Oral Midazolam Vs Promethazine as Pre Sedation Medication in Pediatric Dentistry

Objectives Pre- and post-sedation effect of oral Midazolam to promethazine in2-6 yrs old fearful children for dental treatmentMethods This randomized clinical trial was carried out on a group of 26 children aged 2-6 years referred to the dental school due to their fear and multiple dental needs. Patients were selected from ASA I or II classification and scored 1 in Frankl Behavior scale. Each patient was scheduled for two subsequent visits to receive one of the two pre medications before IV sedation. Each patient served as self-control and randomly assigned to either group A: receiving Midazolam oral as premed in 1st visit or group B: receiving Promethazine oral as the premed in 1st visit. Six hour NPO was instructed prior to sedation visit. Monitoring vital signs were conducted at every 15 minutes starting with base line before any drug administration. Sedation score was recorded using Houpt Sedation scale. Post sedation problems were recorded by operator. Data were analyzed using Student t test and Kruskal Wallis.Results No significant difference was noted between the patient perceptions at the two different visits. Children did not show a significant difference on symptoms such as Crying, Movement, Sleep and overall behavior in two visits at the first 15 minutes of sedative injection. Post-operative complications were having no significant difference. Lower sickness and vomiting were reported following promethazine intake.Conclusion Promethazine seems to be as effective and as acceptable premedication as Midazolam in pediatric dentistry

Designing novel construction for cell surface display of protein E on Escherichia coli using non-classical pathway based on Lpp-OmpA

Additional file 2. pNon-OmpA-PE. ab1. Sequencing result of pNon-OmpA-PE

Exploring the interplay between Fusobacterium nucleatum with the expression of microRNA, and inflammatory mediators in colorectal cancer

BackgroundFusobacterium nucleatum has been recognized as an important key bacterium in the cause and spread of colorectal carcinogenesis. Nevertheless, the clinical relevance of F. nucleatum in colorectal cancer (CRC) and its effect on immune factors and the tumor microenvironment have not been fully elucidated.Materials and methodsThe frequency of F. nucleatum was measured in 100 paired tumor and normal tissue specimens by TaqMan quantification Real-Time Polymerase Chain Reaction (qPCR). The mRNA expression levels of cytokines (IL-6, IL-10, IL-12β, IL-17, TNF-α, TLR-2, and TLR-4), and miRNAs (miR-21, miR-31) were examined. Eventually, any potential correlations between the molecular and clinicopathological features of the neoplastic samples and the abundance of F. nucleatum were analyzed.ResultsThe relative frequency of F. nucleatum was significantly increased in cancerous tissue compared to adjacent non-tumor tissues. Furthermore, the high level of F. nucleatum was significantly associated with histological grade III and IV CRC tissues (P = 0.027 and P = 0.022, respectively) and perineural invasion-positive patients (P = 0.037). In addition, the expression levels of IL-6, IL-17, TNF-α,IL-12β, TLR-2, and TLR-4 as well as miR-21 and miR-31 showed a significant increase in the cancer group. A notable correlation was also observed between the high status of F. nucleatum and the expression of IL-6, TNF-α and miR-21.ConclusionOur results emphasize the importance of F. nucleatum and changes in the expression of genes involved in CRC. Studying the microbial profile and gene expression changes in CRC patients may be a promising approach to improve screening methods and provide therapeutic strategies

A Classic Case of Maple Syrup Urine Disease and a Novel Mutation in the BCKDHA Gene

