Profiling of membrane protein variants in a baculovirus system by coupling cell-surface detection with small-scale parallel expression.
Production of structure-grade mammalian membrane proteins in substantial quantities has been hindered by a lack of methods for effectively profiling multiple constructs expression in higher eukaryotic systems such as insect or mammalian cells. To address this problem, a specialized small-scale eukaryotic expression platform by Thomson Instrument Company (Vertiga-IM) was developed and used in tandem with a Guava EasyCyte microcapillary 96-well cytometer to monitor cell density and health and evaluate membrane protein expression. Two proof of concept experiments were conducted using the human beta(2)-adrenergic receptor (beta(2)AR) and the gap junction protein connexin26 (Cx26) in a baculovirus expression system. First, cell surface expression was used to assess the expression levels of 14 beta(2)AR truncation variants expressed using the Vertiga-IM shaker. Three of these variants were then compared to wild-type beta(2)AR using three metrics: cell surface expression, saturation ligand binding and protein immunoblot analysis of dodecylmaltoside extracted material. Second, a series of systematic Cx26 truncation variants were evaluated for expression by protein immunoblot analysis. The cumulative results for these two systems show that the Vertiga-IM instrument can be used effectively in the parallel insect cell microexpression of membrane protein variants, and that the expression of cell surface molecules as monitored with the Guava EasyCyte instrument can be used to rapidly assess the production of properly folded proteins in the baculovirus expression system. This approach expedites the in vitro evaluation of a large number of mammalian membrane protein variants.


Colonic hemorrhage from a solitary minute ulcer. Report of three cases.
Three patients with massive lower gastrointestinal bleeding are reported. In all cases, the bleeding source was localized by emergency selective mesenteric angiography. The histologic lesion found in the resected specimen consisted of a minute mucosal ulcer with an abnormally large eroded submucosal artery without evidence of true angiodysplastic changes. The clinicopathological picture is similar to the rare solitary stomach ulceration, described as "Exulceratio simplex Dieulafoy."


A case of Creutzfeldt-Jakob disease related to familial retinitis pigmentosa patients.
Similarities between serological alterations and retinal degeneration occurring in natural and experimental CJD and in some forms of human retinal degeneration have recently been reported. In the present paper a family from an areal focal accumulation of CJD in Central Slovakia with 1 histopathologically verified case of Creutzfeldt-Jakob disease and 2 case of retinitis pigmentosa is described. Neuropathological and epidemiological data obtained in investigated patients are discussed from the point of view of a possible relationship between the slow virus infections caused by unconventional agents and degenerative disease affecting the ocular system.


Effects of internal limiting membrane peeling in vitrectomy on diabetic cystoid macular edema patients.
We evaluated the effects of the peeling of the internal limiting membrane (ILM) during vitrectomy in diabetic cystoid macular edema (CME) patients. Visual outcome and intraoperative and postoperative complications were evaluated retrospectively in 84 CME patients (100 eyes), all of whom had been followed for at least 1 year postoperatively. Before January 2001, we did not perform ILM peeling at our hospitals; 57 patients (66 eyes) treated before 2001 were included in this retrospective study as the non-peeling group. After January 2001, ILM peeling was performed in 27 (34 eyes) CME patients, who were included in this study as the peeling group. In the peeling group, indocyanine green (ICG) staining was performed at the time of ILM peeling. Visual acuity improved significantly after vitrectomy regardless of ILM peeling. Visual acuity improved gradually from 6 months to 1 year after the operation, and improved further at the final observation point in both groups. Visual acuity did not differ significantly between the two groups at any time point. There was no difference in the incidence of intraoperative and postoperative complications between the two groups. There were no adverse events associated with ICG-assisted ILM peeling. Visual acuity improved with vitrectomy for diabetic cystoid macular edema in both groups. ILM peeling was not found to improve visual acuity postoperatively.


