Unstirred layer and kinetics of electrogenic glucose absorption in the human jejunum in situ.
Using an electrical technique we estimated the thickness of the unstirred layer in the human jejunum during kinetic studies of electrogenic glucose absorption. The unstirred layer in seven healthy volunteers (632 +/- 24 mum: mean +/- SEM) was significantly thicker than in 10 patients with active coeliac disease (442 +/- 23 mum) but not significantly different in seven patients who had responded to treatment by gluten withdrawal (585 +/- 49 mum). There were similar differences in the values of ;Apparent Km' for electrogenic glucose absorption between healthy control subjects (36 +/- 6 mM) active coeliac patients (11 +/- 1 mM) and treated coeliac patients (31 +/- 5 mM). The changes in PDmax however, showed a different pattern. The PDmax in the active coeliac group (6.8 +/- 0.7 mV) was lower than in controls (7.6 +/- 0.6 mV) but not significantly so, while the PDmax in the treated coeliac group (10.6 +/- 0.9 mV) was significantly higher than in both the active coeliac and control groups. It should be noted that both operational kinetic parameters obtained in the present study are much lower than those obtained previously (Read et al., 1976b) because of the use of siphonage. Analysis of the results using a computer simulation indicates that the reduction in Apparent Km in active coeliac disease can be caused by the interaction of the decreased maximal absorption rate for glucose (Jmax) with the attenuated unstirred layer. In these circumstances it is not necessary to postulate any change in the affinity of the transport mechanism for glucose (;Real Km'). It is remarkable that the disease process produces an Apparent Km which is much closer to the Real Km than that found in health.


The effect of gastric juice on interleukin-8 production by cystic fibrosis primary bronchial epithelial cells.
CF patients are often treated with proton pump inhibitors (PPIs) to reduce acidic gastro-esophageal reflux (GER) and bronchial aspiration of duodeno-gastric contents is common in CF. We have previously demonstrated that gastric juice (GJ) from patients "on" PPI can induce interleukin-8 (IL-8) production by bronchial epithelial cells in culture. We hypothesized that such effect would be more pronounced in CF patients known to have high inflammatory susceptibility. We aimed to evaluate the effect of GJ on IL-8 production by primary bronchial epithelial cells (PBEC), derived from a CF patient and a healthy subject. PBEC obtained from one donor (normal PBEC) and one receptor (CF-PBEC) for lung transplantation were stimulated with GJ from patients "off" and "on" PPI. IL-8 levels were measured in the supernatant. GJ from patients "on" PPI provoked a significant higher IL-8 production compared to GJ from patients "off" PPI, both in normal PBEC [462 (200-1468) vs. 11 (4-28) pg/ml, p=0.0001] as in CF-PBEC [1468 (841-2449) vs. 85 (26-131) pg/ml, p<0.0001]. Exposure of the cells to GJ "off" PPI and "on" PPI provoked significantly higher IL-8 production in the CF-PBEC compared to the normal PBEC ["off" PPI 85 (26-131) vs. 11 (4-28) pg/ml, p=0.01; "on" PPI 1468 (841-2449) vs. 462 (200-1468) pg/ml, p=0.01]. Filtration (0.20 μm) of the GJ "on" PPI, to eliminate large particles and bacterial sub-products, resulted in a significant decrease of IL-8 production. Patients with CF, treated with PPIs, have GJ with high pH and high endotoxin levels. These patients often have GER and bronchial aspiration. The aspirated material (GJ "on" PPI) has a significantly enhanced inflammatory effect on CF bronchial epithelial cells in culture. As chronic PPI treatment in CF may result in a paradoxically increased inflammatory effect in the airways, alternative anti-reflux therapies should be considered in CF.


Exploration of soil bacterial communities for their potential as bioresource.
Soil is a repository of diverse microorganisms, which has frequently been used to isolate and exploit microbes for industrial, environmental and agricultural applications. Knowledge about the structure and dynamics of bacterial communities in soil has been limited as only a small fraction of bacterial diversity is accessible to culture methods. Traditional enrichment techniques and the pure culture approach for microbiological studies have offered only a narrow portal for examining the soil microbial flora due to their limited selectivity. Therefore, the morphological and nutritional criteria used to describe bacterial community failed to provide a natural taxonomic order according to evolutionary relationship. Molecular methods under an emerging discipline of biology "molecular microbial ecology" are now helping in getting these constraints removed to some extent. Nucleic acid extraction from soil is the first crucial step in the application of most of the molecular techniques, which have largely been dominated by diverse variations of PCR. Due to its rapidity, sensitivity and specificity, PCR-based finger printing techniques have proved extremely useful in assessing the changes in microbial community structure. Such techniques can yield complex community profiles and can also provide useful phylogenetic information. Fluorescent in situ hybridization (FISH) can be used to evaluate the distribution and function of bacterial population in situ. DNA microarray techniques have also been developed and being frequently used for the evaluation of ecological role and phylogenetic affiliations of bacterial populations in the soil.


The long-term effects of transluminal balloon angioplasty for vasospasms after subarachnoid hemorrhage: analyses of cerebral blood flow and reactivity.
Transluminal balloon angioplasty (TBA) has come into wide use for management of symptomatic cerebral vasospasm after subarachnoid hemorrhage (SAH). The long-term effects of TBA in this clinical context on cerebral blood flow (CBF) and the functional properties of the arterial wall after aneurysmal SAH remain controversial. We therefore studied these effects. All patients underwent unilateral TBA. Xenon-enhanced computed tomography was performed for an average of 18 days after TBA to measure CBF and cerebrovascular reactivity (CVR). Cerebral blood flow and CVR were compared between the side of TBA and the contralateral side. Nineteen vascular territories were treated successfully with TBA in 12 patients. Angiographic improvement of vasospasm was demonstrated in all 12 patients, and 9 (75%) patients showed neurological improvement. After balloon angioplasty, global CBF was 35.1 +/- 8.2 mL/100 g per minute, with CBF on the side with TBA (37.8 +/- 10.3 mL/100 g per minute) being essentially the same as that on the other side (P = .0671, paired Student t test). Likewise, reactivity to acetazolamide did not differ significantly between sides (P = .0817). Transluminal balloon angioplasty increased proximal vessel diameters but showed no significant influence on CBF or vascular reactivity 3 weeks later. Benefits presumably were short term, but the procedure was clinically safe.


Accurate identification of polyadenylation sites from 3' end deep sequencing using a naive Bayes classifier.
3' end processing is important for transcription termination, mRNA stability and regulation of gene expression. To identify 3' ends, most techniques use an oligo-dT primer to construct deep sequencing libraries. However, this approach can lead to identification of artifactual polyadenylation sites due to internal priming in homopolymeric stretches of adenines. Although heuristic filters have been applied in these cases, they typically result in a high proportion of both false-positive and -negative classifications. Therefore, there is a need to develop improved algorithms to better identify mis-priming events in oligo-dT primed sequences. By analyzing sequence features flanking 3' ends derived from oligo-dT-based sequencing, we developed a naïve Bayes classifier to classify them as true or false/internally primed. The resulting algorithm is highly accurate, outperforms previous heuristic filters and facilitates identification of novel polyadenylation sites.


Switching on BTK-One Domain at a Time.
BTK kinase activity is controlled by multiple inhibitory domains, whose coordinated mechanism of action is poorly understood. In this issue of Structure,Joseph et al. (2017) use solution-based approaches to characterize conformational changes associated with the binding of each inhibitory tether, revealing a multi-step activation process and a previously unknown C-terminal autoinhibitory latch.


Synchronous video laryngoscopy and sonography of the larynx in children.
Unlike laryngoscopy, laryngeal sonography is easily applicable, well tolerated by young children and less exhausting for the examiner. The aim of this study was to determine if sonography can adequately visualize the laryngeal structures and vocal fold movements of children. Synchronic video laryngoscopy and sonography of the larynx were performed in 35 children (aged 2-8 years) without a laryngeal disease. 10 investigators at differing stages of otolaryngological training were tasked with identifying vocal fold movement and specific anatomical structures after seeing (1) the sonography-only video and (2) the sonography video with a laryngoscopy video overlay. The percentage of correct identification of the structures and the vocal cord movement with and without additional video-overlay was compared. A sonographical view of the larynx could be found and the following anatomical structures were identified: subcutaneous fat tissue, left and right sternohyoid and thyrohyoid muscles, thyroid cartilage, left and right vocal folds, left and right paraglottic spaces, left and right side arytenoid cartilages, and rima glottides. Concerning the evaluation of the videos amongst the investigators: they identified >80 % of vocal fold movement and the targeted anatomical structures except the arytenoid cartilages (only up to 63 %), having the sonography only. In combination with the laryngoscopy video-overlay, investigator detection rates significantly improved (>90 % of vocal fold movement, p < 0.001; >90 % in most anatomical structures, (p < 0,001). Laryngeal sonography is a helpful diagnostic tool to identify laryngeal structures and the movement of the vocal folds in children.


Characterization, enhancement and modelling of mannosylerythritol lipid production by fungal endophyte Ceriporia lacerate CHZJU.
The glycolipid biosurfactants mannosylerythritol lipids (MELs) attract great attention for their biodegradability, super emulsifying properties and versatile bioactivities. In this study, the MEL deriving from Ceriporia lacerate CHZJU was identified as MEL-A, and its critical micelle concentration and emulsifying activities were assessed. To examine the production of MELs from Ceriporia lacerate, a Plackett-Burman design and response surface methodology were used to optimize the culture nutrients. The optimal medium contains 1g/L yeast extract, 1.5g/L (NH4)2SO4, 0.5g/L KH2PO4, 0.04g/L CaCl2, 119.6mL/L soybean oil and 0.297g/L MnSO4. Subsequent verification revealed that the yield of MELs was 129.64±5.67g/L. Furthermore, an unstructured kinetic model was developed for mycelial growth, MEL production and substrate utilization. This work provides insight into Ceriporia lacerate CHZJU, a predominant fungus producing MEL-A. Optimization using response surface methodology enhanced the mannosylerythritol lipid recovery. Importantly, we developed fermentation kinetic modelling for mannosylerythritol lipid production.