Background: Maple syrup urine disease (MSUD) is an inherited branched-chain amino acid metabolic disorder caused by the deficiency in the branched-chain alpha-keto acid dehydrogenase (BCKD) complex. In MSUD, elevation of the branched-chain amino acids, such as alpha-keto acid and alpha-hydroxy acid, occurs due to the BCKDC gene deficiency, appearing in the blood, urine, and cerebrospinal fluid, which leads to neurological damage and mental retardation. MSUD phenotypically penetrates due to the mutations in the coding genes of four subunits of the BCKD complex, including the BCKDHA, BCKDHB, DBT, and DLD genes.Case report: We aimed to report the cases of three families whose children were affected by MSUD and presented with symptomatic features during the first week of birth, which were identified by mass spectrometry. DNA study was performed as a diagnosis panel containing four encoded BCKDC subunit genes.Conclusion: In the current study, DNA analysis and phenotypic manifestations indicated a novel mutation of c.143delT, p.L48Rfs*15 in the BCKDHA gene in a homozygous state, which is a causative mutation for the classic MSUD phenotype. Early diagnosis and neonatal screening are recommended for the accurate and effective treatment of this diseas

ActuAtor, a Listeria-inspired molecular tool for physical manipulation of intracellular organizations through de novo actin polymerization

細胞の中のものを「押す」方法を開発 --細胞内構造体の“かたち”と機能の関係を明らかに--. 京都大学プレスリリース. 2023-09-25.Form and function are often interdependent throughout biology. Inside cells, mitochondria have particularly attracted attention since both their morphology and functionality are altered under pathophysiological conditions. However, directly assessing their causal relationship has been beyond reach due to the limitations of manipulating mitochondrial morphology in a physiologically relevant manner. By engineering a bacterial actin regulator, ActA, we developed tools termed “ActuAtor” that inducibly trigger actin polymerization at arbitrary subcellular locations. The ActuAtor-mediated actin polymerization drives striking deformation and/or movement of target organelles, including mitochondria, Golgi apparatus, and nucleus. Notably, ActuAtor operation also disperses non-membrane-bound entities such as stress granules. We then implemented ActuAtor in functional assays, uncovering the physically fragmented mitochondria being slightly more susceptible to degradation, while none of the organelle functions tested are morphology dependent. The modular and genetically encoded features of ActuAtor should enable its application in studies of the form-function interplay in various intracellular contexts

From bedside to bench and back again: translational studies of mechanical unloading of the left ventricle to promote recovery after acute myocardial infarction [version 1; referees: 2 approved]

Heart failure is a major cause of global morbidity and mortality. Acute myocardial infarction (AMI) is a primary cause of heart failure due in large part to residual myocardial damage despite timely reperfusion therapy. Since the 1970’s, multiple preclinical laboratories have tested whether reducing myocardial oxygen demand with a mechanical support pump can reduce infarct size in AMI. In the past decade, this hypothesis has been studied using contemporary circulatory support pumps. We will review the most recent series of preclinical studies in the field which led to the recently completed Door to Unload ST-segment Elevation Myocardial Infarction (DTU-STEMI) safety and feasibility pilot trial

The Heart Is an Early Target of Anthrax Lethal Toxin in Mice: A Protective Role for Neuronal Nitric Oxide Synthase (nNOS)

Anthrax lethal toxin (LT) induces vascular insufficiency in experimental animals through unknown mechanisms. In this study, we show that neuronal nitric oxide synthase (nNOS) deficiency in mice causes strikingly increased sensitivity to LT, while deficiencies in the two other NOS enzymes (iNOS and eNOS) have no effect on LT-mediated mortality. The increased sensitivity of nNOS−/− mice was independent of macrophage sensitivity to toxin, or cytokine responses, and could be replicated in nNOS-sufficient wild-type (WT) mice through pharmacological inhibition of the enzyme with 7-nitroindazole. Histopathological analyses showed that LT induced architectural changes in heart morphology of nNOS−/− mice, with rapid appearance of novel inter-fiber spaces but no associated apoptosis of cardiomyocytes. LT-treated WT mice had no histopathology observed at the light microscopy level. Electron microscopic analyses of LT-treated mice, however, revealed striking pathological changes in the hearts of both nNOS−/− and WT mice, varying only in severity and timing. Endothelial/capillary necrosis and degeneration, inter-myocyte edema, myofilament and mitochondrial degeneration, and altered sarcoplasmic reticulum cisternae were observed in both LT-treated WT and nNOS−/− mice. Furthermore, multiple biomarkers of cardiac injury (myoglobin, cardiac troponin-I, and heart fatty acid binding protein) were elevated in LT-treated mice very rapidly (by 6 h after LT injection) and reached concentrations rarely reported in mice. Cardiac protective nitrite therapy and allopurinol therapy did not have beneficial effects in LT-treated mice. Surprisingly, the potent nitric oxide scavenger, carboxy-PTIO, showed some protective effect against LT. Echocardiography on LT-treated mice indicated an average reduction in ejection fraction following LT treatment in both nNOS−/− and WT mice, indicative of decreased contractile function in the heart. We report the heart as an early target of LT in mice and discuss a protective role for nNOS against LT-mediated cardiac damage

