A clinical summary for patients would be of benefit to doctors, nurses, patients and their relatives. It should be designed to reflect the needs of all users, and evaluated to consider patient-relevant outcomes and resource implications.


Left atrial dysfunction is a predictor of postcoronary artery bypass atrial fibrillation: association of left atrial strain and strain rate assessed by speckle tracking.
Even though atrial fibrillation (AF) is the most common arrhythmia after coronary artery bypass grafting (CABG), its etiology remains poorly understood. Several factors are linked to postoperative AF (POAF), including advanced age and systemic inflammation. However, left atrial (LA) contractile dysfunction has not been evaluated in the perioperative scenario. To evaluate LA function through strain and strain rate in patients with coronary artery disease undergoing CABG and its correlation with POAF. We studied 70 patients undergoing CABG in sinus rhythm at the time of surgery. Preoperative echocardiography with evaluation of LA strain and strain rate by speckle tracking was performed. The occurrence of POAF was evaluated by continuous monitoring. Baseline and postoperative C-reactive protein (CRP) levels were measured to evaluate systemic inflammation. After 1-week follow-up 26% of subjects developed AF. LA strain s wave (LASs) and LA strain rate s (LASRs) and a wave (LASRa) were significantly decreased in patients who developed POAF: LASs (10 ± 1% vs. 24 ± 1%, P < 0.001), LASRs (0.6 ± 0.1 sec(-1) vs. 1.2 ± 0.1 sec(-1) , P < 0.001), LASRa (-0.6 ± 0.1 sec(-1) vs. -1.8 ± 0.1 sec(-1) , P < 0.001). LASRs, LASRa, age, and LA volume were independent predictors of POAF. CRP at baseline was similar irrespective of POAF development. LA dysfunction, evaluated by strain and strain rate is an independent predictor of POAF and contributes to classic risk factors like age and atrial volume.


Addressing Weight Suppression to Improve Treatment Outcome for Bulimia Nervosa.
Cognitive behavioral therapy (CBT) is regarded as the gold-standard treatment for bulimia nervosa (BN), yet despite impressive empirical support for its effectiveness, over 50% of patients fail to achieve abstinence from binge eating and purging by the end of treatment. One factor that may contribute to reduced efficacy rates in CBT is weight suppression (WS; the difference between a person's highest weight ever at their adult height and current weight). A growing body of research indicates that WS in patients with BN may have a clinically significant effect on symptomatology and prognosis. However, the current cognitive behavioral framework for BN does not explicitly acknowledge the role of WS in the onset or maintenance of BN symptoms and does not provide guidance for clinicians on how to address WS during treatment. The relationship between WS, biological pressure to regain lost weight, and the maintenance of BN symptoms suggest that current cognitive behavioral models of BN may be improved by considering the role of WS and exploring needed treatment modifications. Indeed, a reconceptualization of existing models may offer insight into potential strategies that can be used to reduce the susceptibility to treatment dropout, nonresponse, and relapse. It is therefore necessary to consider whether, and how, clinicians' consideration of WS during case conceptualization and treatment planning could serve to improve CBT outcomes. The current review explores ways in which high WS could contribute to poor CBT outcomes, provides preliminary clinical recommendations for incorporating WS into existing cognitive behavioral treatments based on extant data and clinical wisdom, and proposes suggestions for future research needed in this domain.


Potential use of a canine whole blood culture system to evaluate the immune response to Leptospira.
In susceptible hosts, protection from Leptospira infection is mediated by the innate immune response at the point of entry and humoral immunity. Thus, identifying and segregating the initial host response at the representative host-pathogen interface is needed to understand the typical outcomes of Leptospira infection, clearance, persistence, or disease. An in vitro whole blood culture system to study the overall immune response using pathogenic and non-pathogenic Leptospira strains was explored in this study. Using an ELISA, increased IL-8, TNF alpha, and IL-1 in blood samples stimulated with pathogenic and nonpathogenic Leptospira compared to unstimulated controls were detected. In RT2 Profiler PCR Array assays, consistent upregulation of 22 genes and downregulation of 25 genes were observed. Few of the notable upregulated genes included BPI, CCL3, CXCL2, IL-6, IL-8, TLR1, TLR2, TLR6, and TNF and downregulated genes included, LBP, LYZ, MPO, MYD88. IFNβ was upregulated in samples treated with pathogenic Leptospira and IL-1β was upregulated in samples treated with nonpathogenic Leptospira. Toll- like Receptor signaling and expression of pattern recognition receptors were two of the five prominent canonical pathways observed. Individual deconvolution of each of the specific and significant pathways observed in this study may improve the understanding of the pathogenesis of this important zoonotic agent. The use of this system in conjunction with whole transcriptome analysis in a larger population, may unveil the robust nature of host/Leptospira interaction.


Outcomes after ORIF of Bicondylar Schatzker VI (AO type C) Tibial Plateau Fractures in an Elderly Population.
The surgical management of bicondylar tibial plateau (BTP) fractures in elderly patients aims to restore knee stability while minimizing soft tissue complications. The purpose of this study was to compare injury characteristics and surgical outcomes after ORIF of BTP fractures (AO/OTA 41-C (Schatzker VI)) in young (< 50 years) versus elderly (> 65 years) patients. A retrospective cohort study was conducted using data from two American College of Surgeons (ACS) level I trauma centers. Inclusion criteria were: (1) age 18 years or older, (2) bicondylar tibial plateau fracture (AO/OTA 41-C or Schatzker VI), (3) treatment with ORIF, and (4) minimum of 6 months follow-up. Patients between 50 and 65 years of age were excluded. Data collection was performed by reviewing electronic medical records, operative reports, and radiology reports. We identified 323 patients (61% male) with 327 BTP fractures and a median follow-up of 685 days. There were 230 young patients (71%) < 50 years and 93 elderly patients (29%) >6 5 years at time of presentation. Elderly patients were significantly more likely to have a low energy mechanism of injury (44.6 vs. 16.2%, p < 0.001), and present with diabetes (19.4 vs. 4.4%, p < 0.001) or coronary artery disease (12.9 vs. 1.3%, p < 0.001). Elderly patients were also significantly less likely to undergo staged management with initial knee-spanning external fixation followed by delayed ORIF (19.2 vs. 33.9%, p = 0.008). Elderly patients had a lower arc of motion at final follow-up (105 vs. 113°, p < 0.001) and reduced PROMIS-10 function scores (43.8 vs. 49.8, p = 0.013). No differences were observed in rates of superficial infection, deep infection, reoperation, or EQ-5D scores between age groups. This is the largest study to compare injury characteristics and outcomes after ORIF of BTP fractures according to age. Elderly patients (age > 65 years) sustained BTP fractures by lower energy mechanisms than their younger counterparts with similar fracture patterns and were often managed with ORIF. The results of this study suggest that ORIF of BTP fractures in elderly patients is associated with similar complication rates and outcomes as in younger patients despite higher comorbidities and poorer bone quality in the elderly population.