A Low Dose Combination of Withaferin A and Caffeic Acid Phenethyl Ester Possesses Anti-Metastatic Potential In Vitro: Molecular Targets and Mechanisms.
Withaferin A (Wi-A) and Caffeic Acid Phenethyl Ester (CAPE) are the bioactive ingredients of Ashwagandha (Withania somnifera) and propolis, respectively. Both of these natural compounds have been shown to possess anticancer activity. In the present study, we recruited a low dose of each of these compounds and developed a combination that exhibited remarkably potent anti-migratory and anti-angiogenic activities. Extensive molecular analyses including a cDNA array and expression analyses of the specific gene targets demonstrated that such activities are mediated through their effect on cell adhesion/tight junction proteins (Claudins, E-cadherin), inhibition of canonical Wnt/β-catenin signaling pathways and the consequent downregulation of EMT-signaling proteins (Vimentin, MMPs, VEGF and VEGFR) that play a critical role in cancer metastasis. The data supported that this novel combination of Wi-A and CAPE (Wi-ACAPE, containing 0.5 µM of Wi-A and 10 µM of CAPE) may be recruited for the treatment of metastatic and aggressive cancers and, hence, warrant further evaluation by recruiting a variety of experimental and clinical metastatic models.


In vitro pharmacological evaluation of multitarget agents for thromboxane prostanoid receptor antagonism and COX-2 inhibition.
Patients with high cardiovascular risk due to ageing and/or comorbidity (diabetes, atherosclerosis) that require effective management of chronic pain may take advantage from new non-steroidal anti-inflammatory drugs (NSAIDs) that at clinical dosages may integrate the anti-inflammatory activity and reduced gastrointestinal side effects of selective cyclooxygenase-2 (COX-2) inhibitor (coxib) with a cardioprotective component involving antagonism of thromboxane A2 prostanoid (TP) receptor. New compounds were obtained modulating the structure of the most potent coxib, lumiracoxib, to obtain novel multitarget NSAIDs endowed with balanced coxib and TP receptor antagonist properties. Antagonist activity at TP receptor (pA2) was evaluated for all compounds in human platelets and in an heterologous expression system by measuring prevention of aggregation and Gq-dependent production of intracellular inositol phosphate induced by the stable thromboxane A2 (TXA2) agonist U46619. COX-1 and COX-2 inhibitory activities were assessed in human washed platelets and lympho-monocytes suspension, respectively. COX selectivity was determined from dose-response curves by calculating a ratio (COX-2/COX-1) of IC50 values. The tetrazole derivative 18 and the trifluoromethan sulfonamido-isoster 20 were the more active antagonists at TP receptor, preventing human platelet aggregation and intracellular signalling, with pA2 values statistically higher from that of lumiracoxib. Comparative data regarding COX-2/COX-1 selectivity showed that while compounds 18 and 7 were rather potent and selective COX-2 inhibitor, compound 20 was somehow less potent and selective for COX-2. These results indicate that compounds 18 and 20 are two novel combined TP receptor antagonists and COX-2 inhibitors characterized by a fairly balanced COX-2 inhibitor activity and TP receptor antagonism and that they may represent a first optimization of the original structure to improve their multitarget activity.


The prevalence and plasmid profile of non-typhoidal salmonellosis in children in Lagos metropolis, South-western Nigeria.
Non-typhoidal Salmonella is the causative agent of gastroenteritis, a food-borne and zoonotic infection which is a major cause of high morbidity and death among children under 5 years of age especially from resource poor settings like the developing countries. This study was carried out for 6 months to determine the prevalence and plasmid profile of non-typhoidal salmonellosis in children in Lagos metropolis. A total of 105 stool samples were collected from diarrheal children aged 3 months to 12 years and processed during this period. The isolates were identified using Selenite F Broth, Salmonella-Shigella Agar, Kligler Iron Agar, and Motility-indole-Urea medium, citrate and sugar utilization tests. A total number of 127 isolates were identified, 2 of which are Salmonella enteritidis (1.6%). The non-typhoidal Salmonellae were sensitive to ciprofloxacin, cetotaxime, streptomycin, cotrimxazole and tetracycline. Only one of the 2 isolates (50%) was sensitive to amoxillin and sulphonamide while none of them (0%) was sensitive to cefuroxime. The plasmid analysis of the isolates showed that they harboured no detectable plasmids; this suggests that the resistance was chromosomally mediated.


Noisy galvanic vestibular stimulation improves vestibular perception in bilateral vestibulopathy.
Patients with bilateral vestibulopathy (BVP) suffer from impaired vestibular motion perception that is linked to deficits in spatial memory and navigation. To examine the potential therapeutic effect of imperceptible noisy galvanic vestibular stimulation (nGVS) on impaired vestibular perceptual performance in BVP. In 11 patients with BVP (mean age: 54.0 ± 8.3 years, 7 females), we initially determined the nGVS intensity that optimally stabilizes balance during a static posturographic assessment. Subsequently, effects of optimal nGVS vs. sham stimulation on vestibular motion perception were examined in randomized order. Vestibular perceptual performance was determined as direction recognition thresholds for head-centered roll tilt motion on a 6DOF motion platform in the absence of any visual or auditory motion cues. For each patient, an nGVS intensity that optimally stabilized static balance compared to sham stimulation could be identified (mean 0.36 ± 0.16 mA). nGVS at optimal intensity resulted in lowered vestibular perceptual thresholds (0.94 ± 0.30 deg/s) compared to sham stimulation (1.67 ± 1.11 deg/s; p = 0.040). nGVS-induced improvements in vestibular perception were observed in 8 of 11 patients (73%) and were greater in patients with poorer perceptual performance during sham stimulation (R = - 0.791; p = 0.007). nGVS is effective in improving impaired vestibular motion perception in patients with BVP, in particular in those patients with poor baseline perceptual performance. Imperceptible vestibular noise stimulation might thus offer a non-invasive approach to target BVP-related impairments in spatial memory, orientation, and navigation.


The pharmacology of impulsive behaviour in rats IV: the effects of selective serotonergic agents on a paced fixed consecutive number schedule.
An impulsive cognitive style may affect behaviour in several different ways, including rapid decision making, intolerance of the delay of reward and a tendency to terminate chains of responses prematurely. It has been proposed to measure the last of these in rats using fixed consecutive number (FCN) schedules. The present study uses a modified version of the FCN procedure in which responding was paced by retracting the response lever for short periods between presses. In this way, the experimenter can control the maximum rate of responding. The procedure was made up of two components. In both, the schedule requirement was FCN 8, but in the Fast component lever presses were spaced by a minimum of 2.5 s and in the Slow component by a minimum of 5 s. Alterations in impulsivity were inferred from changes in the mean chain length and the distribution of chain lengths. The 5-HT1A agonist, 8-OH-DPAT (0.03-0.3 mg/kg), increased chain lengths within a narrow dose range, whereas the 5-HT1A antagonist, WAY 100 635 (0.03-0.3 mg/kg), reduced chain lengths. The 5-HT2 agonist, DOI (0.1-1.0 mg/kg), markedly reduced chain lengths, whereas the 5-HT2 antagonist, ritanserin (0.03-0.3 mg/kg), had no effect. The 5-HT1A/1b agonist, RU 24969 (0.03-0.3 mg/kg), reduced chain lengths. The 5-HT releaser, p-chloramphetamine (0.1-1.0 mg/kg), had a weak, biphasic effect, slightly reducing the number of short chains at the lowest dose tested and slightly increasing this number at the highest dose. Other drugs tested, citalopram (1.0-10.0 mg/kg), metergoline (0.3-3.0 mg/kg) and MDL-72222 (0.1-3.0 mg/kg), had no significant effects. These results suggest that stimulation of 5-HT1A receptors reduces impulsivity, whereas stimulation of 5-HT2 receptors increases it. These data are in agreement with previous results using the DRL-72 schedule, and indicate that there is no simple role for serotonin in the control of impulsivity.


Muscle glycogenosis.
This review describes clinical, biochemical and genetic features of the four inborn errors affecting muscle glycogen breakdown, namely deficiencies of phosphorylase, phosphorylase kinase, amylo-1,6-glucosidase and acid alpha-glucosidase. They are characterized by a wide spectrum of clinical manifestation, affecting age of onset, clinical features, progress of disease and tissue involvement. Biochemically, variability of all four enzyme deficiencies is evident in terms of differences in residual enzyme present in tissues, and in the presence or absence of enzyme protein. Genetic heterogeneity, which has been documented in each of the enzyme deficiencies, manifests itself in terms of the presence, absence, quantity or size of mRNA. In phosphorylase deficiency heterogeneity has also been documented at the DNA level. In acid maltase deficiency nine mutant phenotypes have been described affecting various stages of lysosomal enzyme processing.


Partial nephrectomy for a large renal lymphatic malformation in a child presenting with hypertension.
A 9-year-old girl had hypertension (systolic blood pressure of 125 mm Hg) noted at an annual well child visit. An ultrasound study demonstrated a large right renal cystic mass. A partial nephrectomy was performed. The surgical specimen was 9.7 × 9.4 × 6.4 cm and weighed 413.2 g. The tumor stained diffusely positive for smooth muscle actin and focally positive for factor VIII. Final histologic diagnosis was primary intrarenal lymphatic malformation. The case is unusual because of the presentation, size of the mass, as well as the therapeutic approach of partial nephrectomy.


Musculoskeletal imaging in the cat: what's normal? What's abnormal?
Despite the increasing availability of ultrasonography, computed tomography and magnetic resonance imaging (MRI), radiography remains the primary imaging modality for the assessment of feline musculoskeletal disease in practice. In many respects, having a more feline-focused approach to radiography will reward the clinician with better quality images and, hence, a greater likelihood of a correct diagnosis. Correct interpretation of radiographic films requires familiarity with some of the unique aspects of normal feline skeletal anatomy. For optimal patient management, the clinician also needs an appreciation of the distinct advantages of advanced imaging modalities in certain circumstances, in particular for head trauma patients. When considering what equipment is most suitable for musculoskeletal imaging, the small size of our feline patients means that image resolution is of primary importance. Choosing an x-ray system (film-screen or digital) that offers more detail, and selecting ultrasound probes of a higher frequency than would be used on larger canine patients, will go some way to improving the diagnostic yield of any feline imaging study. If there is the option of referral for MRI, and a choice between high-field and low-field MRI systems for the feline patient, a high-field system will always provide more detail. This review, drawing on the author's experience and established imaging and anatomical principles, aims to assist general veterinary practitioners in their approach to feline musculoskeletal imaging. It provides an overview of the imaging choices and techniques for different regions of the body, and gives examples of normal anatomy that is peculiar to the cat.


Biomolecular force measurements and the atomic force microscope.
The atomic force microscope (AFM) is a surface-sensitive instrument capable of imaging biological samples at nanometer resolution in all environments including liquids. The sensitivity of the AFM cantilever, to forces in the pico Newton range, has been exploited to measure breakaway forces between biomolecules and to measure folding-unfolding forces within single proteins. By attaching specific antibodies to cantilevers the simultaneous imaging of target antigens and identification of antigen-antibody interactions have been demonstrated.