Pediatric scapular fractures and associated injuries following blunt chest trauma.
Scapular fractures in the pediatric population are rare, and medical literature is lacking regarding these specific injuries in the pediatric population. Prior studies have shown that scapular fractures resulting from blunt chest trauma have been associated with significant morbidities in adults, and that a majority of scapular fractures are missed on chest X-ray (CXR) and seen on computerized tomography only (SOCTO). Further guidance is needed regarding the prevalence of coinciding injuries in the pediatric population and the modality for diagnosis. The primary objectives of this study were to assess 1) the frequency of scapular fractures following blunt trauma in the pediatric cohort, 2) the frequency of other associated thoracic injuries, 3) the proportion on scapular fractures SOCTO. We conducted a retrospective cohort study with data obtained from our study site's Trauma Registry. Patients under 18 years receiving both a CXR and chest CT following blunt trauma or any patient diagnosed with a scapular fracture by any modality from January 2009 to December 2019 were included. Primary outcome variables were the presence of a scapular fracture diagnosed by any modality, absence of scapular fracture, and scapular fractures SOCTO. Charts were also reviewed for the following concurring injuries: 1) contusion/atelectasis, 2) pneumothorax, 3) hemothorax, 4) rib fracture, 5) other fracture, 6) vascular injury, 7) mediastinal injury, 8) diaphragm rupture, 9) foreign body, 10) incidental finding. Of 12,826 charts of pediatric patients with blunt chest trauma, 1405 obtained both CXR and chest CT. Sixty (0.47%) were diagnosed with scapular fracture, and 48 (73.3%) of the fractures were SOCTO. The most commonly associated injuries were other fracture (88.3%), lung contusion/atelectasis (78.3%), pneumothorax (58.3%) and rib fracture (58.3%). Patients with scapular fractures had higher injury severity scores (ISS) and more frequently required surgery for other intrathoracic injuries. Only five patients required surgical management of the scapular fracture with the rest managed conservatively. Pediatric scapular fractures are rare and are often associated with other intrathoracic injury. A majority of scapula fractures are missed on CXR, but identification of the injury did not change management as most were treated conservatively.


Effect of exercise on upper extremity pain and dysfunction in head and neck cancer survivors: a randomized controlled trial.
Shoulder pain and disability are well recognized complications associated with surgery for head and neck cancer. This study was designed to examine the effects of progressive resistance exercise training (PRET) on upper extremity pain and dysfunction in postsurgical head and neck cancer survivors. Fifty-two head and neck cancer survivors were assigned randomly to PRET (n = 27) or a standardized therapeutic exercise protocol (TP) (n = 25) for 12 weeks. The primary endpoint was change in patient-rated shoulder pain and disability from baseline to postintervention. Secondary endpoints were upper extremity strength and endurance, range of motion, fatigue, and quality of life. Follow-up assessment for the primary outcome was 92%, and adherence to the supervised PRET and TP programs were 95% and 87%, respectively. On the basis of intention-to-treat analyses, PRET was superior to TP for improving shoulder pain and disability (-9.6; 95% confidence interval [95% CI], -16.4 to -4.5; P = .001), upper extremity strength (+10.8 kg; 95% CI, 5.4-16.2 kg; P < .001), and upper extremity endurance (+194 repetitions x kg; 95% CI, 10-378 repetitions x kg; P = .039). Changes in neck dissection impairment, fatigue, and quality of life favored the PRET group but did not reach statistical significance. The PRET program significantly reduced shoulder pain and disability and improved upper extremity muscular strength and endurance in head and neck cancer survivors who had shoulder dysfunction because of spinal accessory nerve damage. Clinicians should consider the addition of PRET in the rehabilitation of postsurgical head and neck cancer survivors.


Structure-function relationship of soleus muscle fibres from the rhesus monkey.
Functional and structural properties of rhesus monkey skinned fibres were studied in order to examine the relationship between calcium/strontium (Ca/Sr) activation characteristics and protein composition. The fibres were classified according to their Ca/Sr affinity into slow (61%) and fast groups (39%). According to the myosin isoform composition, two additional hybrid types were defined. Thus, four profiles were characterized: two corresponding to slow (S) and fast (F) isoforms and two corresponding to a mixed proportion of slow and fast isoforms. They were called hybrid slow (HS) or hybrid fast (HF) based on the predominant myosin isoform. Tension/pCa parameters and maximal shortening velocities were determined. S fibres showed a higher pCa threshold and affinity as well as shallower slopes of their tension/pCa curve than did F fibres. HS and HF fibres exhibited tension/pCa curves which were positioned close to those of S and F fibres, respectively. No significant difference was observed between S and HS fibres or between F and HF fibres. Maximal shortening velocity values were higher for fibres expressing predominantly fast myosin isoforms. We suggest than when both S and F isoforms of myofibrillar proteins are expressed in a muscle fibre, the functional properties are mainly governed by the predominant isoform.