Safety and efficacy of Prunella vulgaris preparation in adjuvant treatment of thyroid nodules: A meta-analysis.
Prunella vulgaris (PV), a traditional Chinese medical herb, is considered beneficial for some thyroid diseases. However, the effectiveness is not consistent in different studies. This review compiles the evidence from randomized controlled trials (RCTs) and quantifies the effects of PV preparation on thyroid nodules. Eight databases were searched up to April 2021 to identify eligible studies. Only RCTs were included. Meta-analysis of homogeneous studies was performed by RevMan5.3 software. Cochrane risk of bias assessment tool version 2.0 was used to assess the risk of bias of each trial. The research screening, data extraction, and risk of bias assessment were employed by 2 reviewers independently, and disagreement will be decided by a third senior reviewer. The risk ratio (RR), mean difference (MD) and corresponding 95% confidence interval (CI) of each study are summarized. Thirteen RCTs with 1468 patients were included in this study. A meta-analysis showed that the RR of the clinical efficacy of PV combined with levothyroxine sodium tablets was 1.22 (95% CI [1.11, 1.33]). The MD of thyroid nodule diameter was -0.43 (95% CI [-0.63, -0.22]). The MD of free triiodothyronine and free tetraiodothyronine levels was -1.99 (95% CI [-3.14, -0.86]) and -3.20 (95% CI [-5.50, -0.89]), respectively. The RR of the adverse reaction rate was 0.67 (95% CI [0.36, 1.22]), and the RR of the clinical efficacy of PV preparation combined with thyroxin tablets was 1.29 (95% CI [1.03, 1.62]). PV combined with levothyroxine sodium tablets or thyroxin tablets has more benefits for thyroid nodules, further improving the clinical efficiency, reducing the diameter of nodules and reducing the occurrence of adverse reactions. However, the quality of these studies is uncertain, and higher quality and more RCTs are needed to provide comprehensive evidence-based medical evidence in the future.


Cellular microRNA-127-3p suppresses oncogenic herpesvirus-induced transformation and tumorigenesis via down-regulation of SKP2.
Kaposi's sarcoma-associated herpesvirus (KSHV) causes the endothelial tumor KS, a leading cause of morbidity and mortality in sub-Saharan Africa. KSHV-encoded microRNAs (miRNAs) are known to play an important role in viral oncogenesis; however, the role of host miRNAs in KS tumorigenesis remains largely unknown. Here, high-throughput small-RNA sequencing of the cellular transcriptome in a KS xenograft model revealed miR-127-3p as one of the most significantly down-regulated miRNAs, which we validated in KS patient tissues. We show that restoration of miR-127-3p suppresses KSHV-driven cellular transformation and proliferation and induces G1 cell cycle arrest by directly targeting the oncogene SKP2. This miR-127-3p-induced G1 arrest is rescued by disrupting the miR-127-3p target site in SKP2 messenger RNA (mRNA) using gene editing. Mechanistically, miR-127-3p-mediated SKP2 repression elevates cyclin-dependent kinase (CDK) inhibitor p21Cip1 and down-regulates cyclin E, cyclin A, and CDK2, leading to activation of the RB protein tumor suppressor pathway and suppression of the transcriptional activities of E2F and Myc, key oncoprotein transcription factors crucial for KSHV tumorigenesis. Consequently, metabolomics analysis during miR-127-3p-induced cell cycle arrest revealed significant depletion of dNTP pools, consistent with RB-mediated repression of key dNTP biosynthesis enzymes. Furthermore, miR-127-3p reconstitution in a KS xenograft mouse model suppresses KSHV-positive tumor growth by targeting SKP2 in vivo. These findings identify a previously unrecognized tumor suppressor function for miR-127-3p in KS and demonstrate that the miR-127-3p/SKP2 axis is a viable therapeutic strategy for KS.


Simultaneous multiplane imaging with a distorted diffraction grating.
We describe a simple technique for simultaneously imaging multiple layers within an object field onto a single camera. The approach uses a binary diffraction grating in which the lines are distorted such that a different level of defocus is associated with each diffraction order. The design of the gratings is discussed, and their ability to image multiple object planes is validated experimentally. Extension of the technique for spherical-aberration correction is described, and it is shown how the gratings can be used as part of a wave-front-sensing system.


Assessment of a flow cytometry technique for studying signaling pathways in platelets: Monitoring of VASP phosphorylation in clinical samples.
A recently released kit (PerFix EXPOSE) was reported to improve the measurement of the degree of phosphorylation of proteins in leukocytes by flow cytometry. We tested its adaptation for platelets to monitor vasodilator-stimulated phosphoprotein (VASP) phosphorylation, which is the basis of a currently used test for the assessment of the pharmacological response to P2Y12 antagonists (PLT VASP/P2Y12). The PerFix EXPOSE kit was compared to the PLT VASP/P2Y12 kit by using blood samples drawn at 24 h post clopidogrel dose from 19 patients hospitalized for a non-cardio-embolic ischemic stroke and treated with clopidogrel monotherapy for at least five days in an observational study. The platelet PerFix method was based on adaptation of the volume of the sample, the centrifugation speed and the incubation temperature. Poor agreement between prevention by adenosine diphosphate (ADP) of PGE1-induced cAMP-mediated VASP phosphorylation and ADP induced aggregation assessed by Light Transmittance Aggregometry was found. We found a significant correlation between the PLT VASP/P2Y12 kit and the PerFix EXPOSE kit. The PerFix EXPOSE kit may also be helpful to monitor adverse effects of second-generation tyrosine kinase inhibitors on platelets.


Pulmonary T2N0 adenocarcinoma with metastasis to a lymph node in the thoracic wall.
We experienced a rare case of lung cancer without hilar/mediastinal nodal involvement or direct invasion to the thoracic wall, but with metastasis to a lymph node in the thoracic wall. A 72-year-old woman with lung cancer was admitted to our hospital for the surgical therapy. She had suffered from right pleuritis in her childhood. During the dissection of the pleural adhesion around the whole lung, one small black lymph node was found in the thoracic wall and resected. Then, right middle and lower lobectomy and systematic nodal dissection were performed. The postoperative pathological examination revealed that nodal involvement was not observed in all samples except in the lymph node in the thoracic wall. In lung cancer patients with broad pleural adhesion, we should pay attention to lymph nodes in the thoracic wall. If we find them, the nodes should be resected for accurate staging.


The circadian clock, aging and its implications in cancer.
Circadian clock orchestrates the intergenic biochemical, physiological and behavioral changes to form an approximate 24h oscillation through the transcription-translation feedback loop (TTFL). Mechanistically, a heterodimer of transcriptional activator formed by BMAL1 and CLOCK, governs the expression of its transcriptional repressors, CRY, PER and REV-ERBα/β proteins, thereby controlling more than 50 % of protein encoding genes in human. There is also increasing evidence showing that tumor microenvironment can disrupt specific clock gene functions to facilitate tumorigenesis. Although there is great progress in understanding the molecular mechanisms of the circadian clock, aging and cancer, elucidating their complex relationships among these processes remains challenging. Herein, the optimization of the chronochemotherapy regimen has not been justified yet for treatment of cancer. Here, we discuss the hypothesis of relocalization of chromatin modifiers (RCM) along with function(s) of the circadian rhythm on aging and carcinogenesis. We will also introduce the function of the chromatin remodeling as a new avenue for rejuvenation of competent tissues to combat aging and cancer.


Comparison of a non-competitive two-site RIA and an inhibition RIA for the detection of myelin basic protein in cerebrospinal fluid.
Two radioimmunoassay (RIA) procedures were used to measure human myelin basic protein (HBP) in cerebrospinal fluid (CSF): (1) an inhibition RIA, with the use of TNP-conjugated anti-BP IgG, 125I-labelled HBP, and anti-TNP-coated polystyrene beads, and (2) a non-competitive two-site RIA, with the use of Sepharose-coupled anti-BP antibodies and 125I-labeled anti-BP IgG. The two-site RIA detects less HBP in CSF than the inhibition RIA, partly due to the presence of HBP fragments in CSF that are detected by the inhibition assay, but less by the two-site RIA. The correlation was improved when in the two-site RIA Sepharose-coupled anti-BP antibodies were changed. Because certain substances (such as autoantibodies to HBP) may give false-positive results in the competitive RIA but not in the two-site RIA, we conclude that a combination of the (more sensitive) inhibition RIA with the (more specific) two-site assay provides a more reliable HBP assay than either assay alone.