A distinctive latitudinal trend of nitrogen isotope signature across urban forests in eastern China.
Rapid urbanization has greatly altered nitrogen (N) cycling from regional to global scales. Compared to natural forests, urban forests receive much more external N inputs with distinctive abundances of stable N isotope (δ15 N). However, the large-scale pattern of soil δ15 N and its imprint on plant δ15 N remain less well understood in urban forests. By collecting topsoil (0-20 cm) and leaf samples from urban forest patches in nine large cities across a north-south transect in eastern China, we analyzed the latitudinal trends of topsoil C:N ratio and δ15 N as well as the correlations between tree leaf δ15 N and topsoil δ15 N. We further explored the spatial variation of topsoil δ15 N explained by corresponding climatic, edaphic, vegetation-associated, and anthropogenic drivers. Our results showed a significant increase of topsoil C:N ratio towards higher latitudes, suggesting lower N availability at higher latitudes. Topsoil δ15 N also increased significantly at higher latitudes, being opposite to the latitudinal trend of soil N availability. The latitudinal trend of topsoil δ15 N was mainly explained by mean annual temperature, mean annual precipitation, and atmospheric deposition of both ammonium and nitrate. Consequently, tree leaf δ15 N showed significant positive correlations with topsoil δ15 N across all sampled plant species and functional types. Our findings reveal a distinctive latitudinal trend of δ15 N in urban forests and highlight an important role of anthropogenic N sources in shaping the large-scale pattern of urban forest 15 N signature.


[Lung cancer in Germany - the current state of management].
The care of lung cancer patients in Germany has not been systematically evaluated yet. The aim of this article is to give an overview on the current state of lung cancer care on the basis of existing data. In April and May 2010, a literature search was performed in order to collect relevant information concerning epidemiology as well as diagnostic, therapeutic (systemic therapy, radiotherapy, surgery, palliative therapy), and interdisciplinary structures in lung cancer treatment. The published database on lung cancer care in Germany is overall deficient. Treatment of lung cancer patients is mainly located in hospitals, particularly in chest clinics or specialised departments. The access of hospitals for an outpatient treatment as provided per § 116 b SGB V has not yet been realised in all German states. A systematic and prospective evaluation of lung cancer care is necessary in order to better allocate resources in the future.


Deprescribing anticholinergic medication in the community mental health setting: A quality improvement initiative.
Chronic anticholinergic medication (ACM) prescribing with antipsychotics when no longer clinically indicated can lead to serious side effects and adversely impact patient quality of life. Through modifications of previously described deprescribing strategies undertaken in an academic, schizophrenia clinic, this quality improvement (QI) project aimed to bolster ACM deprescription where clinically appropriate among patients with schizophrenia and other psychiatric disorders at a community mental health center. A multidisciplinary team involving clinical pharmacists and psychiatrists created web streaming video medical education accredited modules, supplemented by small groups sessions and case consultations, and provided these to psychiatrists and nurse practitioners at a community mental health center over a one-year period. Electronic medical record reports were also generated, highlighting patients who were receiving one of two ACM used in the clinic: benztropine and/or trihexyphenidyl; these were periodically distributed to support appropriate deprescription. Patient education infographic material focused on ACM were also created and deployed. One hundred and twenty-six patients were identified as receiving benztropine or trihexyphenidyl in March 2019. One hundred and six (84%) were on one or both of these medications for at least six months. The mean (±SD) age of the study sample was 53.4 (±12.6) years; a third of the sample was over 60 years. Thirty-seven patients, or 29.4%, had their ACM discontinued or the dosage reduced. Deprescription was not associated with age, sex, race, or diagnosis. Deprescription was not associated with antipsychotic polypharmacy, first versus second generation, or oral versus long acting preparation. These results suggest that deprescription of ACM in a community mental health center can occur with prescriber education and support. However, results from previous stages of this QI project, where much higher rates of deprescription were demonstrated, indicate the important benefit of more direct clinical pharmacist support and involvement in the process.


Relook on Mastoid Cavity Obliteration: A Prospective Study.
Canal wall down (CWD) mastoidectomy is the operation of choice for unsafe variety of chronic otitis media. But open mastoid cavity poses many problems. The solution of these problems is obliteration of mastoid cavity that is self-cleaning and easily maintained. In our study we aim to establish the effective technique for mastoid cavity obliteration in CWD mastoidectomy and review its efficacy in producing a dry, low maintenance cavity. This was a non-randomized longitudinal prospective study, performed over 2½ years in the department of ENT in a tertiary care hospital of Kolkata. Patients of chronic otitis media unsafe variety within the age group of 16-60 years were included in our study. Chronic otitis media unsafe variety with intratemporal or intracranial complications, and extensive cholesteatoma or granulation tissue that can't be cleared completely during operation were excluded. There was no statistical significance for hearing improvement between CWD mastoidectomy without obliteration and CWD mastoidectomy with obliteration. There was significant statistical significance for obliteration of cavity, epithelized cavity and dry cavity between CWD mastoidectomy without obliteration and CWD mastoidectomy with obliteration. Persistent discharge and granulation were significantly more in non-obliterated group. The time taken by the ear to become dry is much shorter after mastoid cavity obliteration. Moreover, lifelong aural toilet and dependence on an ENT surgeon is avoided. Inspite of all these, a few pre-conditions must be fulfilled before embarking on this type of surgery.


Nasolacrimal duct intubation in the treatment of congenital nasolacrimal duct obstruction in older children.
PurposeThe purpose of this study was to investigate the outcomes of nasolacrimal duct intubation in the primary treatment of congenital nasolacrimal duct obstruction (CNLDO) in children aged 7 years and older.MethodsThirty children aged ≥7 years who underwent primary unilateral nasolacrimal duct intubation because of CNLDO were enrolled in this study. CNLDO diagnosis was based on signs and symptoms including typical epiphora, increased tear meniscus, recurrent or persistent mucopurulent discharge, and an abnormal dye disappearance test (DDT). Surgical success was defined as complete resolution of previous signs and symptoms and DDT grade 0-1.ResultsThe mean age was 10.7±2.5 years (ranging from 7 to 15 years). Of the 30 patients, 18 were male and 12 were female. The mean follow-up period was 8.8±3.4 months (ranging from 6 to 16 months). The mean silicone tube removal time was 4.6±1.1 months (ranging from 3 to 6 months). The complete resolution of signs and symptoms with DDT grade 0-1 was observed in 22 of 30 cases (73.3%) during the follow-up period. The mean age of the patients with unsuccessful outcomes was 12.7±1.4 years, whereas that of the patients with successful outcomes was 10.0±2.4 years, and the difference was statistically significant (P=0.006). No serious intra- and/or post-operative complication was observed.ConclusionsThe current study demonstrates that nasolacrimal duct silicone intubation with intranasal endoscopic visualization has favorable outcomes as a primary treatment of persistent CNLDO in children aged 7 years and older. It can be used to reduce the need for dacryocystorhinostomy which is a more invasive procedure.


Age-related declines in distortion product otoacoustic emissions utilizing pure tone contralateral stimulation in CBA/CaJ mice.
One role of the medial olivocochlear (MOC) auditory efferent system is to suppress cochlear outer hair cell (OHC) responses when presented with a contralateral sound. Using distortion product otoacoustic emissions (DPOAEs), the effects of active changes in OHC responses due to the MOC as a function of age can be observed when contralateral stimulation with a pure tone is applied. Previous studies have shown that there are age-related declines of the MOC when broad band noise is presented to the contralateral ear. In this study, we measured age-related changes in CBA/CaJ mice by comparing DPOAE generation with and without a contralateral pure tone at three different frequencies (12, 22, and 37 kHz). Young (n = 16), middle (n = 10) and old-aged (n = 10) CBA mice were tested. DPOAE-grams were obtained using L1 = 65 and L2 = 50 dB SPL, F1/F2 = 1.25, using eight points per octave covering a frequency range from 5.6-44.8 kHz. The pure tone was presented contralaterally at 55 dB SPL. DPOAE-grams and ABR levels indicated age-related hearing loss in the old mice. In addition, there was an overall change in DPOAEs in the middle-aged and old groups relative to the young. Pure tone stimulation was not as effective as a suppressor compared to broadband noise. An increase in pure tone frequency from 12 to 22 kHz induced greater suppression of DPOAEs, but the 37 kHz was least effective. These results indicate that as the mouse ages, there are significant changes in the efficiency of the suppression mechanism as elicited by contralateral narrowband stimuli. These findings reinforce the idea that age-related changes in the MOC or the operating points of OHCs play a role in the progression of presbycusis - age-related hearing loss in mammals.


Mechanisms of early Recurrence in Intracranial Atherosclerotic Disease (MyRIAD): Rationale and design.
Intracranial atherosclerotic disease (ICAD) is the most common cause of ischemic stroke with the highest rate of recurrence, despite aggressive medical management. Diverse mechanisms may be responsible for ICAD-related cerebral ischemia, with potential therapeutic implications. Here we present the rationale, design and methods of the Mechanisms of Early Recurrence in Intracranial Atherosclerotic Disease (MyRIAD) study. The aim of MyRIAD is to determine the mechanisms of stroke in ICAD through physiologic imaging biomarkers that evaluate impaired antegrade flow, poor distal perfusion, abnormal vasoreactivity, artery to artery embolism, and their interaction. This is a prospective observational study of patients with recently symptomatic (<21 days) ICAD with 50-99% stenosis treated medically and monitored for up to 1 year. An estimated 110 participants are recruited at 10 sites to identify the association between the presence of each mechanism of ischemia and recurrent stroke. The primary outcome is ischemic stroke in the territory of the symptomatic artery. Secondary outcomes include new cerebral infarction on MRI at 6-8 weeks and recurrent TIA in the territory of the symptomatic artery. MyRIAD is positioned to define the role of specific mechanisms of recurrent ischemia in patients with symptomatic ICAD. This knowledge will allow the development and implementation of effective and specific treatments for this condition.


Precise alignment of peptidyl tRNA by the decoding center is essential for EF-G-dependent translocation.
Translocation is an essential step in the elongation cycle of the protein synthesis that allows for the continual incorporation of new amino acids to the growing polypeptide. Movement of mRNA and tRNAs within the ribosome is catalyzed by EF-G binding and GTP hydrolysis. The 30S subunit decoding center is crucial for the selection of the cognate tRNA. However, it is not clear whether the decoding center participates in translocation. We disrupted the interactions in the decoding center by mutating the universally conserved 16S rRNA bases G530, A1492, and A1493, and the effects of these mutations on translocation were studied. Our results show that point mutation of any of these 16S rRNA bases inhibits EF-G-dependent translocation. Furthermore, the mutant ribosomes showed increased puromycin reactivity in the pretranslocation complexes, indicating that the dynamic equilibrium of the peptidyl tRNA between the classical and hybrid-state configurations is influenced by contacts in the decoding center.