Pain and desire for hastened death in terminally ill cancer patients.
The purpose of this study was to assess the relationship between pain and the desire for hastened death in terminally ill cancer patients. The participants were 120 terminally ill cancer patients under palliative treatment from June 2003 to November 2004. Patients completed a pain assessment tool, the Greek Brief Pain Inventory (G-BPI), and a self-report measure of the desire for hastened death, the Greek Schedule of Attitudes Toward Hastened Death (G-SAHD). Moderate but statistically significant associations were found between some of the severity and interference items of G-BPI and G-SAHD; more specifically, between G-SAHD and G-BPI3, "worst pain in the last 24 hours" (r = 0.279, P = .002); G-SAHD and G-BPI4, "least pain in the last 24 hours" (r = 0.253, P = .005); and G-SAHD and G-BPI5, "average pain in the last 24 hours" (r = 0.283, P = .002). A stronger association was revealed between G-SAHD and G-BPI8, "relief provided by pain treatment and medications in the last 24 hours" (r = -0.326, P = .000). Multiple regression analyses including the enter model and the forward model were conducted. According to the enter model, the strongest predictors of hastened death were items G-BPI6, "current pain"; G-BPI8, "relief provided by pain treatment and medications in the last 24 hours"; G-BPI9i, "interference of pain in general activity"; and G-BPI9iii, "interference of pain in walking." According to the forward model, significant predictors of the desire for death were items G-BPI5, "average pain in the last 24 hours"; G-BPI6, "current pain"; G-BPI9i, "interference of pain in general activity"; and G-BPI9ii, "interference of pain in mood," all of which were statistically significant (P = .000-.042). Pain appeared to have a statistically significant relationship with the desire for hastened death. Effective treatment by healthcare professionals should be provided to reduce pain and cancer-related symptoms as well as the desire for hastened death.


Purification of mitochondrial NADH dehydrogenase from Drosophila hydei and comparison with the 'heat-shock' polypeptides.
Mitochondrial NADH dehydrogenase (NADH:(acceptor) oxidoreductase, EC .6.99.3) from either Drosophila hydei larvae or embryos has been purified 150- and 120-fold, respectively. The purified enzyme appeared homogeneous and showed a molecular weight of 57 000. The molecular weight of the nondenatured enzyme was 79 000. On isoelectro-focussing of the preparation, two fractions were observed, a major one with an isoelectric point of 6.2 and a minor fraction with an isoelectric point of 4.9. Straight-line kinetics in Lineweaver-Burk plots were observed for the purified enzyme with a Km of 0.040 mM. The Km was not changed during the purification procedure, suggesting that the enzyme was not denatured or inactivated. The pH optimum of the purified enzyme was 5.6. The molecular weight of the purified mitochondrial NADH dehydrogenase does not correspond to that of one of the 'heat-shock' polypeptides.


Transdermal drug transport and metabolism. I. Comparison of in vitro and in vivo results.
Using excised human skin and tissue grafted to athymic mice, the in vitro and in vivo delivery and metabolism of a salicylate diester were compared. Concentration profiles of this drug and its metabolites were obtained for the outer several hundred microns of the skin. These results show significant differences in the extent of enzymatic cleavage and distribution of metabolites between in vitro and in vivo studies. Furthermore, these data suggest that in vitro results may overestimate metabolism because of increased enzymatic activity and/or decreased capillary removal.