Submissive behavior in mice as a test for antidepressant drug activity.
Previously, with the administration of antidepressant drugs, it has been demonstrated that the rat model of clinical depression, known as the reduction of submissive behavior model (RSBM), has considerable validity. The present study is an attempt to extend the model to mice. Several antidepressant drugs as well as a number of non-antidepressant agents were administered to mice that had been identified as submissive in a behavioral testing situation. Imipramine, desipramine, amoxapine and fluoxetine, representing three different classes of antidepressant drugs, were each able to increase competitive behavior in submissive mice and to decrease the dominance level between dominant and submissive mice in the behavioral tests. The stimulant amphetamine also reduced submissive behavior while yohimbine (also a stimulant), and the antianxiety agent diazepam had no such effect. The neuroleptic drug thiothixen had antidepressant-like effect on submissive C57BL/6J mice behavior. We conclude that like the rat model of depression from which it was developed, the mouse model responds to various antidepressants as predicted and thus may serve as a potential model of clinical depression.


Gamma Attenuation Coefficients of Nano Cadmium Oxide/High density Polyethylene Composites.
In the present work, high density polyethylene (HDPE) matrix mixed with micro-sized and nano-sized Cadmium oxide (CdO) particles of different concentrations were prepared by compression molding technique. The aim of the study is to investigate the effect of particle size and weight percentage of CdO particles on the gamma radiation shielding ability of CdO/HDPE composites. The mass attenuation coefficients of pure HDPE, micro-CdO/HDPE and nano-CdO/HDPE composites were evaluated at photon energies ranging from 59.53 keV to 1408.01 keV using standard radioactive point sources [241Am, 133Ba, 137Cs, 60Co and 152Eu]. Adding micro and nano CdO particles to the HDPE matrix clearly increases the mass attenuation coefficients of the composites and the improvement is more significant at low γ-ray energies. The effect of particle size of CdO filler has an important role on the shielding ability of the composite. The experimental results reveal that, the composites filled with nano-CdO have better γ-radiation shielding ability compared to that filled with micro-CdO at the same weight fraction. A relative increase rate of about 16% is obtained with nano-CdO content of 40 wt% at 59.53 keV, which attributed to the higher probability of interaction between γ-rays and nanoparticles. From this study, it can be concluded that nano-CdO has a good performance shielding characteristic than micro-CdO in HDPE based radiation shielding material.


Age dependency of ABO histo-blood group antibodies: reexamination of an old dogma.
Current textbooks for transfusion medicine state that anti-A and/or anti-B (anti-A/B) agglutination titers--and thus the respective antibody concentrations--reach their maximum in individuals 5 to 10 years old and then gradually decline with the increasing age of the individual. This statement is largely based on a study by Thomsen and Kettel that dates to 1929. In the present article, ABO antibodies in sera of 175 healthy persons aged 61 to 97 years, as well as sera of 170 newborn infants and children aged 0 to 17 years, were analyzed. Microhemagglutination tests were performed with all sera and complemented by ABO enzyme-linked immunosorbent assays to measure the immunoglobulin class (IgM, IgG, and IgA) of the anti-A/B. As in a previous study using sera of persons aged 20 to 67 years, individual differences exceeded age-related changes for all variables. Median values of IgG and IgA anti-A/B were elevated in elderly persons of blood group O, whereas no significant changes were observed in other variables. In particular, the decrease in agglutination titers with the increasing age of the individuals was far less pronounced than previously described; even in sera of persons aged 90 to 97 years, median agglutination titers of 128 were found. Results in the sera of children confirm previously reported data that agglutination titers and IgM anti-A/B reached adult levels at the age of 5 to 10 years.


Multifunctional nanoadditives for the thermodynamic and kinetic stabilization of enzymes.
Stabilization of enzymes has become a major focus in the quest to improve the activity, sustainability and recyclability of enzymes for their successful integration into both industry and medicine. Here, we describe the kinetic and thermodynamic stabilization of a variety of enzymes in the presence of cationic multifunctional polymeric nanoparticles.


Supervised machine learning for diagnostic classification from large-scale neuroimaging datasets.
There are growing concerns about the generalizability of machine learning classifiers in neuroimaging. In order to evaluate this aspect across relatively large heterogeneous populations, we investigated four disorders: Autism spectrum disorder (N = 988), Attention deficit hyperactivity disorder (N = 930), Post-traumatic stress disorder (N = 87) and Alzheimer's disease (N = 132). We applied 18 different machine learning classifiers (based on diverse principles) wherein the training/validation and the hold-out test data belonged to samples with the same diagnosis but differing in either the age range or the acquisition site. Our results indicate that overfitting can be a huge problem in heterogeneous datasets, especially with fewer samples, leading to inflated measures of accuracy that fail to generalize well to the general clinical population. Further, different classifiers tended to perform well on different datasets. In order to address this, we propose a consensus-classifier by combining the predictive power of all 18 classifiers. The consensus-classifier was less sensitive to unmatched training/validation and holdout test data. Finally, we combined feature importance scores obtained from all classifiers to infer the discriminative ability of connectivity features. The functional connectivity patterns thus identified were robust to the classification algorithm used, age and acquisition site differences, and had diagnostic predictive ability in addition to univariate statistically significant group differences between the groups. A MATLAB toolbox called Machine Learning in NeuroImaging (MALINI), which implements all the 18 different classifiers along with the consensus classifier is available from Lanka et al. (2019) The toolbox can also be found at the following URL: https://github.com/pradlanka/malini .


Brain Surgery for Medically Intractable Epilepsy.
This review covers the broad topic of brain surgery in the treatment of pediatric intractable epilepsy. The authors review the latest advancements in the presurgical workup as well as the mandatory tests needed to explore the epilepsy workup in these children. They describe the different types of epilepsy from a surgical standpoint (temporal, extratemporal, multifocal, and hemispheric epilepsies) and various surgical procedures that can be proposed depending on the clinical scenario: lesionectomies, lobectomies, hemispherectomies, neuromodulation, and palliative surgeries. They also describe the key differences of the pediatric patient as compared with the adult patient in such pathologic conditions.


Inter-hemispheric functional and anatomical connectivity abnormalities in traffic accident-induced PTSD: a study combining fMRI and DTI.
Aberrant brain functional and structural changes are considered to be one of the important mechanisms underlying post-traumatic stress disorder (PTSD). However, it remains unclear whether inter-hemispheric connection is changed. The current study aimed to identify the inter-hemispheric functional and anatomical connectivity changes in patients who consequently develop PTSD using the voxel-mirrored homotopic connectivity (VMHC) analysis and diffusion tractography techniques. Resting-state fMRI and DTI data were acquired on victims who had experienced traffic accidents within 2 days after the traumatic event. The diagnosis was made using the Clinician-Administered PTSD Scale at 1 or 6 months later. Fifteen trauma-exposed victims met the criteria for diagnosis of PTSD and 14 trauma-exposed victims who did not develop PTSD at 6 months after trauma were selected as the control group. Compared with the victims without PTSD, the victims with PTSD exhibited an abnormal homotopic pattern with decreased VMHC in the superior/middle frontal gyrus before diagnosis. The regions showing abnormal functional connectivity were then chosen as regions of interest for an analysis of DTI tractography. Decreased fractional anisotropy values in the genu of the corpus callosum were found in the victims with PTSD. Greater WM disruptions within 2 days predicted greater symptom severity at diagnosis. The study was lack of comparison with controls who did not experience a traumatic event. Our results suggest that the inter-hemispheric functional and structural connectivity is impaired in PTSD within 2 days, which may be the potential marker showing predisposition towards developing PTSD.