DE NOVO ENGINEERING OF CHEMOTACTIC TRANSDUCTION IN CELLS & CELL-MIMETIC DEVICES

Spatiotemporal regulation of signaling is an integral feature of biologicalnetworks. Cellular processes driven by membrane-localized forces are prevalentand crucial in both health and disease. These include cell division, chemo-taxis, and phagocytosis. For these processes to transpire, biochemical eventsleading to actin-polymerization at the plasma membrane are critical. Exactlyhow these reactions are triggered, the activation time-scale, and the molecu-lar players involved are not yet known. This is largely due to the complexityand redundancy in signaling networks, rendering our current biology tools in-adequate in extracting the key cell signaling features. Thus, this dissertationexplores a novel synthetic biology approach: we engineer two new cell-basedand cell-free platforms equipped with spatiotemporal control modules to probeactin polymerization events in the context of chemotaxis.One key molecule in the chemotactic pathway is the Rac1 GTPase that ac-tivates downstream signaling leading to actin polymerization and later motility.We coupled a Rac1 activator to a chemically inducible actuation unit to spa-tiotemporally regulate its activity. Using this paradigm, we engineered logiccomputation in living mammalian cells. We next triggered a Rac1-dependentsignaling in giant vesicles in order to achieve cell-like functionin vitro.ii We first devised Boolean negation computation in living mammalian cells.To enhance input specificity, we coupled the system to binary chemical inputsand engineered N-IMPLY and the universal, NAND and NOR logic. The dy-namic actin structures at the membrane served as the devices’ output. Thetime-scale of computation was few minutes, two orders of magnitude fasterthan the previously reported counterparts. Taken together, by deconstructionof cell signaling we engineered biocomputing and probed the spatiotemporalregulation of Rac1 activation and the ensuing membrane deformation.Next, we aimed to further reduce the Rac1 pathway to deduce the exactsignaling map. To this end, we studied Rac1 signaling and actin polymerizationin the context of chemotaxis through a reductionist lens: engineering an artificialcell that is built one molecule type at a time, is minimal in the number ofprotein and lipid components, and can be chemically induced to form actinstructures at the membrane. We adopted giant vesicles as “artificial cells” wherewe reconstituted thede novochemical sensing, and Rac1 signaling modules.This minimal proto-cell offer a new platform for studying signaling as ti bridgesthe gap between cell-based studies and thein vitroplatforms that lack thegeometric and dimension consideration. Also in the long term this class ofstudied could be used to unequivocally resolve the molecular players of actin-mediated processes. It could also serve as a biomimetic device that can bedeployed in a cellular environment to deliver drugs or genes.With the cell-based devices, expanding on the repertoire of signal transduc-tion computers requires formidable protein engineering efforts that will revealmuch about the signaling proteins’ structures and functions. With the cell-freedevices, beyond the biomimetics scope, this platform holds great potential foriii unraveling much about the role of lipids and proteins in signaling