In Situ Antegrade Laser Fenestrations During Endovascular Aortic Repair.
The technique and preliminary outcomes are reported for laser fenestrated endografting (LfEVAR), developed as an alternative procedure for endovascular repair of thoraco-abdominal aortic aneurysms (TAAAs), type IA endoleaks (T1AELs), and pararenal aortic aneurysms (PAAAs). Patients with TAAA, T1AEL, and PAAA considered unfit for open repair by a multidisciplinary team and who could not benefit from a custom made device were selected. LfEVAR is a physician modified technique requiring sequential steps. After preliminary stenting of each target artery, a standard stent graft was deployed in the aorta. Laser fenestration was performed using a steerable sheath positioned in the stent graft lumen in front of each target artery ostium. After progressive dilation of the laser fenestration, bridging stents were placed and flared to insure accurate perfusion of the visceral arteries. Between August 2015 and March 2017, 16 consecutive patients were treated using LfEVAR including two TAAAs, four T1AELs, and 10 PAAAs. Thirty-three laser fenestrations were performed with a 94.3% success rate for visceral artery fenestration. The median ischaemic was 7.5 minutes for the superior mesenteric artery, 48 and 50 minutes for the left and right renal arteries, and 125 minutes for the coeliac trunk. Four secondary procedures were required: two endoleaks (type II and III), and two related to fenestrations: one renal stent dislocation, and one renal artery stent stenosis. During a mean follow up of 17 months, no deaths occurred. Follow up computed tomography (CT) scans performed at 30 days, 6, 12, and 18 months were satisfactory (4 endoleaks and two aneurysm enlargements were observed). The follow up CT scan imaging demonstrated 97% primary patency. In situ antegrade LfEVAR is feasible and a safe alternative option for high risk inoperable patients or for those not eligible for custom made device endografting, particularly in emergency cases.


Morphological and molecular characterization of Neotropic Lymnaeidae (Gastropoda: Lymnaeoidea), vectors of fasciolosis.
Lymnaeidae play a crucial role in the transmission of fasciolosis, a disease of medical and veterinary importance. In the Neotropic, a region where fasciolosis is emergent, eight Lymnaeidae species are currently considered valid. However, our knowledge of the diversity of this taxon is hindered by the fact that lymnaeids exhibit extremely homogeneous anatomical traits. Because most species are difficult to identify using classic taxonomy, it is difficult to establish an epidemiological risk map of fasciolosis in the Neotropic. In this paper, we contribute to our understanding of the diversity of lymnaeids in this region of the world. We perform conchological, anatomical and DNA-based analyses (phylogeny and barcoding) of almost all species of Lymnaeidae inhabiting the Neotropic to compare the reliability of classic taxonomy and DNA-based approaches, and to delimitate species boundaries. Our results demonstrate that while morphological traits are unable to separate phenotypically similar species, DNA-based approaches unambiguously ascribe individuals to one species or another. We demonstrate that a taxon found in Colombia and Venezuela (Galba sp.) is closely related yet sufficiently divergent from Galba truncatula, G. humilis, G. cousini, G. cubensis, G. neotropica and G. viatrix to be considered as a different species. In addition, barcode results suggest that G. cubensis, G. neotropica and G. viatrix might be conspecifics. We conclude that conchological and anatomical characters are uninformative to identify closely related species of Lymnaeidae and that DNA-based approaches should be preferred.


Determination of BAY x 7195, a novel leukotriene D4 antagonist, in human plasma by high-performance liquid chromatography with post-column photo derivatisation and fluorescence detection.
Methods to determine plasma concentrations of the leukotriene D4 antagonist BAY x 7195 by HPLC with post-column photo derivatisation and fluorescence detection are described. Following dilution and centrifugation plasma supernatant is injected onto the HPLC system allowing the selective determination of the drug with a limit of quantitation (LOQ) of 10 micrograms/l (method A). Sensitivity was further enhanced to a LOQ of 0.6 microgram/l by employing solid-phase extraction whereby the analyte concentration in the injection solution was increased (method B). Data on recovery, accuracy and precision of both methods throughout the working range are presented. BAY x 7195 is stable in plasma after repeated freeze-thaw cycles and upon storage at -20 degrees C for at least 13 months. Method A was applied to a clinical study with oral administration of 250 mg BAY x 7195 where ca. 1% of the maximum plasma concentrations still could be accurately and precisely quantified. Method B was employed to determine the drug in plasma after administration of 1 mg as aerosol.


Phalloidin poisoning of isolated hepatocytes: inhibition of protein synthesis.
Phalloidin added to isolated hepatocytes inhibits amino acid incorporation into protein. Inhibition is very strong with 20 micrograms/ml phalloidin and occurs after a lag period of about 30 min. After this period, poisoned hepatocytes also show morphological changes in their plasma membrane.