Sildenafil treatment prevents glomerular hypertension and hyperfiltration in rats with renal ablation.
Sildenafil treatment ameliorates progressive renal injury resulting from extensive renal ablation; however, modifications induced by sildenafil in the glomerular hemodynamic pathophysiology of the remnant kidney have not been investigated. To determine the effects of sildenafil in the glomerular microcirculation and their relation to histological damage in the renal ablation model. Micropuncture studies were performed 60 days after 5/6 nephrectomy in rats that received no treatment, sildenafil (5 mg/kg/day) and reserpine, hydralazine and hydrochlorothiazide to maintain the blood pressure within normal levels. Sham-operated rats untreated and treated with sildenafil served as controls. As expected, renal ablation induced systemic and glomerular hypertension, hyperfiltration, proteinuria, glomerulosclerosis and tubulointerstitial inflammatory damage in the remnant kidney. Sildenafil treatment prevented single-nephron hyperfiltration and hypertension, suppressed renal arteriolar remodeling, ameliorated systemic hypertension and proteinuria, increased urinary excretion of cGMP and NO(2)(-)/NO(3)(-), decreased oxidative stress and improved histological damage in the remnant kidney. Normalization blood pressure with reserpine, hydralazine and hydrochlorothiazide did not modify glomerular hemodynamics, proteinuria or histological changes induced by renal ablation. Beneficial effects of sildenafil in the remnant kidney are associated with a reduction in the arteriolar remodeling, renal inflammatory changes and prevention of changes in the glomerular microcirculation.


ICP-OES and Micronucleus Test to Evaluate Heavy Metal Contamination in Commercially Available Brazilian Herbal Teas.
Increased tea consumption in combination with intensive pesticide use is generating heavy metal contaminations amongst Brazilian tea consumers, causing health concerns. Inductively coupled plasma optical emission spectrometry (ICP-OES) was applied to quantify minerals and heavy metals such as aluminum, barium, cadmium, lead, cobalt, copper, chromium, tin, manganese, molybdenum, nickel, selenium, silver, thallium, vanadium and zinc in Brazilian chamomile, lemongrass, fennel and yerba mate teas. Teas, purchased in local supermarkets, were prepared using infusion and acid digestion. Higher concentrations of Al were present in all samples. In the digested samples, the Al mean concentration was 2.41 μg g(-1) (sd = 0.72) for fennel and 33.42 μg g(-1) (sd = 17.18) for chamomile, whilst the sample C for chamomile tea presented the highest concentration with 51.62 μg g(-1) (sd = 9.17). The safety relation in decreasing order is fennel, lemongrass, chamomile and yerba mate. Chemometric analyses demonstrated a strong correlation between the elements Cd and Pb in the samples. Yerba mate had the highest amount of metal (100 mg kg(-1)), being the subject of a micronucleus test assay for cytotoxicity. The metals found in Yerba mate did not present cytotoxicity/mutagenicity using the micronucleus test. The inorganic contaminants in teas should have their impact carefully monitored.


DeepASmRNA: Reference-free prediction of alternative splicing events with a scalable and interpretable deep learning model.
Alternative splicing is crucial for a wide range of biological processes. However, limited by the availability of reference genomes, genome-wide patterns of alternative splicing remain unknown in most nonmodel organisms. We present an attention-based convolutional neural network model, DeepASmRNA, for predicting alternative splicing events using only transcriptomic data. DeepASmRNA consists of two parts: identification of alternatively spliced transcripts and classification of alternative splicing events, which outperformed the state-of-the-art method, AStrap, and other deep learning models. Then, we utilize transfer learning to increase the performance in species with limited training data and use an interpretation method to decipher splicing codes. Finally, applying Amborella, DeepASmRNA can identify more AS events than AStrap while maintaining the same level of precision, suggesting that DeepASmRNA has superior sensitivity to identify alternative splicing events. In summary, DeepASmRNA is scalable and interpretable for detecting genome-wide patterns of alternative splicing in species without a reference genome.


In vitro pepsin resistance of proteins: Effect of non-reduced SDS-PAGE analysis on fragment observation.
The introduction of novel proteins to food products carries with it the need to assess the potential allergenicity of such materials. Resistance to in vitro pepsin digestion is one parameter considered in such a risk assessment using a weight of evidence approach; however, the methodology used to investigate this has not been fully standardised. In vitro pepsin resistance assays typically involve SDS-PAGE performed under reducing conditions, with limited published data available on the assay using non-reducing conditions despite the need to consider non-reducing analysis techniques having been highlighted by regulatory bodies such as the European Food Safety Authority (EFSA). The purpose of the work reported here was to investigate the applicability of (and additional insight provided by) non-reducing analyses, by digesting a set of proteins using a ring-trial validated method, with analysis by SDS-PAGE under both reducing and non-reducing conditions. In silico prediction of digest fragments was also investigated. Significant differences were observed between results under reduced and non-reduced conditions for proteins in which disulphide bonds have a major role in protein structure, such as ribulose 1,5-diphosphate carboxylase (RUBISCO) and bovine serum albumin. For proteins with no or few disulphide bonds, no significant differences were seen in the results. Structural information such as disulphide bond numbers and positions should be considered during experimental design, as a non-reduced approach may be appropriate for some proteins. The in silico approach was a useful tool to suggest potential digest fragments, however the predictions were not always confirmed in vitro and should be considered a guide only.


Intralipid in acute caffeine intoxication: a case report.
Caffeine is arguably the most widely used stimulant drug in the world. Here we describe a suicide attempt involving caffeine overdose whereby the patient's severe intoxication was successfully treated with the prompt infusion of Intralipid. A 19-year-old man was found in an agitated state at home by the volunteer emergency team about 1 h after the intentional ingestion of 40 g of caffeine (tablets). His consciousness decreased rapidly, followed quickly by seizures, and electrocardiographic monitoring showed ventricular fibrillation. Advanced life support maneuvers were started immediately, with the patient defibrillated 10 times and administered 5 mg epinephrine in total and 300 + 150 mg of amiodarone (as well as lidocaine and magnesium sulfate). The cardiac rhythm eventually evolved to asystole, necessitating the intravenous injection of epinephrine to achieve the return of spontaneous circulation. However, critical hemodynamic instability persisted, with the patient's cardiac rhythm alternating between refractory irregular narrow complex tachycardia and wide complex tachycardia associated with hypotension. In an attempt to restore stability we administered three successive doses of Intralipid (120 + 250 + 100 mg), which successfully prevented a severe cardiovascular collapse due to a supra-lethal plasma caffeine level (>120 mg/L after lipid emulsion). The patient survived without any neurologic complications and was transferred to a psychiatric ward a few days later. The case emphasizes the efficacy of intravenous lipid emulsion in the resuscitation of patients from non-local anesthetic systemic toxicity. Intralipid appears to act initially as a vehicle that carries the stimulant drug away from heart and brain to less well-perfused organs (scavenging mechanism) and then, with a sufficient drop in the caffeine concentration, possibly as a tonic to the depressed heart.