DE NOVO ENGINEERING OF CHEMOTACTIC TRANSDUCTION IN CELLS & CELL-MIMETIC DEVICES

Spatiotemporal regulation of signaling is an integral feature of biologicalnetworks. Cellular processes driven by membrane-localized forces are prevalentand crucial in both health and disease. These include cell division, chemo-taxis, and phagocytosis. For these processes to transpire, biochemical eventsleading to actin-polymerization at the plasma membrane are critical. Exactlyhow these reactions are triggered, the activation time-scale, and the molecu-lar players involved are not yet known. This is largely due to the complexityand redundancy in signaling networks, rendering our current biology tools in-adequate in extracting the key cell signaling features. Thus, this dissertationexplores a novel synthetic biology approach: we engineer two new cell-basedand cell-free platforms equipped with spatiotemporal control modules to probeactin polymerization events in the context of chemotaxis.One key molecule in the chemotactic pathway is the Rac1 GTPase that ac-tivates downstream signaling leading to actin polymerization and later motility.We coupled a Rac1 activator to a chemically inducible actuation unit to spa-tiotemporally regulate its activity. Using this paradigm, we engineered logiccomputation in living mammalian cells. We next triggered a Rac1-dependentsignaling in giant vesicles in order to achieve cell-like functionin vitro.ii We first devised Boolean negation computation in living mammalian cells.To enhance input specificity, we coupled the system to binary chemical inputsand engineered N-IMPLY and the universal, NAND and NOR logic. The dy-namic actin structures at the membrane served as the devices’ output. Thetime-scale of computation was few minutes, two orders of magnitude fasterthan the previously reported counterparts. Taken together, by deconstructionof cell signaling we engineered biocomputing and probed the spatiotemporalregulation of Rac1 activation and the ensuing membrane deformation.Next, we aimed to further reduce the Rac1 pathway to deduce the exactsignaling map. To this end, we studied Rac1 signaling and actin polymerizationin the context of chemotaxis through a reductionist lens: engineering an artificialcell that is built one molecule type at a time, is minimal in the number ofprotein and lipid components, and can be chemically induced to form actinstructures at the membrane. We adopted giant vesicles as “artificial cells” wherewe reconstituted thede novochemical sensing, and Rac1 signaling modules.This minimal proto-cell offer a new platform for studying signaling as ti bridgesthe gap between cell-based studies and thein vitroplatforms that lack thegeometric and dimension consideration. Also in the long term this class ofstudied could be used to unequivocally resolve the molecular players of actin-mediated processes. It could also serve as a biomimetic device that can bedeployed in a cellular environment to deliver drugs or genes.With the cell-based devices, expanding on the repertoire of signal transduc-tion computers requires formidable protein engineering efforts that will revealmuch about the signaling proteins’ structures and functions. With the cell-freedevices, beyond the biomimetics scope, this platform holds great potential foriii unraveling much about the role of lipids and proteins in signaling

Effect of continuous aerobic training versus high intensity interval training on Resistin and insulin resistance in type 2 diabetic rats

Background: the adipose tissue produces and releases peptides that contribute to various processes in body, including insulin resistance. The aim of this study was to investigate the effect of eight weeks of continuous aerobic training and high intensity interval training on Resistin and insulin levels and insulin resistance in type 2 diabetic male wistar rats. Methods: Twenty-four Wistar rats became diabetic in seven months. In next phase, after introducing with training environment, Wistar rats were randomly assigned into three equal groups of six each: control, continuous (20 minutes, 60% maximum speed) and intense interval (2 minutes of activity with 80% maximum speed, 2 minutes recovery with 30% maximum speed). The rats trained five time a week for eight weeks. Resistin gene expression and plasma insulin and glucose levels were measured before and after eight weeks. One-way ANOVA was used at P<0.05 for statistical analysis of data. Results: regardless the type of training, differences between pre and post training results was statistically significant for insulin, glucose, insulin resistance and Resistin (P<0.05). Interval training lead to significant changes in all factors except the Resistin gene expression (P<0.05). There was a significant relationship between changes in insulin resistance and Resistin gene expression (r=0.63). Conclusion: The results of this study showed that training is an effective factor in insulin resistance process and related factors in diabetes, and Resistin also play a role in this process, but it seems that regular training is more important factor than its type to change the Expression of Resistin