Clinical management and impact of scarlet fever in the modern era: findings from a cross-sectional study of cases in London, 2018-2019.
In response to increasing incidence of scarlet fever and wider outbreaks of group A streptococcal infections in London, we aimed to characterise the epidemiology, symptoms, management and consequences of scarlet fever, and to identify factors associated with delayed diagnosis. Cross-sectional community-based study of children with scarlet fever notified to London's three Health Protection Teams, 2018-2019. From 2575 directly invited notified cases plus invitations via parental networks at 410 schools/nurseries with notified outbreaks of confirmed/probable scarlet fever, we received 477 responses (19% of those directly invited), of which 412 met the case definition. Median age was 4 years (range <1 to 16), 48% were female, and 70% were of white ethnicity. Preplanned measures included quantitative description of case demographics, symptoms, care-seeking, and clinical, social, and economic impact on cases and households. After survey completion, secondary analyses of factors associated with delayed diagnosis (by logistic regression) and consequences of delayed diagnosis (by Cox's regression), and qualitative analysis of free text comments were added. Rash was reported for 89% of cases, but followed onset of other symptoms for 71%, with a median 1-day delay. Pattern of onset varied with age: sore throat was more common at onset among children 5 years and older (OR3.1, 95% CI 1.9 to 5.0). At first consultation, for 28%, scarlet fever was not considered: in these cases, symptoms were frequently attributed to viral infection (60%, 64/106). Delay in diagnosis beyond first consultation occurred more frequently among children aged 5+ who presented with sore throat (OR 2.8 vs 5+without sore throat; 95% CI 1.3 to 5.8). Cases with delayed diagnosis took, on average, 1 day longer to return to baseline activities. Scarlet fever may be initially overlooked, especially among older children presenting with sore throat. Raising awareness among carers and practitioners may aid identification and timely treatment.


Treatment of chronic urticaria with a beta2-adrenergic stimulant.
Twenty-four patients with chronic utricaria resistant to conventional therapy were treated orally with terbutaline, a specific beta2 stimulant. This therapy was much more effective than antihistamines. At the dose of 1-25 mg three times a day, there were few side effects. The efficacy was independent of the IgE levels; it seemed greater in idiopathic cases than in cholinergic or pressure induced urticaria.


Conformational changes of lysozyme refolding intermediates and implications for aggregation and renaturation.
It is believed that denatured-reduced lysozyme rapidly forms aggregates during refolding process, which is often worked around by operating at low protein concentrations or in the presence of aggregation inhibitors. However, we found that low concentration buffer alone could efficiently suppress aggregation. Based on this finding, stable equilibrium intermediate states of denatured-reduced lysozyme containing eight free SH groups were obtained in the absence of redox reagents in buffer of low concentrations alone at neutral or mildly alkaline pH. Transition in the secondary structure of the intermediate from native-like to beta-sheet was observed by circular dichroism (CD) as conditions were varied. Dynamic light scattering and ANS-binding studies showed that the self-association accompanied the conformational change and the structure rich in beta-sheet was the intermediate state for aggregation, which could form either amyloid protofibril or amorphous aggregates under different conditions as detected by Electron Microscopy. Combining the results obtained from activity analysis, RP-HPLC and CD, we show that the activity recovery was closely related to the conformation of the refolding intermediate, and buffer of very low concentration (e.g. 10mM) alone could efficiently promote correct refolding by maintaining the native-like secondary structure of the intermediate state. This study reveals reasons for lysozyme aggregation and puts new insights into protein and inclusion body refolding.


Highly Efficient Organic Hole Transporting Materials for Perovskite and Organic Solar Cells with Long-Term Stability.
Small molecules based on N-atom-linked phenylcarbazole-fluorene as the main scaffold, end-capped with spirobifluorene derivatives, are developed as organic hole-transporting materials for highly efficient perovskite solar cells (PSCs) and bulk heterojunction (BHJ) inverted organic solar cells (IOSCs). The CzPAF-SBF-based devices show remarkable device performance with excellent long-term stability in PSCs and BHJ IOSCs with a maximum PCE of 17.21% and 7.93%, respectively.