Characterization of suppressive oligodeoxynucleotides that inhibit Toll-like receptor-9-mediated activation of innate immunity.
Synthetic oligodeoxynucleotides containing unmethylated CpG sequences (CpG-ODNs) stimulate Toll-like receptor-9 (TLR-9), thereby activating innate immunity. Stimulatory CpG-ODNs have been shown to be valuable in modifying immune responses in allergy, infection and cancer. Recently, it has been reported that the stimulation of TLR-9 by endogenous DNA might contribute to the pathogenesis of autoimmune diseases. We here report the identification of a suppressive, guanosine-rich ODN (G-ODN) that inhibited the activation of TLR-9 by stimulatory CpG-ODNs. The G-ODN was suppressive in murine macrophages and dendritic cells as well as in human plasmacytoid dendritic cells in vitro. G-ODN blocked the secretion of tumour necrosis factor-alpha (TNF-alpha) and interleukin-12p40 and interfered with the up-regulation of major histocompatibility complex (MHC) class II and costimulatory molecules. G-ODN was inhibitory even at a molar ratio of 1:10 (G-ODN:CpG-ODN) and when administered up to 7 hr after stimulation with CpG. G-ODN specifically inhibited TLR-9 but not other TLRs. Inhibition was dependent on a string of five guanosines. G-ODN was also inhibitory in an in vivo model of CpG/galactosamin (GalN) lethal shock. G-ODN interfered with upstream TLR-9 signalling. However, by extensive analysis we can exclude that G-ODN acts at the stage of cellular uptake. G-ODN therefore represents a class of suppressive ODNs that could be of therapeutic use in situations with pathologic TLR-9 activation, as has been proposed for certain autoimmune diseases.


Histomorphological findings and expression of matrix metalloproteinases and their tissue specific inhibitors (TIMPs) in normal tricuspid valves and in chronic tricuspid valvular disease in dogs.
The histomorphological findings and immunohistochemical expression of matrix metalloproteinases (MMPs-1, 2, 9 and 14) and their tissue inhibitors (TIMPs-2, 3 and 4) are reported in the parietal (pTV) and septal leaflets (sTV) of the tricuspid valves in normal dogs and dogs with chronic valvular disease (CVD). The layers of the normal sTV were not as well defined as in the pTV and the spongiosa of the sTV contained abundant mucopolysaccharides (MPS) and adipocytes. In CVD, there was expansion of the spongiosa of the pTV due to deposition of MPS, leading to formation of nodules along the free edge. In CVD, there was fibrosis of the atrialis of the sTV and formation of nodular deposits of MPS in the spongiosa and ventricularis, mainly affecting the proximal and middle parts of the leaflet. In dogs with normal pTV and sTV, MMPs-1 and 14 and TIMPs-2, 3 and 4 were expressed, while MMPs-2 and 9 were absent. In mild CVD, expression of MMPs-2, 9 and 14 were increased in the pTV, whereas small foci within the spongiosa contained MMP-9 and TIMP-3 positive cells. In advanced CVD, MMP-14 also was increased in the pTV. In mild CVD, there was increased expression of MMPs-1 and 2 and TIMP-2, but decreased expression of TIMP-4, in the sTV. Small foci with expression of MMP-14 and TIMPs-2, 3 and 4 were also present in the sTV in mild CVD. In advanced CVD, there was increased expression of MMPs-2 and 9 and TIMP-2 in the sTV. In CVD there is upregulation of various MMPs in the pTV, whereas there is a complex alteration in expression of MMPs and TIMPs in the sTV.


Assessment of the effects of non-phthalate plasticizer DEHT on the bivalve molluscs Mytilus galloprovincialis.
Due to their uncontrolled use, plastics has become an environmental concern, not only for their varying dimension but also for the potential release of substances such as phthalates (PAEs) and non-phthalates (NPPs) into the water. Phthalates are the most common plasticizers of concern, but non-phthalate plasticizers such as di (2-ethylhexyl) terephthalate (DEHT) have also been lately found in the marine environment. Mytilus galloprovincialis is a well-known bioindicator of aquatic environments due to its ability to accumulate a wide variety of xenobiotics, including plasticizers. Hence, aim of this study was to evaluate the potential bioaccumulation and effects of the NPP DEHT on M. galloprovincialis. To this purpose, following exposure to DEHT at 1 mg/l (DEHT1) and 100 mg/l (DEHT100), its accumulation in tissues and its effects on total lipids and fatty acid (FA) composition, protein content, cell viability, ability to recover volume and changes in biomarkers of oxidative stress were assessed. Mussels were able to bioaccumulate DEHT in their tissues, with a statistically significant increase compared to the control organisms. Differences in FA composition were observed after exposure, since C16:0, C18:0, C20:5ω-3 and C22:6ω-3 were significantly decreased from control to exposed groups. As a result, total SFA, MUFA and PUFA were affected in DEHT-exposed groups. Also, total protein varied following DEHT exposure, and significantly decreased in the DEHT100-group. Considering the physiological responses, both DEHT-exposed groups lost their ability to return to the original volume of digestive gland (DG) cells. On the other hand, oxidative biomarkers in the gills and DG were not significantly affected by the DEHT exposure. Overall, this study showed for the first time that DEHT exposure differentially affect mussels, in their lipid and protein metabolism, as well as cellular parameters.


Separation and characterization of underivatized oligosaccharides using liquid chromatography and liquid chromatography-electrospray ionization mass spectrometry.
Native cyclodextrin-based columns are particularly useful for the analysis of oligosaccharides because the retention of these carbohydrates is based mainly on the hydrogen bonding interactions of oligosaccharide hydroxyl groups with the stationary phase. Thus, the retention time predictably increases with the number of analyte hydroxyl groups, which corresponds to the elongation of the oligosaccharide chain. High-performance liquid chromatography (HPLC) coupled to electrospray ionization (ESI) mass spectrometry (MS) was used for the separation and characterization of underivatized oligosaccharide mixtures. With the limits of detection as low as 50 pg, all individual components of oligosaccharide mixtures (up to 11 glucose units long) were baseline resolved on a Cyclobond I 2000 column and detected using ESI-MS. Low flow rates and narrow I.D. columns increase the ESI-MS sensitivity significantly. The method showed potential usefulness for the sensitive and quick analysis of hydrolysis products of polysaccharides, and for trace levels of individual oligosaccharide or oligosaccharide isomers from biological systems.


Differences in spectral response properties of LGN cells in male and female squirrel monkeys.
The spectral response patterns of single cells recorded from the lateral geniculate nucleus (LGN) of the squirrel monkey (Saimiri sciureus) were examined in light of recent behavioral evidence indicating that there are significant variations in visual sensitivity and color vision within this species. The principal results from an analysis of data obtained from 1084 LGN cells were that (a) a significantly higher proportion of cells showing spectrally-opponent response patterns were recorded from female subjects, and (b) the proportion of spectrally-opponent units showing RG response patterns is very much higher in females than in males. These gender-related differences in the response patterns of LGN cells are discussed in the context of the behavioral results.


Influence of exogenous ATP on blood sugar, serum insulin and serum free fatty acids in short-term experimental hyperthyroid dogs and in euthyroid controls.
We studied the influence of ATP administration on blood sugar (BS), serum immunoreactive insulin (IRI) and serum free fatty acid (FFA) responses to glucose-induced hyperglycemia in short-term experimental hyperthyroid (STEH) dogs and in euthyroid controls. Hyperthyroidism was induced by a 10-day s.c. treatment with 100 micrograms/kg body weight 1-thyroxine. Glucose challenge was performed by i.v. infusion (700 mg/kg body weight as a priming dose, followed by 20 mg/kg body weight/min for 60 min). ATP (1 mg/kg body weight/min) was infused for 100 min 40 min before and 60 min during glucose administration). Glucose-induced hyperglycemia was more prolonged in the hyperthyroid group. ATP treatment did not affect the BS profile of controls but raised that of STEH animals. In the normal controls the insulinemic response to hyperglycemia was enhanced during ATP infusion. By contrast, in the STEH dogs insulin levels during glucose infusion was lower than in controls and did not significantly increase when ATP was added. ATP infusion induced a significant elevation of serum FFA, which was more pronounced in hyperthyroid animals with a greater fall during glucose administration and a marked increase during the period of recovery from hyperglycemia. In conclusion, we postulate that short-term hyperthyroidism in dogs may inhibit adenylate cyclase function in pancreatic B-cells and chiefly stimulate the action of cAMP-phosphodiesterase activity, thereby affecting insulin secretion.