Improvement of mannitol production by Lactobacillus brevis mutant 3-A5 based on dual-stage pH control and fed-batch fermentations.
Lactobacillus brevis 3-A5 was isolated and expected to produce mannitol efficiently by regulating pH in batch and fed-batch fermentations. In 48 h batch fermentations with free and constant pH, the optimal pH for cell growth and mannitol production in the first 24 h of incubation was 5.5, whereas that for mannitol production in the second 24 h of incubation was 4.5. To achieve high cell density and mannitol yield simultaneously, a dual-stage pH control strategy was proposed based on the kinetic analysis of mannitol production. The pH value was controlled at 5.5 for the first 12 h of fermentation and subsequently shifted to 4.5 until the fermentation was completed. Under dual-stage pH control fermentation, a 103 g/L yield of mannitol with a volumetric production rate of 3.7 g/L/h was achieved after 28 h. The dual-stage pH control fed-batch fermentation strategy was further developed to improve mannitol yield, wherein the yield increased by 109 % to 215 g/L after 98 h of fermentation. This value is the highest yield of mannitol ever reported using L. brevis.


The Vascular Lab and the Interventional Radiologist.
A vascular laboratory (VL) can be defined as a place where physicians and technologists have a commitment to perform and interpret non-invasive testing with objective techniques that are most appropriate to the facility and the skills of the laboratory personnel. It provides the entry point for many patients as well as being the sight for diagnosis and follow up for patients with vascular disease for Interventional Radiologists (IRs). VLs may be run by a multi-specialty service, may be hospital-based, or may be private practice-based providing more of a concierge service. Resident education in the VL is essential and allows IRs to function on equal footing with other medical colleagues who see and treat similar patients from an early stage and throughout an IR's career. In this article we will discuss the IRs imperative connection to the VL.


Diquat increases cysteine proteinase inhibitors greatly in rat plasma and tissues.
Biochemical and gross pathological effects of diquat were studied with special attention to cysteine proteinase inhibitor level which was often increased in acute and chronic disorder. Diquat was fed continuously to rats at the dose of 1000 ppm in the diet. After 10 days, anorexia and severe diarrhea were observed but epistaxis and hypokinesia were not apparent. The rats were killed after feeding the diet for 13.5 days and plasma components such as acute phase reactant proteins and some vitamins which act as antioxidants were examined. The results showed that alpha-cysteine proteinase inhibitor (alpha-CPI) increased to 9-fold and vitamin C radical increased to 1.6-fold, whereas alpha 1 proteinase inhibitor (alpha 1-PI) decreased to 0.9-fold and vitamins C and E were the same as the control. Among three components of alpha-CPI, the T kininogen level in intoxicated rat plasma was about 20-fold, whereas the high molecular weight kininogen level was about 2-fold of the control. Diquat also enhanced the cysteine proteinase inhibitor (CPI) level to 20-fold in kidney and to 7- to 10-fold in the other organs. The large increment of T kininogen in these organs was also confirmed immunologically. The kidney showed a granular degeneration and its weight increased to 1.2-fold of control. The other organs showed neither gross pathological alteration nor weight change, compared with the control. The diquat distribution was highest in spleen and next highest in kidney among several organs. These results were compared with those caused by paraquat.


Effects of 10min vs. 20min passive rest after warm-up on 100m freestyle time-trial performance: A randomized crossover study.
The aim of this study was to compare the effect of 10min vs. 20min passive rest post warm-up on performance in a 100m freestyle time-trial. Randomized crossover. Eleven competitive male swimmers performed two experimental trials on different days, consisting of 100m freestyle time-trials following 10min or 20min passive rest after a standard 1200m warm-up. Performance (time-trial), biomechanical (stroke length, stroke frequency, stroke index, propelling efficiency), physiological (blood lactate concentrations, heart rate, core and tympanic temperature), and psychophysiological (perceived effort) variables were assessed during both trials. Time-trial performance was faster after 10min as opposed to 20min passive rest (58.41±1.99s vs. 59.06±1.86, p<0.01). This was supported by strong effect sizes (d=0.99) and the qualitative indication of "likely" positive effects. Heart rate before the time-trial was also higher after 10min passive rest (89±12bpm vs. 82±13bpm; p<0.01). Furthermore, net core temperature and oxygen uptake values before the time-trial were substantially lower after 20min passive rest. These data suggest that the 10min post warm-up passive rest enhances 100m freestyle performance when compared to a 20min period. An improvement that appears to be mediated by the combined effects of a shorter post warm-up period on core temperature, heart rate and oxygen uptake.


Prostatic abscess evaluated by serial computed tomography.
Prostatic abscess appears on computed tomography (CT) as multiple, well-demarcated fluid collections within the prostate gland and/or periprostatic tissues. Since prostatic abscess may not be differentiated from other prostatic disease on the basis of history and physical examination alone, CT can contribute significantly to establishing this diagnosis. Prostatic abscess can be an aggressive lesion within the pelvis and may rupture into the urethra, peritoneum, prevesical space, rectum, perineum, and ischiorectal fossa. By defining the extent of the disease, CT can guide selection of an optimal surgical drainage procedure. CT can be used effectively to monitor the treatment of prostatic abscess.


LncRNA NEAT1 Regulates the Development of Parkinson's Disease by Targeting AXIN1 Via Sponging miR-212-3p.
Long non-coding RNA (lncRNA) nuclear-enriched assembly transcript 1 (NEAT1) has been reported to be highly expressed in Parkinson's disease (PD). However, the mechanism of NEAT1 in PD progression has not been fully elucidated. 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine injection (MPTP) was used to construct PD mouse models in vivo, and 1-methyl-4-phenyl pyridine (MPP+) was used to build PD cell models in vitro. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to test the expression of NEAT1, microRNA (miR)-212-3p and axis inhibition protein 1 (AXIN1). The viability, apoptosis and inflammation of cells were determined using cell counting kit 8 (CCK8) assay, flow cytometry and enzyme-linked immunosorbent assay (ELISA), respectively. Then, the protein levels of apoptosis-related markers and AXIN1 were measured by western blot (WB) analysis. Furthermore, dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were performed to verify the interaction between miR-212-3p and NEAT1 or AXIN1. NEAT1 was upregulated in PD mouse models and cell models. Function experiments confirmed that NEAT1 knockdown could promote the viability, suppress the apoptosis and inflammation of MPP+-stimulated SK-N-SH cells to restrain PD progression. MiR-212-3p was downregulated in PD, and its inhibitor could reverse the suppression effect of NEAT1 knockdown on PD progression. Additionally, AXIN1 was a target of miR-212-3p, and its overexpression could invert the inhibition effect of miR-212-3p mimic on PD progression. Furthermore, AXIN1 expression was inhibited by NEAT1 silencing and promoted by NEAT1 overexpression, while these effect could be recovered by miR-212-3p inhibitor and mimic, respectively. Our results demonstrated that NEAT1 knockdown suppressed PD progression through regulating the miR-212-3p/AXIN1 pathway, indicating that NEAT1 might be a therapeutic target for neuroprotection in PD.


MiR-92a regulates brown adipocytes differentiation, mitochondrial oxidative respiration, and heat generation by targeting SMAD7.
Brown adipocytes are rich in mitochondria and linked to the body's blood fat levels and obesity. MiR-92a is negatively correlated with the activity of brown adipocytes. This study aimed to explore the mechanism of miR-92a on brown adipocytes. The expression of miR-92a in C2C12 cell was detected by a quantitative real-time-polymerase chain reaction (qRT-PCR). C2C12 cells were induced to brown adipocytes. The direct target gene of miR-92a was determined using the dual-luciferase reporter assay. Brown adipocytes were treated with isoprenaline (Iso) and transfected by miR-92a inhibitor and siSMAD7. The expression of heat-producing genes and adipose differentiation genes related to brown adipocytes were detected by qRT-PCR and Western blot analysis. The expression of SMAD7, p-SMAD2, and p-SMAD3 were detected using Western blot analysis. The mitochondrial content was measured by mitotracker fluorescent staining. MiR-92a inhibitor significantly decreased the expression of miR-92a in C2C12 cells. MiR-92a inhibitor could upregulate the expression of Ucp1, Cox7a1, Elovl3, Ppargc1α, PPARγ, and FABP4, and its effect on Ucp1 was increased after the treatment of isoprenaline. Moreover, miR-92a inhibitor increased mitochondrial content, oxygen consumption rate (OCR) and the expression of SMAD7 and suppressed the expressions of p-SMAD2 and p-SMAD3, whereas miR-92a directly targeted SMAD7 to exert its inhibitory effects. SiSMAD7 reversed the effects of the inhibitor on heat-producing genes, mitochondrial content, OCR and the expressions of SMAD7, p-SMAD2, and p-SMAD3 in brown adipocytes. Blocking miR-92a might promote brown adipocytes differentiation, mitochondrial oxidative respiration, and thermogenesis by targeting SMAD7 to inhibit the expressions of p-SMAD2 and p-SMAD3.


Surgical treatment of large and complicated hiatal hernias with the new resorbable mesh with hydrogel barrier (Phasix™ ST): a preliminary study.
Evaluation of the efficacy and safety of the new monofilament fully resorbable mesh with hydrogel barrier (Phasix™ ST), for large and complex hiatal hernia repair. Between December 2017 and December 2020, 60 patients with large or complicated hiatal hernia were treated (40 robotic and 20 laparoscopic procedures). The mesh was placed after primary closure of the hiatal defect, in an onlay fashion around the esophagus, followed by 360o fundoplication. Follow-up at 3, 6, 12, 18, 24 months from intervention included clinical evaluation and upper GI endoscopy. In cases of recurrence, radiologic survey and manometry were utilized. There were no conversions to open repair or significant postoperative incidents. Over a median follow-up of 21 months (range 3-36), no recurrences or mesh related complications were observed. From our early experience, Phasix™ ST mesh seems to be safe and effective for the reinforcement of crural defects in large and complex hiatal hernia.


Stable high expression of human gamma-aminobutyric acidA receptors composed of alpha and beta subunits.
Multiple classes of pharmacological agents including benzodiazepines, cage convulsants like t-butylbicyclophosphorothionate (TBPS), barbiturates and neuroactive steroids allosterically modulate the gamma-aminobutyric acidA receptor-chloride ionophore complex (GRC). The function of benzodiazepines requires a GRC comprised of alpha, beta and gamma subunits, while TBPS, barbiturates and neuroactive steroids will allosterically modulate GRCs comprised of only alpha and beta subunits. Binary alpha beta complexes are still hypothesized to be expressed in the mammalian brain particularly during development and could contribute to the pharmacological action of neuroactive steroids and barbiturates. In order to examine binary alpha beta complexes we report here the establishment of stable cell lines that express high levels of human GABAA receptors comprised of alpha 1 beta 1, alpha 2 beta 1 and alpha 3 beta 1 subunit combinations. The apparent potencies for allosteric modulation of [35S]TBPS for most naturally occurring neuroactive steroids for the binary subunit combinations was similar to that of the gamma-containing subunit combinations. Also discussed is the usefulness of these cell lines for the biophysical analysis of the GABAA receptor stoichiometry.