Subvalvular aortic stenosis diagnosed by 3D transesophageal echocardiography.
The patient was a 13-year-old male with chief complaints of exertional chest pain and dyspnea. Cardiac murmur was suspected in a medical checkup at 1 month old, at which time he was diagnosed with subvalvular aortic stenosis. He had subsequently been under follow-up observation at a nearby hospital for subvalvular aortic stenosis. He was admitted to our department for surgery due to aggravation of symptoms that had occurred over the previous year. Transthoracic echocardiography after admission showed an abnormal structure in the subvalvular aortic area, and the maximum pressure gradient between the left ventricle and aortic valve was 84 mmHg. The preoperative valve area was 0.71 cm(2), as measured by the Doppler method. Measurement of valve area by the trace method was difficult. Transesophageal echocardiography (TEE) showed a septum-like structure extending from the ventricular septum in the subvalvular area. On 3D TEE, the valve areas in the systolic and diastolic phases were 0.86 and 0.49 cm(2), respectively. Postoperative echocardiography showed resection of the structure in the subvalvular area, and the postoperative course was favorable.


Asthenopia as the presenting symptom in advance allergic fungal sinusitis.
A 14-year-old male presented to the ophthalmology clinic with a history of asthenopia. Laboratory, radiological and histopathological studies confirmed the diagnosis of allergic fungal sinusitis resulting in lateral displacement of the medial rectus muscles. Symptoms improved and near point of convergence recovered after surgical endoscopic decompression of the ethmoidal and maxillary sinuses.


Age-related changes in the morphology and immunophenotype of spontaneous lymphomas of SJL/N mice.
Forty-four SJL mice were killed at intervals to follow the spontaneous development of lymphomas. These were detected in 28 of 38 mice aged 24 weeks or more. Immunoperoxidase stains were applied on cryostat sections of lymphomas from various sites (spleen, mesenteric lymph nodes, Peyer's patches, and other lymph nodes), sampled from 19 mice of different age groups, to study tumour immunophenotype in correlation with histological features. With a panel of antibodies against Thy 1.2, Lyt 1, and Lyt 2, and the mouse immunoglobulin heavy and light chains, all the 19 tumours were shown to be B-cell derived. This was further supported by the detection of Ia antigens on the tumour cells. Six mice had plasmacytoid or predominantly plasmacytoid tumours expressing gamma kappa. Three mice had mixed plasmacytoid and pleomorphic tumours which also showed gamma kappa positivity. The ages of these nine mice ranged from 24 to 44 weeks with a median of 36 weeks. Pleomorphic tumours were found in ten mice (age range = 36-52 weeks, median = 44 weeks). These tumours were more heterogeneous, expressing either alpha, mu, or gamma, or with loss of heavy chain expression altogether. These age-related changes are probably a sign of progressive loss of differentiation in B lymphomas.


Current thinking in lower third molar surgery.
The removal of lower third molar teeth is one of the most common surgical procedures performed worldwide, but many concepts in this surgery have been unclear and have engendered different opinions. This paper aims to review current thinking in certain pertinent aspects of this surgery to update the reader on the most current research and synthesise it to make clinical recommendations. Topics covered include preoperative imaging, timing of removal, flap design, lingual retraction, coronectomy, lingual split, closure techniques, and use of antibiotics, steroids, and drains.


Detrended fluctuation analysis as a statistical method to study ion single channel signal.
The scaling behaviors of ion single channel current signal time series could be analyzed by means of detrended fluctuation analysis. Nevertheless, the statistical analysis of an ionic current signal recorded from voltage dependence K+ single channel is presented. The detrended fluctuation analysis (DFA) exponent alpha is significantly larger than 0.5, as (DFA) exponents were calculated for 4 different pipette potentials in rat dorsal root ganglion neurons. alpha=0.9475+/-0.006 for V=-30 mV; alpha=0.958+/-0.004 for V=-40 mV; alpha=0.966+/-0.005 for V=-50 mV; alpha=0.971+/-0.03 for V=-60 mV. The scaling exponents for different pipette potentials reveal the existence of memory in ion channels, and at the same time, the memory in ion channel depends on the pipette potential. The result of Markovian model data showed that it had different DFA exponent alpha which indicates that long-range correlation effect is present amongst the continued conducting states of the ion channel. Thus a scaling exponent description is found to characterize the fluctuation properties of the non-linear behaviors of ion channel kinetics regardless of whether the channel is in open or closed state.