Unenhanced Velocity-Selective MR Angiography (VS-MRA): Initial Clinical Evaluation in Patients With Peripheral Artery Disease.
Safe and accurate imaging of the peripheral arterial system is important for diagnosis and treatment planning of patients with peripheral artery disease (PAD). To evaluate image quality and diagnostic performance of unenhanced magnetic resonance angiography (MRA) based on velocity-selective (VS) magnetization preparation (termed VS-MRA). Prospective. Thirty-one symptomatic PAD patients underwent VS-MRA. Twenty-four of them underwent clinical digital subtraction angiography (DSA) examination, 18.8 ± 5.2 days after the MR scans. 1.5T MRI that included VS-MRA (homemade research sequence) and phase-contrast flow imaging (clinical sequence). Image quality (0: nondiagnostic, 3: excellent) and stenosis severity (0: normal, 3: occlusion) of VS-MRA images were assessed independently by three reviewers. Arterial signal-to-noise-ratio (SNR) and artery-to-muscle contrast-to-noise ratio (CNR) were calculated. The sensitivity and specificity of VS-MRA were calculated for the detection of significant stenosis (>50%) with DSA as the reference standard. Interobserver agreement among the three reviewers was evaluated by using Cohen κ-statistics. The image quality score of VS-MRA was 2.7 ± 0.5 for Reader 1, 2.8 ± 0.5 for Reader 2, and 2.8 ± 0.4 for Reader 3; SNR and CNR were 37.8 ± 12.5 and 30.5 ± 11.8, respectively. Segment-based analysis revealed that VS-MRA had sensitivities of 85.3%, 74.5%, and 78.4%, respectively, for the three reviewers, and specificities of 93.5%, 96.8%, and 95.2%. The interobserver agreement for the stenosis grading was good, as demonstrated by Cohen κ values of 0.76 (Reader 1 vs. Reader 2), 0.82 (Reader 1 vs. Reader 3), and 0.79 (Reader 2 vs. Reader 3). Unenhanced VS-MRA allows clear depiction of the peripheral arteries and accurate stenosis grading, as evidenced by high image quality scores and strong agreement with DSA. 2 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2019;49:744-751.


Structure and expression of a light-inducible shoot-specific rice gene.
By differential screening of a cDNA library of two-week-old rice seedlings cDNA clones were obtained, corresponding to shoot-specific mRNAs. By sequence analysis two of these clones were found to be rbcS cDNA clones. The mRNA corresponding to a third cDNA clone (COS5) displayed an expression pattern similar to the expression pattern of rbcS genes. The mRNA (800 bases) was light-inducible and encoded by a single-copy gene. The genomic clone (GOS5) was isolated and the intron/exon structure was determined by comparing the nucleotide sequences of the mRNA and the genomic clone. The gene contains two introns. Transcription start sites were determined by S1-nuclease mapping and primer extension. The start site obtained by both methods is located 87 bp upstream of the translation start site and 23 bp downstream of TATA box-like sequence. In the 5' non-coding region motifs can be found that are homologous to sequences in promoters that are light- or UV-inducible or confer leaf-specific expression. The open reading frame present in GOS5 codes for a protein (15 kDa) that contains a putative chloroplast transit peptide and does not show any significant homology to protein sequences in the NBRF protein database.


Form and accuracy of voluntary ocular tracking movements in response to sinusoidally moving acoustic targets.
Form and accuracy of voluntary ocular tracking in response to a horizontal sinusoidally moving auditory stimulus were examined in complete darkness. Forty young, healthy subjects participated in the study. Ocular movements under the applied experimental conditions consisted of a succession of saccades whose direction corresponded to the motion of sound. Though slight intersaccadic drifts were observed, pure smooth pursuit movements could not be recorded. A comparison of the amplitude of sound motion and the corresponding ocular motion showed the latter to be rather inaccurate. Independent of the actual amplitude of the sound motion (+/- 15 degrees up to +/- 90 degrees) the amplitude of the ocular movements, in most cases, remained constant within a range of approximately +/- 35 degrees to +/- 45 degrees. The accuracy in respect to phase relationship between target movement and ocular response was quite good. In response to sinusoidal motion of the sound, the ocular movements usually occurred simultaneously up to 0.5 Hz.


Effect of ultrasound-assisted osmotic dehydration on the drying kinetics, water state, and physicochemical properties of microwave vacuum-dried potato slices.
The effects of pre-treatments on the drying characteristics, water state, thermal properties, and bulk shrinkage of potato slices during microwave vacuum drying (MVD) were investigated. The pre-treatment included ultrasound in distilled water (USOD-0%), and ultrasound-assisted osmotic dehydration in a 60% sucrose solution (USOD-60%). Results showed that the drying time of potato slices was reduced and the drying rate was increased when USOD-0% was used as a pretreatment, whereas USOD-60% had a negative effect on the drying rate of the samples. The Weibull model was effective in predicting the water changes in potato slices during the drying process. NMR analysis revealed that the relative content of immovable water (M22) increased initially, then decreased for drying, while the transverse relaxation time (T2) and the relative content of free water (M23) decreased consistently. The DSC results indicated that the glass transition temperature (Tg) had an inverse relationship with the water content of the samples, yet had virtually no influence on the volume shrinkage. The sample volumes decreased linearly with the decrease in water content during the initial drying stages. USOD pre-treatment lessened the volume shrinkage of MVD potato slices. Static gravimetry was used to determine the moisture sorption isotherms of MVD potato slices at 30 °C within the aw range of 0.113-0.923. The GAB model accurately fitted the experimental sorption data, which showed sigmoid shape curves for the MVD samples. When aw values exceeded 0.7, the USOD-60% treatment significantly reduce the water sorption capacity of MVD potato slices, while USOD-0% treatment was observed to increase the hygroscopic properties of MVD samples. Compared with the control, USOD-0% pretreatment significantly increased the monolayer water content (X0), sorption surface area (S0), the thickness of sorbed water multilayer (tm), and density of sorbed water (Ds) values of MVD potato slices, while USOD-60% decreased these values.


Suicide in India during the first year of the COVID-19 pandemic.
There has been substantial discussion as to whether the mental health and socio-economic consequences of the COVID-19 pandemic might impact suicide rates. Although India accounts for the largest proportion of global suicides, the early impacts of the COVID-19 pandemic on suicide rates in this country are unknown. National Crime Records Bureau (NCRB) data were used to calculate annual suicide rates for the period 2010-2020, stratified by sex and state. Rate Ratios (RRs) stratified by sex and state were calculated to estimate the extent of change in suicide rates. Suicide rates in India generally showed a decreasing trend from 2010 until 2017, with the trend reversing after this period, particularly for males. Among males and females, the highest increase post 2017 was noted in 2020 (compared to 2017) (males: RR = 1.18 95% UI 1.17-1.19; females: RR = 1.05 95% UI 1.03-1.06). Suicide rates based on the NCRB data might be an underestimation of the true suicide rates. Suicide rates in India increased during the first year of the COVID-19 pandemic, and although the increase in suicide rates, especially among males, predates the pandemic, the increase in suicide rates was highest in 2020, compared to increases in previous years. Further research is warranted to understand the potential ongoing impact of the COVID-19 pandemic on suicide in India.


