Detection of antibacterial activity of essential oil components by TLC-bioautography using luminescent bacteria

The aim of the present study was the chemical characterization of some medically relevant essential oils (tea tree, clove, cinnamon bark, thyme and eucalyptus) and the investigation of antibacterial effect of the components of these oils by use of a direct bioautographic method. Thin layer chromatography (TLC) was combined with biological detection in this process. The chemical composition of the oils was determined by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Eucalyptol (84.2%) was the main component of the essential oil of eucalyptus, eugenol (83.7%) of clove oil, and trans-cinnamic aldehyde (73.2%), thymol (49.9%) and terpinen-4-ol (45.8%) of cinnamon bark, thyme and tea tree oils, respectively. Antibacterial activity of the separated components of these oils, as well as their pure main components (eucalyptol, eugenol, trans-cinnamic aldehyde and thymol) was observed against the Gram-negative luminescence tagged plant pathogenic bacterium Pseudomonas syringae pv. maculicola (Psmlux) and the Gram-negative, naturally luminescent marine bacterium Vibrio fischeri. On the whole, the antibacterial activity of the essential oils could be related to their main components, but the minor constituents may be involved in this process. Trans-cinnamic aldehyde and eugenol were the most active compounds in TLC-bioautography. The sensitivity of TLC-bioautographic method can be improved with using luminescent test bacteria. This method is more cost-effective and provides more reliable results in comparison with conventional microbiological methods, e.g. disc-diffusion technique

Uncoupling the excitatory amino acid transporter 2 from its C-terminal interactome restores synaptic glutamate clearance at corticostriatal synapses and alleviates mutant huntingtin-induced hypokinesia

Rapid removal of glutamate from the sites of glutamate release is an essential step in excitatory synaptic transmission. However, despite many years of research, the molecular mechanisms underlying the intracellular regulation of glutamate transport at tripartite synapses have not been fully uncovered. This limits the options for pharmacological treatment of glutamate-related motor disorders, including Huntington’s disease (HD). We therefore investigated the possible binding partners of transgenic EAAT2 and their alterations under the influence of mutant huntingtin (mHTT). Mass spectrometry analysis after pull-down of striatal YFP-EAAT2 from wild-type (WT) mice and heterozygote (HET) Q175 mHTT-knock-in mice identified a total of 148 significant (FDR < 0.05) binders to full-length EAAT2. Of them 58 proteins exhibited mHTT-related differences. Most important, in 26 of the 58 mHTT-sensitive cases, protein abundance changed back toward WT levels when the mice expressed a C-terminal-truncated instead of full-length variant of EAAT2. These findings motivated new attempts to clarify the role of astrocytic EAAT2 regulation in cortico-basal movement control. Striatal astrocytes of Q175 HET mice were targeted by a PHP.B vector encoding EAAT2 with different degree of C-terminal modification, i.e., EAAT2-S506X (truncation at S506), EAAT2-4KR (4 lysine to arginine substitutions) or EAAT2 (full-length). The results were compared to HET and WT injected with a tag-only vector (CTRL). It was found that the presence of a C-terminal-modified EAAT2 transgene (i) increased the level of native EAAT2 protein in striatal lysates and perisynaptic astrocyte processes, (ii) enhanced the glutamate uptake of transduced astrocytes, (iii) stimulated glutamate clearance at individual corticostriatal synapses, (iv) increased the glutamate uptake of striatal astrocytes and (iv) alleviated the mHTT-related hypokinesia (open field indicators of movement initiation). In contrast, over-expression of full-length EAAT2 neither facilitated glutamate uptake nor locomotion. Together, our results support the new hypothesis that preventing abnormal protein-protein interactions at the C-terminal of EAAT2 could eliminate the mHTT-related deficits in corticostriatal synaptic glutamate clearance and movement initiation

A novel Fontan Y-graft for interrupted inferior vena cava and azygos continuation

OBJECTIVES To evaluate the hemodynamicdynamic advantage of a new Fontan surgical template that is intended for complex single-ventricle patients with interrupted inferior vena cava-azygos and hemi-azygos continuation. The new technique has emerged from a comprehensive pre-surgical simulation campaign conducted to facilitate a balanced hepatic flow and somatic Fontan pathway growth after Kawashima procedure. METHODS For 9 patients, aged 2 to18 years, majority having poor preoperative oxygen saturation, a pre-surgical computational fluid dynamics customization is conducted. Both the traditional Fontan pathways and the proposed novel Y-graft templates are considered. Numerical model was validated against in vivo phase-contrast magnetic resonance imaging data and in vitro experiments. RESULTS The proposed template is selected and executed for 6 out of the 9 patients based on its predicted superior hemodynamic performance. Pre-surgical simulations performed for this cohort indicated that flow from the hepatic veins (HEP) do not reach to the desired lung. The novel Y-graft template, customized via a right- or left-sided displacement of the total cavopulmonary connection anastomosis location resulted a drastic increase in HEP flow to the desired lung. Orientation of HEP to azygos direct shunt is found to be important as it can alter the flow pattern from 38% in the caudally located direct shunt to 3% in the cranial configuration with significantly reversed flow. The postoperative measurements prove that oxygen saturation increased significantly (P-value = 0.00009) to normal levels in 1 year follow-up. CONCLUSIONS The new Y-graft template, if customized for the individual patient, is a viable alternative to the traditional surgical pathways. This template addresses the competing hemodynamic design factors of low physiological venous pressure, high postoperative oxygen saturation, low energy loss and balanced hepatic growth factor distribution possibly assuring adequate lung development. Date and number of IRB approval 25 October 2019, 280011928-604.01.01