Italian multicentre study of carpal tunnel syndrome: clinical-neurophysiological picture and diagnostic pathway in 461 patients and differences between the populations enrolled in the northern, central and southern centres. Italian CTS Study Group.
The economic trend in our country, as in other countries, now requires us to pay more attention to the social and economic aspects of health. Therefore, it is important to evaluate the diagnostic pathway and the socioeconomic aspects of pathologies. The Italian CTS Study Group performed a wide multicentre and multidimensional study (through a patient-, physician- and neurophysiologically oriented evaluation) on 461 idiopathic CTS patients to evaluate the clinical-neurophysiological picture and the diagnostic pathway. The whole population and the differences between the populations enrolled in the neurophysiological centres of the northern, central and southern parts of Italy were analysed. This study provided new and interesting information. As concerns the whole population, we observed different diagnostic pathways in relation to the level of schooling. As concerns the comparison of the populations, the southern centres have a population of CTS patients with higher frequency of hand stress and more severe nerve impairment.


Influence of preovulatory estradiol treatment on the maintenance of pregnancy in beef cattle receiving in vivo produced embryos.
This experiment was designed to determine the role of preovulatory estradiol in pregnancy retention after embryo transfer (ET). Cows were synchronized with the 7-d CO-Synch + CIDR® protocol. On d0 (d-2 =CIDR® removal), cows were grouped by estrual status (estrual [Positive Control] and nonestrual), and nonestrual cows were administered Gonadotropin Releasing Hormone (GnRH) and randomly assigned to either no treatment (Negative Control) or Estradiol (0.1 mg estradiol 17-β IM). All cows received an embryo on d7. Pregnancy status was retrospectively classified on d56, 30, 24, and 19 by either ultrasonography, plasma pregnancy-associated glycoproteins analysis (PAGs), expression of interferon-stimulated genes, plasma progesterone (P4) concentrations, or a combination of the factors. There was no difference in estradiol concentrations on day 0 h 0 (P > 0.16). At day 0 h 2, Estradiol cows (15.7 ± 0.25 pg/mL) had elevated (P < 0.001) estradiol compared with Positive Controls (3.4 ± 0.26 pg/mL) or Negative Controls (4.3 ± 0.25 pg/mL). On d19, pregnancy rates did not differ (P = 0.14) among treatments. On d24, Positive Controls (47%) had greater (P < 0.01) pregnancy rates than Negative Controls (32%); Estradiol cows were intermediate (40%). There was no difference (P = 0.38) in pregnancy rates between Positive Control (41%) and Estradiol (36%) cows on d30, but Negative Control (27%) cows had (P = 0.01) or tended (P = 0.08) to have decreased pregnancy rates, respectively. Thus, preovulatory estradiol may elicit an effect on early uterine attachment or alter histotroph components, consequently improving pregnancy maintenance through d30.


Heat-induced interaction between egg white protein and wheat gluten.
Some wheat-based food systems, such as cakes and cookies, include mixtures of gluten and egg white protein (EWP) and are processed under heating conditions. Changes in these proteins during processing can affect the quality of the end product. This study investigated protein polymerization during heating of (mixtures of) wheat gluten and EWP. Chemical changes were studied by size-exclusion high performance liquid chromatography (SE-HPLC), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), thiol (SH) measurement and Fourier transform infrared (FTIR). During heating, protein polymerization was observed in the mixtures of gluten, glutenin, gliadin and EWP according to SE-HPLC profiles and results of SDS-extractable protein. The results of SDS-PAGE profiles of different proteins were in accordance with SE-HPLC. The number of SH groups in the majority of proteins showed a significant decrease, implying that disulfide (SS) bonds contributed to the extractability loss. In addition, changes of secondary structure tested by FTIR indicated protein aggregation.


Cdk inhibitor ste9p/srw1p is involved in response to protein synthesis inhibition in fission yeast.
It remains unknown whether the cell cycle system responds properly to protein synthesis inhibition. In this paper I report finding in Schizosaccharomyces pombe that partially deleted elongation factor 3 genes rescue various mitotic catastrophe mutants depending on deltaste9 in a dominant-negative manner. In response to protein synthesis inhibitors, deltaste9 and some other mutants delay halting the cell cycle at G2-M and the combined cdc2-M26 deltaste9 mutant greatly loses viability. It is suggested that cell cycle be positively controlled in an ste9-dependent manner before essential factors for viability and other important functions are exhausted when protein synthesis is inhibited.


Effect of a desk attachment board on posture and muscle activity in women during computer work.
Working at a computer is part of a large number of jobs and has been associated with upper extremity musculoskeletal disorders and back pain. The study evaluated the effects of a board attachment on upper extremity and back. The findings are mixed in that the board may have a positive effect in preventing back pain, but may be detrimental to upper extremities. Effect of a desk attachment board on upper extremity and trunk posture, and muscle activity was assessed in women video display terminal users. Participants completed a standard 20-min computer task under two conditions: 1) using a standard desk; 2) using a desk attachment board designed to support the forearms. Bilateral electromyography of the trapezius, multifidus and longissimus muscles and the right anterior deltoid and forearm extensor muscles was recorded. 3-D trunk and upper extremity posture was monitored. Participants were tested before and after 2 weeks of familiarisation with the board in their workplace. Perceived tension and discomfort were recorded before and after use of the board. Use of the board tended to increase muscle activity in the right trapezius and forearm extensor and to decrease muscle activity in the back. Perceived tension in the low back decreased slightly with the board. The board may be useful in reducing tension in the low back during computer work, but may adversely affect the upper extremities.


Waist circumference, visceral abdominal fat thickness and three components of metabolic syndrome.
The relationships between the waist circumference (WC), visceral adipose tissue (VAT) thickness and three components of metabolic syndrome (MetS) were explored to verify which of the obesity indices might be superior. A cross-sectional study was conducted of 1256 subjects (840 males and 416 females) aged 26-89 years, who were individuals undergoing intensive health checkup. The three components of MetS examined were high blood pressure, glucose intolerance and dyslipidemia. Receiver-operating characteristic (ROC) curve analysis and multiple logistic regression analysis were used for the analyses. The mean values of the WC and VAT thickness were significantly higher in the subject group positive for each of the metabolic syndrome components than in the subject group that was negative for all the components (p<0.001). ROC curve analysis showed that the WC showed an advantage over the VAT thickness only for predicting high blood pressure in men. Logistic regression analysis revealed increase of the odds ratios of the WC for glucose intolerance (p<0.001), high blood pressure (p<0.001) and dyslipidemia (p<0.01) in men. In contrast, the odds ratio of the VAT thickness was significantly increased only for dyslipidemia (p<0.01) in men. In women, the odds ratios of the WC for glucose intolerance (p<0.01) and dyslipidemia (p<0.001) were significantly increased, and odds ratio of the VAT thickness for high blood pressure (p<0.01) was significantly increased. This survey presented an advantage of WC over VAT thickness as an obesity index for identifying all the three components of metabolic syndrome, although sex differences in the study outcomes were found.


The Use of a "Double-Triple Barrel" Technique during Minimally Invasive Multilevel Tubular Laminectomy. A Technical Note.
This report presents a modified use of multiple tubular retractors (that limits issues related to the sequential redocking of tubular systems) that may prove helpful in cases where a minimally invasive approach is utilized for the treatment of multilevel lumbar stenosis.


The diagnostic values of red flags in pediatric patients with headache.
Headache is a common complaint in childhood and adolescence. Differentiating benign primary headaches from ominous secondary headaches is often difficult. Clinicians usually seek red flags to determine the need for neuroimaging. We aimed to evaluate the diagnostic values of red flags in pediatric headaches. We retrospectively reviewed the medical records of 1510 pediatric patients (1470 with primary headache, 40 with secondary headache) presenting with headache and underwent neuroimaging from two centers between March 2010 and December 2019. The secondary-headache group exhibited significantly higher frequencies of abnormal neurologic signs/symptoms (40.0% vs 6.8%, p < 0.001), Valsalva maneuver/exercise-induced headache (15.0% vs 4.9%, p = 0.004), headache with vomiting (35.0% vs 17.9%, p = 0.006), and onset under age 6 (25.0% vs 10.3%, p = 0.003) than the primary-headache group, with the following positive likelihood ratio (PLR): 5.88, 3.06, 1.96, and 2.42, respectively. The sensitivity values were as follows: abnormal neurologic signs/symptoms (16/40, 40.0%), headache with vomiting (14/40, 35.0%), onset under age 6 (10/40, 25.0%), and Valsalva maneuver/exercise-induced headache (6/40, 15.0%). The overall sensitivity for ominous secondary headaches requiring surgical treatment was 86.2% (25/29). Certain red flags, including abnormal neurologic signs/symptoms, Valsalva maneuver/exercise-induced headache, headache with vomiting, and onset under age 6, were more prevalent in the secondary-headache group; nonetheless, their sensitivity values and PLR were relatively low. Notwithstanding, considering these red flags' high overall sensitivity for ominous secondary headaches, neuroimaging in patients presenting these red flags should rely on careful follow-up of symptom progression.


Rho GTPases in PC-3 prostate cancer cell morphology, invasion and tumor cell diapedesis.
The Rho GTPases comprise one of the eight subfamilies of the Ras superfamily of monomeric GTP-binding proteins and are involved in cytoskeletal organization. Previously, using a dominant negative construct, we demonstrated a role for RhoC GTPase in conferring invasive capabilities to PC-3 human prostate cancer cells. Further, we demonstrated that inactivation of RhoC led to morphological changes commensurate with epithelial to mesenchymal transition (EMT) and was accompanied by increased random, linear motility and decreased directed migration and invasion. EMT was related positively to sustained expression and activity of Rac GTPase. In the current study we analyze the individual roles of RhoA, RhoC and Rac1 GTPases in PC-3 cell directed migration, invasion and tumor cell diapedesis across a human bone marrow endothelial cell layer in vitro. Use of specific shRNA directed against RhoA, RhoC or Rac1 GTPases demonstrated a role for each protein in maintaining cell morphology. Furthermore, we demonstrate that RhoC expression and activation is required for directed migration and invasion, while Rac1 expression and activation is required for tumor cell diapedesis. Inhibition of RhoA expression produced a slight increase in invasion and tumor cell diapedesis. Individual Rho GTPases are required for critical aspects of migration, invasion and tumor cell diapedesis. These data suggest that coordinated activation of individual Rho proteins is required for cells to successfully complete the extravasation process; a key step in distant metastasis.