Attentional bias for affective visual stimuli in posttraumatic stress disorder and the role of depression.
An attentional bias for trauma-related verbal cues was frequently demonstrated in posttraumatic stress disorder (PTSD) using variants of the emotional Stroop task (EST). However, the mechanisms underlying the Stroop-effect are ill-defined and it is yet unclear how the findings apply to different paradigms and stimulus modalities. To address these open questions, for the first time a spatial-cuing task with pictorial cues of different emotional valence was administered to trauma-exposed individuals with and without PTSD, and non-trauma-exposed controls. Groups did not show different response profiles across affective conditions. However, a group effect was evident when comparing depressed with non-depressed individuals: Those with depression showed delayed attending towards trauma-related cues and faster attending away from negative cues. In correlational analyses, attentional avoidance was associated with both depression and PTSD symptom severity. These findings highlight the need for research on trauma populations and anxiety in general to pay closer attention to depression as an important confound in the study of emotional information processing.


DNA topology: topological transformation of DNA by reaction of cisplatin-DNA-core histone complexes with human DNA topoisomerase.
Cis-diamminedichloroplatinium(II), an active antitumor agent binds to core-histone -SV40 DNA complexes prepared by reaction of DNA with core-histone, and alters the fiber-like structure into the loosened structure. In order to model the cisplatin-modified chromatin complexes in cell, the complexes were reacted with human DNA topo II. We found the generations of unique topologically isomers such as trefoil knot (and catenane) and pseudo catenane (and pseudo knot) by reaction of cis-DDP--linearDNA--core-histone complexes with DNA topo I. The results are discussed in relation with a possible recombinational role of topo II (or topo I) on the reaction with cis-DDP--DNA--core-histone complexes.


Identification of two distinct chromosome 12-derived amplification units in neuroblastoma cell line NGP.
The neuroblastoma cell line NGP contains two homogeneously staining regions (hsr). One of these hsrs contains MYCN sequences. Reverse painting experiments demonstrated that the second HSR consisted of two chromosome 12-derived amplification units, located at 12q14-15 and 12q24. Southern blot and fluorescence in situ hybridization (FISH) analysis showed amplification of genes located at 12q14-15: SAS, MDM2, and CDK4, GLI, CHOP, CDK2, and A2MR were found not to be amplified. FISH further demonstrated amplification of RSN, a gene located at 12q24. The finding of two distinct chromosome 12 amplification units in a neuroblastoma cell line NGP is reminiscent of recent findings in well-differentiated liposarcoma (WDLPS) and other sarcomas. The second amplification unit on chromosome 12 in NGP is located more distal (12q24) than the one observed in WDLPS (12q21). The mechanism and biologic significance of this amplification process in neuroblastoma and WDLPS remain to be elucidated.


A model for anomalous rectification: electrochemical-potential-dependent gating of membrane channels.
A model is presented for "anomalous rectification" based upon electrical measurements on the egg cell membrane of the starfish. The objective is to postulate a plausible molecular mechanism which yields an expression for the conductance similar to that deduced empirically by Hagiwara and Takahashi (1974), i.e.,: formula: (see text), where B, deltaVh and v are constant, cK is the external K+ concentration, and deltaV (= V - V0) is the displacement of the membrane potential from its resting value. It is shown that a similar dependence of the conductance on deltaV is expected for a particular class of models in which the K+ ions are also implicated in "gating". To give a specific example, we consider the case in which the formation of ion-permeable pores requires a voltage-induced orientation of membrane-bound, electrically-charged groups and subsequent complexation of these groups with the external cations. Furthermore, the proportionality between GK and CK1/2, when the internal K+ concentration is constant, is accounted for by conventional descriptions of the ionic fluxes using Eyring's rate reaction theory. In terms of the present model, B and deltaVh are explicit functions of the internal K+ concentrations and are thus constant only as long as this is unvaried. The particular value of v required to fit the data (v congruent to 8.4 mV) is rationalized by the assumption that each of the orientable groups carries three negative elementary charges. In addition, the predictions of the present model are compared with those deduced from an alternative viewpoint, which is related to Armstrong's "blocking particle hypothesis", in that the probability for opening and closing of the pore is assumed to depend on whether the pore is occupied or empty. Differences and similarities between the two models, as well as ways to discriminate between them, are discussed.


High resolution radioautographic investigation of [3H]GABA accumulating neurons in cat sensorimotor cortical areas.
After the injection of [3H]GABA in the cat sensorimotor cortex, radioautographic analysis of thick and thin sections showed an intense labeling of numerous nerve fibers and of some neuronal cell bodies. Labeled boutons displayed either only clear vesicles or clear and large granular vesicles. They could constitute symmetrical synaptic differentiations upon dendrites or dendritic spines. Radioactive perikarya of stellate neurons could be labeled either by direct uptake or by retrograde axonal flow.


Resuscitation factors from mycobacteria: homologs of Micrococcus luteus proteins.
Resuscitation promoting factors (Rpfs) are proteins, originally identified in Micrococcus luteus, that promote recovery of bacteria from a viable but non-replicating phase (e.g., stationary phase or latency) to a replicating phase. Purified M. luteus Rpf can stimulate growth and increase recovery of M. luteus bacteria as well as Mycobacterium tuberculosis bacteria from prolonged stationary cultures. To clone and characterize Rpfs from mycobacteria. We cloned one M. avium subsp. paratuberculosis rpf gene and one M. tuberculosis rpf gene into the pET19b or pET21a vector for expression in Escherichia coli. The His-tag recombinant proteins were purified and characterized. When the purified recombinant proteins were added to Sauton medium (a relatively minimal medium) at 100-500 pM, lag phase for mycobacteria from non-replicating cultures was shortened and there was a 10- to 100-fold increase in colony-forming units compared with control samples. In most probable number assays, the mycobacterial Rpfs increased recovery of mycobacteria from late stationary culture by about 10-fold. The Rpfs also promoted recovery of extensively washed Mycobacterium smegmatis bacteria inoculated into Sauton medium. Rpfs had only minor effects on growth of M. tuberculosis in BACTEC 12B broth, a rich medium. The mycobacterial Rpfs demonstrate resuscitation activities similar to those of the M. luteus Rpf.


High-dose-rate brachytherapy for prostate cancer in a previously radiated patient with polyethylene glycol hydrogel spacing to reduce rectal dose: case report and review of the literature.
To describe the use of a temporary spacer to reduce rectal dose prior to prostate radiation in a man with prior pelvic radiotherapy and review the relevant literature. A healthy 57-year-old man presented with high-risk prostate cancer (Gleason score of 8, prostate-specific antigen level [PSA] 12.6 ng/mL, T3a by magnetic resonance imaging [MRI]), only 2.5 years after a low anterior resection followed by pelvic chemo-radiotherapy to 50.4 Gy for a locally advanced rectal cancer. Due to the prior radiation, he was not felt to be a candidate for surgery or external beam radiation, so he chose long-term androgen deprivation therapy (ADT) plus high-dose-rate brachytherapy to 36 Gy in 6 fractions. To reduce the radiation dose to the anterior rectal wall, 10 mL of a polyethylene glycol hydrogel spacer was injected between the prostate and rectum and created between 1.4 and 1.5 cm of separation along the length of the prostate. Two randomized trials demonstrating that local therapy plus ADT improves overall survival compared to ADT alone provided the rationale for additional prostate radiotherapy in this otherwise healthy patient. Salvage brachytherapy is associated with a 3.4% rate of rectal fistula among the 251 cases reported in the literature from 2000-2007, with rates as high as 12% in one series. The spacer allowed the rectal dose constraint goals to be easily met. Injecting an absorbable polyethylene glycol hydrogel to separate the prostate and rectum appears to be associated with decreased maximum and mean rectal doses, and may have particular utility in previously irradiated patients.