A two-step purification of mouse liver glyoxalase I and evidence of its dimeric constitution.
Glyoxalase I (S-lactoyl-glutathione methylglyoxal-lyase (isomerizing), EC 126.96.36.199) was purified from Swiss mouse liver to homogeneity by a rapid, two-step procedure involving hydrophobic and affinity chromatography. Homogeneity was established by multiple electrophoretic determinations and by sedimentation equilibrium centrifugation. The purified enzyme exhibited a specific activity of 944 I.U./mg protein an has a molecular weight of 43 000. The enzyme was shown to be a dimer by sodium dodecyl sulfate disc gel electrophoresis and is apparently composed of identical subunits of molecular weights approximating 21 500.


Survival comparison between superficialization of the brachial artery and tunneled central venous catheter placement in hemodialysis patients with heart failure: A retrospective study.
It remains unclear which vascular access provides better survival in hemodialysis patients with heart failure, superficialization of the brachial artery (SBA), or tunneled central venous catheter (TCVC). We retrospectively followed up 60 hemodialysis patients with heart failure who underwent SBA (n = 36) or TCVC placement (n = 24). During the median 2.2-year follow-up period, 36 patients died. The median survival time was significantly longer for the SBA group than for the TCVC group (5.7 vs 1.7 years; P < .05, log-rank test). A multivariate-adjusted Cox regression analysis showed that SBA was associated with a reduced risk of all-cause death (hazard ratio [HR] 0.30; 95% confidence interval [CI] 0.14-0.65). In the cohort of propensity score-matched 15 pairs, patients with SBA experienced fewer all-cause deaths (HR 0.29; 95% CI 0.10-0.77). Our study suggests that SBA is an alternative option in hemodialysis patients with heart failure.


HPLC quantification of two isomeric triterpenic acids isolated from Mitracarpus scaber and antimicrobial activity on Dermatophilus congolensis.
Oleanolic (OA) and ursolic acids (UA) were isolated for the first time from the alcoholic extract of Mitracarpus scaber possessing antimicrobial effects on Dermatophilus congolensis. These two triterpenic acids were also active (MIC 15 microg/ml) on this causative agent of dermatophilosis in African animals. To quantify OA and UA in M. scaber, a new, simple and rapid high-performance liquid chromatography (HPLC) method compatible with MS detection was developed and validated. The mobile phase acetonitrile:H2O (85:15, v/v) was pumped through a C18 octadecylsilyl silica column at a flow rate of 0.6 ml/min and the eluate was monitored at 215 nm. The calibration curves constructed between 0.5 and 10 microg/ml showed linear relationships with good R2 values. The developed method was precise and reproducible with relative standard deviations (RSD) for these two active constituents between 0.22-2.06% (intraday) and 1.61-3.72% (interday) for concentrations from 0.5 to 6 microg/ml. Limits of detection and quantification were, respectively, 0.2 and 0.5 microg/ml.


Evaluation of flexor pollicis longus tendon attrition using color Doppler imaging after volar plate fixation for distal radius fracture.
Early diagnosis of attritional wear of the flexor pollicis longus (FPL) tendon is essential in preventing subsequent tendon rupture. There are currently few objective methods of assessing FPL attrition. We hypothesized that color Doppler imaging could visibly detect FPL tendon attrition, and analyzed our results. We evaluated ultrasound imaging of the contact between the FPL tendon and a volar locking plate using the real-time B-mode and Doppler waveforms of the FPL tendon using the continuous Doppler wave mode in 40 patients who underwent fixation of the distal volar locking plate for distal radius fracture. Twenty out of 40 patients underwent plate removal surgery after ultrasound evaluation. We also assessed the relationship between the Doppler waveforms and attrition of the FPL tendon in these 20 patients. Based on the ultrasound findings (n = 40), we divided Doppler waveforms of the FPL tendon into three categories: type 1, spindle wave; type 2, spindle wave with spike; and type 3, spike wave. There were 23, 11, and six patients with type 1, 2, and 3 waveforms in the affected hand, respectively. There were 37 patients with type 1, three with type 2, and no patient with type 3 waveforms in the contralateral wrist. Of the 20 patients who underwent plate removal, five had type 3 waveforms. We found tendon fraying or partial tears in three of these five patients. In addition, all five patients showed changes to type 1 or 2 waveforms after plate removal. None of the other 15 patients with type 1 or 2 waveforms had any tendon injuries during plate removal. Spike Doppler waveform can indicate abnormal findings, and may be a useful method to predict tendon attrition, because of its visibility. Level III evidence.