ATP from synaptic terminals and astrocytes regulates NMDA receptors and synaptic plasticity through PSD-95 multi-protein complex

Recent studies highlighted the importance of astrocyte-secreted molecules, such as ATP, for the slow modulation of synaptic transmission in central neurones. Biophysical mechanisms underlying the impact of gliotransmitters on the strength of individual synapse remain, however, unclear. Here we show that purinergic P2X receptors can bring significant contribution to the signalling in the individual synaptic boutons. ATP released from astrocytes facilitates a recruitment of P2X receptors into excitatory synapses by Ca2+-dependent mechanism. P2X receptors, co-localized with NMDA receptors in the excitatory synapses, can be activated by ATP co-released with glutamate from pre-synaptic terminals and by glia-derived ATP. An activation of P2X receptors in turn leads to down-regulation of postsynaptic NMDA receptors via Ca2+-dependent de-phosphorylation and interaction with PSD-95 multi-protein complex. Genetic deletion of the PSD-95 or P2X4 receptors obliterated ATP-mediated down-regulation of NMDA receptors. Impairment of purinergic modulation of NMDA receptors in the PSD-95 mutants dramatically decreased the threshold of LTP induction and increased the net magnitude of LTP. Our findings show that synergistic action of glia- and neurone-derived ATP can pre-modulate efficacy of excitatory synapses and thereby can have an important role in the glia-neuron communications and brain meta-plasticity

Prevalence of Cryptosporidium Spp. in Camels and Involved People in Yazd Province, Iran

Background: Although infection of dromedary camels with Cryptosporidium spp. is rare in Iran, it is considered a zoonotic threat to the keepers and herders of camels. Thus we investigated the prevalence of Cryptosporidium in these two hosts in Yazd Province, a semi-arid region in center of Iran.Methods: This study was conducted during 4 seasons (winter 2008, summer 2009, winter 2009 and summer 2010). Fecal samples (n=200) were collected from live camels. Also, 100 abomasal mucosa and related fecal samples of the slaughtered camels were investigated. Stool samples from 100 individuals who were in persistent contact with camels were also obtained. After staining by modified Ziehl-Neelsen method, the prepared specimens were studied microscopically. Results were analyzed using SPSS 16.Results: The rate of infection in feces and abomasal mucosa of camels were 20.33% and 12%, respectively. In addition, simultaneous fecal and mucosal infection was detected in 3 cases in winter. Statistical analyses showed no significant relation between infection and age of camels, as well as their sex and the season. Cryptosporidiosis in people who were in long-term contact with camels was also investigated microscopically by obtaining stool samples of 100 individuals (50 in summers, 50 in winters), 24 of them being infected with Cryptosporidium spp. The rate of infection was higher in winter than summer (16/50 compared with 8/50).Conclusion: The prevalence of Cryptosporidium spp. in camels and involved humans in Yazd Province is relatively considerable and of public health importance

Assessment of laboratory test requests from three hospital emergency departments

Background: Laboratory requests can be justified and trimmed to what patients necessarily need. Reducing unnecessary laboratory requests can improve patient safety and both patient and provider satisfaction. Methods: In this study, the current status of laboratory requests was assessed in emergency departments of three multidisciplinary university hospitals based on ten most expensive and most requested laboratory tests and categorised by the ordering physicians and patients� chief complaints in a 6-month period. These hospitals had annual visits to the emergency departments of between 20, 000-72.000 patients. Results: The most requested tests were as follows: complete blood count, biochemistry profile including sodium, potassium, urea, and creatinine. Moreover, the cumulating most expensive tests were coagulation profile, cardiac troponin, C-reactive protein, erythrocyte sedimentation rate, liver enzymes, alkaline phosphatase, and blood gases corresponding to the frequency of requests in the emergency department. Among the services, other than emergency medicine, requesting laboratory work in the emergency departments, internal medicine, neurology and surgery services had requested more laboratory tests comprising 44.0, 24.4, and 20.8 of all specialties respectively. Moreover, the most frequent laboratory requests were from patients complaining of abdominal pain, chest pain, and penetrating or blunttraumatic injuries. Conclusions: It should be considered to rationally request the most frequent laboratory tests as many of them can be canceled by physicians and do not change the diagnosis, treatment, prognosis, and disposition in the emergency department. © 2021 The authors

Plasma levels of CTRP-3, CTRP-9 and apelin in women with multiple sclerosis

Background: Recently, adipocytokines have been shown to play a pivotal role in autoimmune and inflammatory-related disease. The purpose of this study was to compare the levels of CTRP3, CTRP9, adiponectin and apelin- in Multiple Sclerosis (MS) patients with healthy subjects and their relationship with clinical parameters and the levels of pro-inflammatory mediators. Methods: Plasma levels of CTRP3, CTRP9, apelin, TNF-α, hs-CRP, and adiponectin were evaluated in 24 healthy women and 26 women with relapsing�remitting MS using immunoassay methods. Results: The plasma apelin level of the MS patients was significantly lower than that of healthy controls. The concentration of TNF-α and adiponectin were significantly higher in MS patients compared to the healthy controls. Plasma CTRP3, CTRP9 and hs-CRP levels were not significantly different between the two groups. There was no correlation between these adipokines and inflammatory mediators. A statistically significant negative correlation was observed between plasma concentrations of apelin with expanded disability status scale (EDSS) scores and number of relapse. Conclusions: Our findings suggest that adipokines, particularly apelin and adiponectin, may contribute to the pathogenesis of MS and can be considered as a biomarker or as a therapeutic target for the treatment of this disease. © 2019 Elsevier B.V