Nanoanesthesia: a novel, intravenous approach to ankle block in the rat by magnet-directed concentration of ropivacaine-associated nanoparticles.
As an alternative to current methods of local nerve block, we studied the feasibility of producing ankle block in the rat with IV injection of magnetic nanoparticles (MNPs) associated with ropivacaine and application of a magnet at the ankle. The anesthetic effect of magnet-directed ropivacaine-associated MNPs (MNP/Ropiv) was tested in the rat using paw withdrawal latencies from thermal stimuli applied to the hindpaw. The MNP/Ropiv complexes consisted of 0.7% w/v ropivacaine and 0.8% w/v MNPs containing 12% w/w magnetite (F₃O₄). The effect of IV injection of MNP/Ropiv with 15, 30, and 60-minute magnet application to the right ankle was compared with the effect without magnet application on the left hindpaw, to conventional ankle block with 0.1% or 0.2% ropivacaine, and to IV injection of MNPs alone with 30-minute magnet application to the right ankle. In addition, the pharmacokinetics of the MNP/Ropiv complexes were determined. IV injection of MNP/Ropiv with magnet application at the ankle significantly increased paw withdrawal latencies from thermal stimuli compared with pretreatment baselines in the same paw (P < 0.0001) and compared with the contralateral paw without magnet application (P < 0.0001). IV injection of MNPs alone had no significant effect on paw withdrawal latency. Absolute ropivacaine concentrations in ankle tissue, and ankle tissue-to-plasma concentration ratios were higher in the MNP/Ropiv group with 30-minute magnet application compared with MNP/Ropiv group without magnet application (mean ± SEM, 150 ± 10 ng/g vs 105 ± 15 ng/g, respectively, and 6.1 ± 0.8 vs 4.2 ± 0.7, respectively). The current study establishes proof of principle that it is possible to produce ankle block in the rat by IV injection of MNP/Ropiv complexes and magnet application at the ankle. The results indicate that further study of this approach is warranted.


Ambivalence of nonoffending guardians after child sexual abuse disclosure.
A concern in the intervention with sexually abused children is the support of their nonoffending guardians after disclosure of the abuse. Approximately a third of nonoffending guardians respond with vacillation in support, and these nonoffending guardians are at greater risk for having their children removed. This article reconceptualizes vacillation in support as an ambivalent response. Drawing on the interdisciplinary literature, this article suggests that ambivalence in support reflects the confluence between the nonoffending guardian's valence toward the child and perpetrator. This article further proposes that ambivalence is normative when the costs of disclosure are high and when the nonoffending guardian is ambivalent/preoccupied in attachment. Ambivalence may also be both a precursor to and an effect of the traumatic experience of the disclosure on the nonoffending guardian. In a study of 30 nonoffending mothers whose partners sexually abused their children, these relationships were supported.


Continuing inequality: gender and social class influences on self perceived health after a heart attack.
To investigate the effect of social class and gender on self perceived health status for those recovering from an acute myocardial infarction. A longitudinal survey design was used, collecting both qualitative and quantitative data. Quantitative data are reported in this article, obtained by questionnaire over the first year after the event. SF-36 and EQ-5D (EuroQol) were used to measure self perceived health status. Community based study in a city in the north of England. A consecutive sample of 229 people discharged from hospital after acute myocardial infarction. Overall gain in health status was found to be statistically significant over the year. Improvements were greatest in domains relating to role fulfillment and pursuit of normal and social activities. When analysed by gender, women showed poorer improvement than men, particularly in the domains relating to physical and social functioning. Analysed by social class, those without educational qualifications showed poorer improvement in pain experience and vitality. Access to a car was significant in avoiding physical limitations and promoting general health. Existing gradients between the health of women and men, and between the social classes, are maintained and probably exacerbated by the experience of acute illness, and health professionals need to be made aware of social groups who are at risk of poor rehabilitation.


Urinary incontinence between 12 and 24 months postpartum: a cross-sectional study nested in a Brazilian cohort from two cities with different socioeconomic characteristics.
We aimed to identify the prevalence of urinary incontinence (UI) during the postpartum period (12-24 months) in two cities with different socioeconomic indicators in Brazil (Ribeirão Preto, SP, and São Luís, MA) and to determine associated risk factors. A cross-sectional study nested in the Brazilian Ribeirão Preto and São Luís Birth Cohort Studies (BRISA) cohort was conducted in two Brazilian municipalities (Ribeirão Preto, and São Luís). A total of 13,214 women delivered in both cities (2010-2011). We interviewed 3,751 postpartum women in Ribeirão Preto and 3275 in São Luís (2011-2013). Univariate and multivariate analyses were performed to assess factors associated with postpartum UI for each municipality. The prevalence of self-reported UI at 12-24 months postpartum was 16.3% (611 out of 3,751) in Ribeirão Preto and 11.4% (375 out of 3,275) in São Luís (p < 0.001). The univariate analysis performed at Ribeirão Preto showed that women who were obese, who had diabetes or gestational diabetes and who presented with excessive weight gain during gestation presented an association with postpartum UI. However, only weight gain during pregnancy remained strongly associated with UI on multivariate analysis (p = 0.009; OR 1.041 [1.010-1.073]). On the other hand, in São Luís, no risk factors were associated with postpartum UI at univariate and multivariate analysis. The prevalence of UI was higher in Ribeirão Preto (higher socioeconomic level) than in São Luís. Weight gain during pregnancy was statistically associated with UI in Ribeirão Preto. No independent variables remained associated in the final model with UI in São Luís.


Effects of dietary supplementation of probiotic and Spirulina platensis microalgae powder on growth performance immune response, carcass characteristics, gastrointestinal microflora and meat quality in broilers chick.
With the potential development of human pathogenic bacteria resistant to antibiotics, the use of antibiotics as growth promoter in poultry production was banned in different countries, and it has forced the poultry industry to consider 'Biologically safer' alternatives to antibiotics, among which the probiotics and microalgae can be mentioned. Present study aimed to compare Spirulina platensis microalgae in combination with a native probiotic as an alternative to antibiotics. 336 male broiler chicks were allotted into 7 treatments and 4 repetitions in a completely randomised design to evaluate chick's performance and immune response to different treatment based on indexes as feed intake, weight gain, feed conversion ratio, humoral immunity, carcass characteristics, thigh and breast pH, intestinal morphology and microbial population. European production efficiency coefficient was also reported. No significant difference was appeared in the pH of thigh and breast meat (p > 0.05). Supplementation of diets with SP0.3 revealed better villi height, villi length to crypt depth ratio and villi surface. With significant difference (p < 0.05), the highest and lowest colonies of Lactobacillus and E. coli were recorded for PR0.5 SP0.3 treatments. Supplementation of broilers diets either with probiotic prepared from the microorganism isolated of native birds (1 g/kg) or S. platensis (0.2 g/kg) alone and their combination (0.3 g/kg of S. platensis in combination with 0.5 g/kg of native probiotic) are promising and can be a good alternative to antibiotics, lead to progress of broiler's performance.


Neuronal contact upregulates astrocytic sphingosine-1-phosphate receptor 1 to coordinate astrocyte-neuron cross communication.
Astrocytes, the most abundant glial cells in the mammalian brain, directly associate with and regulate neuronal processes and synapses and are important regulators of brain development. Yet little is known of the molecular mechanisms that control the establishment of astrocyte morphology and the bi-directional communication between astrocytes and neurons. Here we show that neuronal contact stimulates expression of S1PR1, the receptor for the bioactive sphingolipid metabolite sphingosine-1-phosphate (S1P), on perisynaptic astrocyte processes and that S1PR1 drives astrocyte morphological complexity and morphogenesis. Moreover, the S1P/S1PR1 axis increases neuronal contact-induced expression of astrocyte secreted synaptogenic factors SPARCL1 and thrombospondin 4 that are involved in neural circuit assembly. Our findings have uncovered new functions for astrocytic S1PR1 signaling in regulation of bi-directional astrocyte-neuron crosstalk at the nexus of astrocyte morphogenesis and synaptogenesis.