Biological evaluation and osteogenic potential of polyhydroxybutyrate-keratin/Al2O3 electrospun nanocomposite scaffold: A novel bone regeneration construct.
In this study, the effect of alumina nanowire on the physical and biological properties of polyhydroxybutyrate-keratin (PHB-K) electrospun scaffold was investigated. First, PHB-K/alumina nanowire nanocomposite scaffolds were made with an optimal concentration of 3 wt% alumina nanowire by using the electrospinning method. The samples were examined in terms of morphology, porosity, tensile strength, contact angle, biodegradability, bioactivity, cell viability, ALP activity, mineralization ability, and gene expression. The nanocomposite scaffold provided a porosity of >80 % and a tensile strength of about 6.72 MPa, which were noticeable for an electrospun scaffold. AFM images showed an increase in surface roughness with the presence of alumina nanowires. This led to an improvement in the degradation rate and bioactivity of PHB-K/alumina nanowire scaffolds. The viability of mesenchymal cells, alkaline phosphatase secretion, and mineralization significantly increased with the presence of alumina nanowire compared to PHB and PHB-K scaffolds. In addition, the expression level of collagen I, osteocalcin, and RUNX2 genes in nanocomposite scaffolds increased significantly compared to other groups. In general, this nanocomposite scaffold could be a novel and interesting construct for osteogenic induction in bone tissue engineering.


Unique effects of apathy and depression signs on cognition and function in amnestic mild cognitive impairment.
Apathy and depression are the most common neuropsychiatric features in mild cognitive impairment (MCI). These syndromes have been linked to cognitive and functional decline. This study aimed to examine their unique relationships with specific cognitive abilities and daily functioning among MCI patients with memory impairment (amnestic MCI). Data from 90 individuals referred for outpatient clinical neuropsychological evaluation and subsequently characterized as having amnestic MCI were obtained from a memory disorders center. MCI was defined using current NIA-Alzheimer's Association criteria. Variables included caregiver-reported apathy (Frontal Systems Behavior Scale) and depression (Memory and Behavioral Problems Checklist), scores on neuropsychological measures, and the Lawton-Brody Instrumental Activities of Daily Living Scale. Data were analyzed with multiple regressions controlling for age and education. Depression was independently associated with composite scores of executive functioning. Neither apathy nor depression was associated with composite scores of attention, memory, or language. With regard to individual measures of executive functioning, depression was independently associated with poorer set-shifting (i.e., Trails B). Apathy was independently associated with poorer letter fluency (i.e., FAS). Apathy, but not depression, was associated with greater functional impairment. Apathy and depression are associated with different aspects of executive functioning in amnestic MCI, which may reflect differing patterns of frontal lobe pathology. Apathy exerts a greater impact on daily functioning than depression and may therefore increase reliance on caregivers. Results support the separability of apathy and depression in amnestic MCI. Clinicians should differentiate these neuropsychiatric states during assessment and treatment.


Chronic cold exposure increases skeletal muscle oxidative structure and function in Monodelphis domestica, a marsupial lacking brown adipose tissue.
Monodelphis domestica (Marsupialia: Didelphidae) was used as a model animal to investigate and compare muscle adaptation to exercise training and cold exposure. The experimental treatment consisted of four groups of animals: either warm or cold acclimation temperature and with or without endurance exercise training. Maximal aerobic capacity during a running VO2max test in the warm-exercised or cold-exposed (with or without exercise) groups was about 130 mL O(2)/kg/min, significantly higher than the warm-acclimated controls at 113.5 mL O(2)/kg/min. Similarly, during an acute cold challenge (VO2summit), maximal aerobic capacity was higher in these three experimental groups at approximately 95 mL O(2)/kg/min compared with 80.4 mL O(2)/kg/min in warm-acclimated controls. Respiratory exchange ratio was significantly lower (0.89-0.68), whereas relative heart mass (0.52%-0.73%) and whole-body muscle mitochondrial volume density (2.59 to 3.04 cm(3)) were significantly higher following cold exposure. Chronic cold exposure was a stronger stimulus than endurance exercise training for tissue-specific adaptations. Although chronic cold exposure and endurance exercise are distinct challenges, physiological adaptations to each overlap such that the capacities for aerobic performance in response to both cold exposure and running are increased by either or both treatments.


An Emotional Go/No-Go fMRI study in adolescents with depressive symptoms following concussion.
Following concussion, adolescents may experience both poor inhibitory control and increased depressive symptoms. fMRI research suggests that adolescents with major depressive disorder have abnormal physiological responses in the frontostriatal pathway, and exhibit poorer inhibitory control in the presence of negatively-aroused images. The scarcity of information surrounding depression following concussion in adolescents makes it difficult to identify patients at risk of depression after injury. This is the first study to examine neural activity patterns in adolescents with post-concussive depressive symptoms. To explore the effect of depressive symptoms on inhibitory control in adolescents with concussion in the presence of emotional stimuli using fMRI. Using a prospective cohort design, 30 adolescents diagnosed with concussion between 10 and 17years were recruited. The Children's Depression Inventory questionnaire was used to divide participants into two groups: average or elevated levels of depressive symptoms. Participants completed an Emotional Go/No-Go task involving angry or neutral faces in a 3Telsa MRI scanner. Eleven participants had elevated depressive symptoms, of which 72% were hit in the occipital region of the head at the time of injury. fMRI results from the Emotional Go/No-Go task revealed activity patterns in the overall sample. Faces activated regions associated with both facial and cognitive processing. However, frontal regions that are usually associated with inhibitory control were not activated. Adolescents with elevated levels of depressive symptoms engaged more frontal lobe regions during the task than the average group. They also showed a trend towards worse symptoms following MRI scanning. Adolescents with elevated depressive symptoms engaged brain regions subserving evaluative processing of social interactions. This finding provides insight into the role the environment plays in contributing to the cognitive demands placed on adolescents recovering from concussion.


Paget's disease of bone in South Africa.
A survey of Paget's disease of bone in Johannesburg South Africa, has revealed a prevalence of 2.4% in whites aged 55 years and over, which is similar to some centres in Europe; it has also revealed an unexpectedly high prevalence of 1.3% among blacks. These findings are discussed in relation to recent concepts of the pathogenesis of Paget's disease.


Degradation of human secretory immunoglobulin A by Blastocystis.
Microbial immunoglobulin A (IgA) proteases cleave human secretory IgA, promoting the mucosal adhesion of pathogens. To investigate if the enteric protozoan Blastocystis degrades human secretory IgA, cell lysate and conditioned medium from two species were exposed to immunoglobulin A. Secretory IgA was cleaved by both cell lysate and conditioned medium with mainly cysteine proteinase activity in B. hominis B isolate and aspartic proteinase activity in B. ratii WR1 isolate. These findings suggest that Blastocystis proteases may play a role in parasite survival in vivo.


Redesigning emergency department patient flows: application of Lean Thinking to health care.
To describe in some detail the methods used and outcome of an application of concepts from Lean Thinking in establishing streams for patient flows in a teaching general hospital ED. Detailed understanding was gained through process mapping with staff followed by the identification of value streams (those patients likely to be discharged from the ED, those who were likely to be admitted) and the implementation of a process of seeing those patients that minimized complex queuing in the ED. Streaming had a significant impact on waiting times and total durations of stay in the ED. There was a general flattening of the waiting time across all groups. A slight increase in wait for Triage categories 2 and 3 patients was offset by reductions in wait for Triage category 4 patients. All groups of patients spent significantly less overall time in the department and the average number of patients in the ED at any time decreased. There was a significant reduction in number of patients who do not wait and a slight decrease in access block. The streaming of patients into groups of patients cared for by a specific team of doctors and nurses, and the minimizing of complex queues in this ED by altering the practices in relation to the function of the Australasian Triage Scale improved patient flow, thereby decreasing potential for overcrowding.


Defining Content for a Competency-based (CanMEDS) Postgraduate Curriculum in Ambulatory Care: a Delphi Study.
Ambulatory training in internal medicine has been noted to be dysfunctional and inadequate. In this study, we developed a set of competency-based outcomes specific to ambulatory care to guide the design, implementation and evaluation of instructional events to ensure that societal needs are addressed. In 2007 a Delphi technique was used to reach consensus and define the priorities for competency-based training in ambulatory care for internal medicine residents. Four groups of stakeholders in Canada participated: program directors, members of the Canadian Society of Internal Medicine, recent graduates, and residents. Two rounds of the Delphi process were required to reach consensus on a set of sixty competency-based educational objectives in ambulatory care that were classified under the CanMEDS roles. The inclusion of recent graduates in this study resulted in the addition of non-clinical topics that would have otherwise been missed, falling under roles historically viewed as being challenging to teach and evaluate (Manager, Health Advocate). This study is the first time a Delphi-process has been used to define the priorities for ambulatory care training in internal medicine under a competency-based framework. The resulting compendium of competency-based objectives provides a foundation from which educators can design, evaluate and modify existing training experiences.


Genetics researchers' and IRB professionals' attitudes toward genetic research review: a comparative analysis.
Genetic research involving human participants can pose challenging questions related to ethical and regulatory standards for research oversight. However, few empirical studies describe how genetic researchers and institutional review board (IRB) professionals conceptualize ethical issues in genetic research or where common ground might exist. Parallel online surveys collected information from human genetic researchers (n = 351) and IRB professionals (n = 208) regarding their views about human participant oversight for genetic protocols. A range of opinions were observed within groups on most issues. In both groups, a minority thought it likely that people would be harmed by participation in genetic research or identified from coded genetic data. A majority of both groups agreed that reconsent should be required for four of the six scenarios presented. Statistically significant differences were observed between groups on some issues, with more genetic researcher respondents trusting the confidentiality of coded data, fewer expecting harms from reidentification, and fewer considering reconsent necessary in certain scenarios. The range of views observed within and between IRB and genetic researcher groups highlights the complexity and unsettled nature of many ethical issues in genome research. Our findings also identify areas where researcher and IRB views diverge and areas of common ground.


NAD-dependent DNA-binding activity of the bifunctional NadR regulator of Salmonella typhimurium.
NadR is a 45-kDa bifunctional regulator protein. In vivo genetic studies indicate that NadR represses three genes involved in the biosynthesis of NAD. It also participates with an integral membrane protein (PnuC) in the import of nicotinamide mononucleotide, an NAD precursor. NadR was overexpressed and purified as a His-tagged fusion in order to study its DNA-binding properties. The protein bound to DNA fragments containing NAD box consensus sequences. NAD proved to be the relevant in vivo corepressor, but full NAD dependence of repressor activity required nucleotide triphosphates. DNA footprint analysis and gel shift assays suggest that NadR binds as a multimer to adjacent NAD boxes. The DNA-repressor complex would sequester a potential RNA polymerase binding site and thereby decrease expression of the nad regulon.


Squalene isolated from Schizochytrium mangrovei is a peroxisome proliferator-activated receptor-α agonist that regulates lipid metabolism in HepG2 cells.
Peroxisome proliferator-activated receptor-alpha (PPARα) is a nuclear receptor that has critical roles in the treatment of atherosclerosis, hyperlipidemia and hepatic steatosis. Squalene is a novel nature PPARα agonist, identified from reporter gene assay and qPCR analysis. Cultured hepatocytes stimulated with squalene exhibited significantly decreased cellular triacylglycerols and cholesterol concentrations, while cellular uptake of fatty acids was increased. Quantitative PCR analysis revealed that expression of genes related to fatty acid uptake, fatty acid oxidation, ketogenesis and reverse cholesterol transport metabolism were upregulated, while that of genes related to fatty acid synthesis were suppressed in cell treated with squalene. Squalene is hypolipidemic by activation of PPARα via a ligand-mediated mechanism that regulates the expression of lipid metabolism genes in hepatocytes.


CT and MRI findings of cyclosporine-related encephalopathy and hypertensive encephalopathy.
We present the MRI and CT findings of one child with cyclosporine-related encephalopathy, and one child with hypertensive encephalopathy following cyclosporine-related encephalopathy. The imaging findings were shown well on T2-weighted and fluid-attenuated inversion recovery (FLAIR) MR images. Cyclosporine-related encephalopathy was distributed predominantly in the posterior white matter. Hypertensive encephalopathy showed similar changes of CT attenuation, but with wider distribution. These two disorders seem to have the same pathogenesis.


Laparoscopic Davydov correction of a failed gracilis flap neovagina in a patient with Mayer-Rokitansky-Kuster-Hauser syndrome with a pelvic kidney.
The laparoscopic Davydov procedure is a surgical technique for creation of a neovagina in patients with Mayer-Rokitansky-Kuster-Hauser syndrome (MRKH) or vaginal agenesis. Herein we report its use in creating a neovagina after failure of a previous surgical attempt in a patient with a pelvic kidney, which has not been described to date. The patient, a 28-year-old woman with MRKH in whom creation of a neovagina using bilateral gracilis flaps had been attempted, was unable to have intercourse because of a shortened and scarred vagina. We successfully used the laparoscopic Davydov procedure to create a functional neovagina despite the previous surgery and the presence of a pelvic kidney. The Davydov procedure is an option for use in patients with MRKH with history of unsuccessful neovaginal surgery and can be performed in the presence of a pelvic kidney.


Potentiation of tumor necrosis factor-mediated apoptosis of oral squamous cell carcinoma cells by adenovirus-mediated gene transfer of NF-kappaB inhibitor.
Oral squamous cell carcinoma (SCC) is a malignant tumor which is often resistant to cancer-therapy-mediated apoptosis. The stress-responsive transcription factor nuclear factor kappa B (NF-kappaB), which has been found to be associated with SCC development, plays an essential role in the suppression of tumor necrosis factor (TNF)-mediated apoptosis. Here, we report that an adenovirus-mediated gene transfer of NF-kappaB inhibitor, super-repressor I kappa B alpha (Adv-SR-IkappaBalpha), blocked TNF-induced NF-kappaB activation and sensitized oral SCC cells to TNF killing. Additionally, we found that the inhibition of NFkappaB by Adv-SR-IkappaBalpha enhanced TNF-mediated caspase-8 and -3 activation. These results suggest that NF-kappaB activation is a general mechanism by which oral squamous carcinoma cells are resistant to TNF killing and provide a molecular basis for gene therapy of oral cancer by IkappaBalpha gene transfer in vivo.


Effects of the COVID-19 pandemic on obstetrics and gynecology residency training in Turkey
This study aimed to evaluate the effect of the coronavirus disease-19 (COVID-19) pandemic on obstetrics and gynecology residency in Turkey. A 40-item questionnaire was prepared by the European Network of Trainees in Obstetrics and Gynecology. The survey included four parts, namely, workload, training aspects, trainees' health and safety, and women's and maternal health, conducted between April 2020 and September 2020. The submission of the questionnaire was voluntary. A total of 103 trainees from 28 cities responded to the survey. The mean duration of training was 2.5 years, and first- to fifth-year residents were included. In this study, 66 trainees (65.3%) were deployed in COVID-19 units, and the number of working hours was 84 hours per week. Moreover, 67% of the trainees reported insufficient outpatient clinic experience to meet education targets. Almost all trainees (101 of 103) trainees reported that the number of surgeries and/or elective surgeries decreased or were canceled. In addition, 63% and 68% of the trainees reported that their surgical skills were hindered by the reduced number of surgeries and dissatisfaction by not achieving a sufficient number of surgeries, respectively. Overall, 71% (n=73) were worried about their training. Only 45 trainees (43.6%) have raised their concerns to their program coordinators. Trainees in Turkey experienced a decrease in the workload during the first COVID-19 wave by 62% (n=64). In average, trainees worked nearly 30 h less than their usual workweek. Only 5% of the trainees (n=5) worked from home. Trainees claimed to have used sufficient personal protective equipment, and 66% (n=68) could keep their social distance in the hospital. The availability of health care was different between departments, and the family planning and reproductive medicine departments were the most affected. The obstetrics and gynecology training in Turkey has significantly been affected by the COVID-19 pandemic.


Total penile glans amputation following circumcision: A case report of a dramatic complication.
Circumcision is one of the most commonly performed surgical procedures. As traditional ritual circumcisions are still practiced in the community, in Senegal the majority of circumcisions are performed in hospitals. We report the case of a 9-year-old boy who underwent a total amputation of the glans after a circumcision in a pharmacy by an unqualified agent. A meatoplasty was performed and the postoperative course were uneventful.


Exploring Key Factors Influencing Nursing Students' Cognitive Load and Willingness to Serve Older Adults: Cross-sectional Descriptive Correlational Study.
Virtual learning environments (VLEs) use a virtual environment to support learning activities. VLEs are commonly used to overcome the temporal and spatial restrictions of learning activities held in conventional face-to-face classrooms. In VLEs, students can participate in learning activities using the internet, and teachers can provide assistive learning tools during the process. The purpose of this study was to investigate the relationships among nursing students' mental load, cognitive load, and affective learning outcomes in terms of their willingness to serve older adults in an interaction-based educational virtual reality (VR) learning environment. This study used a cross-sectional method. A total of 130 students participated in interaction-based VR learning and completed related questionnaires. Descriptive and inferential statistics and stepwise regression for data analysis were used. The research results revealed that in the dimension of willingness to use VR learning materials, perceived usefulness received the highest score (mean 4.42, SD 0.45). In the dimension of nursing ability, students scored the highest in information management and application ability to care for case patients (mean 4.35, SD 0.54). Correlation analysis revealed that cognitive load during learning and willingness to serve older adults were negatively correlated, whereas willingness to use VR learning materials was positively correlated with nursing ability and willingness to serve older adults. Analyzing the regression coefficients of predictor variables revealed that willingness to use VR learning materials (β=.23; t2=2.89, P=.005) and cognitive load during learning (β=-.35; t2=-.4.30, P<.001) were predictive factors of nursing students' willingness to serve older adults. This study demonstrated that students' willingness to use VR learning materials and their cognitive load during learning affected their willingness to care for older adults. Therefore, the components of mental or cognitive load generate inconsistent predictive effects on affective variables and willingness to serve older adults.


Accumulation of Toxic Elements in Bone and Bone Marrow of Deer Living in Various Ecosystems. A Case Study of Farmed and Wild-Living Deer.
The aim of the study was to determine the concentrations of toxic elements accumulated in the bone marrow and bones (Cervus elaphus). The studies were carried out on two groups of young stags: farmed (n = 6) and wild (n = 9). Their body weights were measured and bone and bone marrow samples were collected. The concentrations of toxic elements were analyzed using the inductively coupled plasma mass spectrometry technique. The mean aluminum content in the bone marrow and bones of the farmed animals was significantly higher than in the wild group (p < 0.05). The mean concentration of arsenic, barium and lead in the bones of the wild red deer was significantly higher than in the bones of the farmed animals (p < 0.05), while the cadmium concentration in the bones of the farmed red deer exceeded the value determined in the wild animals. A significant difference was found between the mean concentrations of aluminum, arsenic, barium, lead, vanadium and nickel in the bone marrow and bones of both red deer groups (p < 0.05). Although the study involved animals living in an uncontaminated area, the concentrations of some heavy metals were higher than values reported from industrial regions.


Targeting hypoxia in solid and haematological malignancies.
Tumour hypoxia is a known and extensively researched phenomenon that occurs in both solid and haematological malignancies. As cancer cells proliferate, demand for oxygen can outstrip supply reducing tumour oxygenation. In solid tumours this is contributed to by disorganized blood vessel development. Tumour hypoxia is associated with resistance to treatment, more aggressive disease behaviour and an increased likelihood of metastatic progression. It can be measured using both invasive and non-invasive methods to varying degrees of accuracy. The presence of hypoxia stimulates a complex cellular network of downstream factors including Hypoxia Inducible Factor 1 (HIF1), C-X-C motif chemokine 4 (CXCR4) and Hypoxia-inducible glycolytic enzyme hexokinase-2 (HK2) amongst many others. They work by affecting different mechanisms including influencing angiogenesis, treatment resistance, immune surveillance and the ability to metastasize all of which contribute to a more aggressive disease pattern. Tumour hypoxia has been correlated with poorer outcomes and worse prognosis in patients. The correlation between hypoxic microenvironments and poor prognosis has led to an interest in trying to therapeutically target this phenomenon. Various methods have been used to target hypoxic microenvironments. Hypoxia-activated prodrugs (HAPs) are drugs that are only activated within hypoxic environments and these agents have been subject to investigation in several clinical trials. Drugs that target downstream factors of hypoxic environments including HIF inhibitors, mammalian target of rapamycin (mTOR) inhibitors and vascular endothelial growth factor (anti-VEGF) therapies are also in development and being used in combination in clinical trials. Despite promising pre-clinical data, clinical trials of hypoxia targeting strategies have proven challenging. Further understanding of the effect of hypoxia and related molecular mechanisms in human rather than animal models is required to guide novel therapeutic strategies and future trial design. This review will discuss the currently available methods of hypoxia targeting and assessments that may be considered in planning future clinical trials. It will also outline key trials to date in both the solid and haemato-oncology treatment spheres and discuss the limitations that may have impacted on clinical success to date.


Comparison between the effect of plasma and chemical treatments on fiber post surface.
The aim of the present study was to evaluate the fiber post surface after plasma and usual treatments and the adhesion between treated fiber posts and Rely X Unicem resin cement (3M ESPE, St Paul, MN). Flat fiber posts were divided into 6 groups according to surface treatment: silane, hydrofluoric acid, hydrofluoric acid plus silane, plasma polymerization with argon, ethylenediamine plasma (EDA), and the control group. A goniometer was used to measure the contact angle between the groups with water or resin cement. Scanning electron microscopy and electron dispersive spectroscopy were used to examine the topography and chemical changes in the post surfaces after treatment. Push-out tests were performed using a universal testing machine to evaluate the adhesion strength between treated fiber posts and resin cement. In the contact angle with water, the most hydrophilic surface was observed in samples treated with argon plasma, followed by treatments with silane and hydrofluoric acid plus silane. The hydrophobic characteristic was observed with EDA and hydrofluoric acid. The contact angle with dual resin cement showed lower values with argon and EDA, followed by silane and hydrofluoric acid plus silane. Electron dispersive spectroscopy analyses showed chemical modifications in the surface after different treatments although topographic changes were verified only with EDA plasma compared with the control. Push-out results did not show differences between groups compared with the control, except for EDA plasma treatment. Plasma treatment favored the wettability of the post surface by modifying it chemically. Adhesion improvement was only observed after EDA treatment.


Is the ALS a motor neuron disease or a hematopoietic stem cell disease?
Amyotrophic lateral sclerosis (ALS) is also known as motor neuron disease (MND) or Lou Gehrig's disease. It is a fatal neurodegenerative disease the cause of which is not clear. The effective therapy is absent. ALS is diagnosed through clinical examination and neurophysiologic tests. Clinically, the symptoms manifest when about 80% of motor neurons are dead. The hematopoietic stem cells are isolated through administration of the granulocyte colony-stimulating factor from three groups: group 1 of 62 ALS cases, group 2 of 54 ALS-free healthy donors and group 3 of 6 ALS-free ALS-family members. The expression of HLA-DR, CD38, CD117, CD13, CD33, CD56, Thy-1, CD45, СD10, CD71 was assessed in 86 samples of HSCs in ALS group, 61 samples of HSCs in healthy group and 6 samples from ALS-free ALS-family members by the multiparameter flow cytometry. The obtained immunophenotypic profiles of HSCs membrane antigens of the ALS group significantly differ from the ALS-free group, while the immunophenotypic profiles of HSCs membrane antigens of the ALS-family members group are close to the ALS group. We suppose that the ALS onset as the disease of HSCs and manifests in the genome and proteome of the HSCs. Such immunophenotypic profiling might permit identification of ALS-specific immune insufficiency and become a tool for early diagnostics of the ALS before clinical manifestation of the disease. New options of the updated therapy of ALS might be developed or corrected considering this new evidence. Further research with larger samples and deeper examination is required.


Microcystin-LR degradation kinetics during chlorination: Role of water quality conditions.
Microcystin-LR (MCLR) produced during certain cyanobacteria blooms can contaminate drinking water sources and pose a threat to public health. Previous studies of MCLR degradation by free chlorine may have artifacts from using strong reducing agents to quench chlorination reactions, and they also have not explored the influence of water quality characteristics such as pH, alkalinity, temperature and dissolved organic matter (DOM). Using a novel quencher, 1,3,5-trimethoxybenzene (TMB), the apparent MCLR degradation rate constants were found to be higher than those obtained with thiosulfate (S2O32-), a traditionally used strong reducing quencher. Thiosulfate converted N-chlorinated MCLR degradation products back to the parent MCLR, thereby underestimating MCLR loss over time. The second-order rate constants for HOCl (kHOCl) and OCl- (kOCl-) during chlorination of MCLR were determined to be 72 ± 13 and 28 ± 1.8 M-1s-1, respectively, allowing for determination of the apparent MCLR rate constants (kapp,MCLR) for any known pH condition. The MCLR reaction with free chlorine was strongly affected by temperature and the presence of DOM, while changes in ionic strength and alkalinity had little effect. Free chlorine in the presence of DOM, originating from both terrestrial and microbial sources, exhibited two-stage decay. The initial chlorine demand in the first 15 s of reaction can be determined by the dissolved organic carbon (DOC) concentration (initial chlorine demand = 1.8 × DOC), and the second-order rate constants for the later slower decay correlated well with SUVA254 (kapp,DOM = 0.73 × SUVA254 - 0.41). The results yielded a practical model to predict the decay of MCLR during chlorination of waters with varied water quality characteristics.


Designed multi-domain protein as a carrier of nucleic acids into cells.
Protein-based nucleic acid carriers offer attractive possibilities to enhance in vitro and in vivo gene delivery to combat diseases. A multi-domain fusion protein, namely TAT-NLS-Mu, designated as TNM, has been designed, cloned, heterologously expressed in E. coli and purified to homogeneity by affinity chromatography. The recombinant chimera TNM harbors three epitopes, a cell-penetrating (TAT) domain, a nuclear localization domain comprising of three nuclear localization sequence (NLS) motifs in tandem and a DNA-binding (Mu) domain. Complexes prepared by combining plasmid DNA with TNM (DP) transfect MCF-7, COS, CHO and HepG2 cells. Ternary complexes prepared with DNA, protein and cationic lipid (DPL) resulted in ~5-7 fold enhancement in reporter gene expression over the DP alone. Treatment of cells with chloroquine during transfection, with DP complexes, resulted in remarkable increases in reporter gene expression suggesting the involvement of endosomal compartments in the uptake process. Interestingly, DPL prepared with Lipofectin or 1, 2-Dioleoyl-3-Trimethylammonium-Propane (DOTAP) exhibited enhanced transfection in the presence of serum in MCF-7 and HepG2 cells. Microinjection of DP complexes, with and without NLS sequence, into the cytoplasm and nucleus of smooth muscle cells (SMC) indicated that the presence of NLS sequence in protein carrier significantly enhanced transgene expression. Together the data suggest that modular design of proteins is a promising method to develop gene delivery carriers and also the role of NLS epitopes in mediating nuclear transfer of DNA complexes into various cell types.


[Male infertility].
In recent years approximately 8% of the newborn Danes have been conceived by infertility treatment, and approximately half of the cases are due to male subfertility. Male infertility can be caused by several factors, and only in about half of the cases is it possible to disclose an aetiological explanation. It is important to elucidate possible reasons for male infertility as low semen quality might be a symptom of pituitary dysfunction, genetic disorders or testicular cancer.


Expanded progeny cells of diffuse cytotoxic bone marrow-derived colonies.
Bone marrow-derived (colony-stimulating factor [CSF]-dependent) diffuse colonies have been shown to include colonies with cytotoxic activity. Such diffuse colonies were expanded for 6-8 weeks in liquid culture medium in the presence of pokeweed mitogen- or concanavalin A-conditioned spleen cell medium (CM). The morphology of the expanded diffuse colony cells (EDCC) was like that of early myelocytic cells. EDCC lost their cytotoxic capacity when expanded, but the cytotoxicity could be reinduced by pretreatment of the colonies with interferon or phorbol ester. Traditional sources of mouse or human CSF such as lung CM, placenta CM and human mononuclear cell CM did not support proliferation of EDCC, whereas partly purified interleukin-3 (IL-3), lacking CSF and IL-2, was stimulatory for EDCC. Thus, the stimulatory factor for EDCC was not CSF but a factor closely related to IL-3. Monoclonal antibodies against T lymphocytes or macrophages did not bind to EDCC. EDCC did not have Fc receptors, but 10% of the cells were positive for a monoclonal anti-Ia antibody. All EDCC were positive for alpha NAE and NASDCI esterases but negative for acid and alkaline phosphatases and peroxidase reactivity; less than 2% of the cells showed metachromatic staining with toluidine. Ultrastructurally, EDCC showed various degrees of cell differentiation but absence of specific cytoplasmatic characteristics such as neutrophilic, eosinophilic, basophilic and mast cell granules. Current work aims to define factors and conditions necessary for the induction of differentiation in these immature monomyelocytic cells.


Establishment of an efficient transformation system and its application in regulatory mechanism analysis of biological macromolecules in tea plants.
Tea (Camellia sinensis), one of the most important beverage crops originated from China and is now cultivated worldwide, provides numerous secondary metabolites that account for its health benefits and rich flavor. However, the lack of an efficient and reliable genetic transformation system has seriously hindered the gene function investigation and precise breeding of C. sinensis. In this study, we established a highly efficient, labor-saving, and cost-effective Agrobacterium rhizogenes-mediated hairy roots genetic transformation system for C. sinensis, which can be used for gene overexpression and genome editing. The established transformation system was simple to operate, bypassing tissue culture and antibiotic screening, and only took two months to complete. We used this system to conduct function analysis of transcription factor CsMYB73 and found that CsMYB73 negatively regulates L-theanine synthesis in tea plant. Additionally, callus formation was successfully induced using transgenic roots, and the transgenic callus exhibited normal chlorophyll production, enabling the study of the corresponding biological functions. Furthermore, this genetic transformation system was effective for multiple C. sinensis varieties and other woody plant species. By overcoming technical obstacles such as low efficiency, long experimental periods, and high costs, this genetic transformation will be a valuable tool for routine gene investigation and precise breeding in tea plants.


Autotrophic synthesis of polyhydroxyalkanoates by the bacteria Ralstonia eutropha in the presence of carbon monoxide.
It has been found that the carbon monoxide (CO)-resistant strain of the hydrogen bacteria Ralstonia eutropha B5786 is able to synthesise polyhydroxyalkanoates (PHAs) in the presence of CO under autotrophic conditions. This strain, grown on model gas mixtures containing 5-25% CO (v/v), accumulates up to 70-75% (of absolutely dry matter) PHA, without significant variation in the yield coefficient on hydrogen. No suppression of the activities of the key enzymes of PHA synthesis ( beta-ketothiolase, acetoacetyl-CoA-reductase, butyrate dehydrogenase and poly-3-hydroxybutyrate synthase) was recorded. The PHA synthesised is a co-polymer containing mostly beta-hydroxybutyrate (more than 99 mol%) with trace amounts of beta-hydroxyvalerate. The investigated properties of the polymer (molecular weight, crystallinity, temperature characteristics) do not differ from those of the polymer synthesised on electrolytic hydrogen.