[Clinico-endoscopic characteristics and laboratory diagnosis of chronic Campylobacter gastritis in childhood].
Altogether 31 children with chronic gastritis were examined. Three diagnostic tests were employed for detecting Campylobacter pylori. The clinical signs of the disease and the endoscopic features of the gastric mucosa are described.


Validation of the PROMIS physical function measures in a diverse US population-based cohort of cancer patients.
To evaluate the validity of the Patient-Reported Outcomes Measurement Information System (PROMIS) physical function measures in a diverse, population-based cancer sample. Cancer patients 6-13 months post-diagnosis (n = 4840) were recruited for the Measuring Your Health study. Participants were diagnosed between 2010 and 2013 with non-Hodgkin lymphoma or cancers of the colorectum, lung, breast, uterus, cervix, or prostate. Four PROMIS physical function short forms (4a, 6b, 10a, and 16) were evaluated for validity and reliability across age and race-ethnicity groups. Covariates included gender, marital status, education level, cancer site and stage, comorbidities, and functional status. PROMIS physical function short forms showed high internal consistency (Cronbach's α = 0.92-0.96), convergent validity (fatigue, pain interference, FACT physical well-being all r ≥ 0.68), and discriminant validity (unrelated domains all r ≤ 0.3) across survey short forms, age, and race-ethnicity. Known-group differences by demographic, clinical, and functional characteristics performed as hypothesized. Ceiling effects for higher-functioning individuals were identified on most forms. This study provides strong evidence that PROMIS physical function measures are valid and reliable in multiple race-ethnicity and age groups. Researchers selecting specific PROMIS short forms should consider the degree of functional disability in their patient population to ensure that length and content are tailored to limit response burden.


Baohuoside I via mTOR Apoptotic Signaling to Inhibit Glioma Cell Growth.
Baohuoside I, a novel oncotherapeutic agent, has been reported to have anti-cancer effects on a variety of cancers, but its role in glioma and its molecular mechanism are still unclear. The proliferation of U251 cells was detected by real-time cellular analysis (RTCA), CCK-8, Ki67 immunofluorescence and colony formation assay. The effect of Baohuoside I on the invasion and migration of U251 cells was measured by transwell and scratch tests. The apoptosis of U251 cells was detected by flow cytometry. The expression level of related protein was detected by western blotting. Baohuoside I could inhibit the proliferation of human glioma cells and induce apoptosis. Further study showed that the migration and invasion ability of glioma was significantly decreased by Baohuoside I. Western blot revealed the expression of p-AMPKα1 protein was up-regulated, and the expression of p-mTOR and p-S6K was down-regulated after Baohuoside I treatment. Tumorigenesis in nude mice showed that Baohuoside I had an anti-glioma effect in vivo. We propose a natural product, which can inhibit the proliferation, invasion and migration of glioma and may be a valuable anti-tumor candidate. The inhibitory effect of Baohuoside I on the glioma is achieved by inducing the apoptosis of the tumor cells, rather than autophagy. In addition, the pathway to induce cell apoptosis of Baohuoside I is to target the mTOR signal.


Low-carbon economic growth in Chinese cities: a case study in Shenzhen city.
Low-carbon economic growth in cities is important for reduction of carbon emissions in China. As the best practice city in China, Shenzhen city has experienced rapid economic growth with low carbon emissions. The study aims to evaluate the performance of Chinese cities on low-carbon economic growth through the case study of Shenzhen city. The study carries out the Tapio decoupling model for analyzing decoupling state, and uses the Kaya-Logarithmic Mean Divisia Index decomposition model to determine the main driving factors of carbon emissions in Shenzhen. Results indicate that Shenzhen has greatly decoupled carbon emissions with economic growth. The analysis of driving factors of carbon emission shows that the declining energy intensity and the upgrading industrial structure effectively hamper the increase of carbon emissions in Shenzhen. The decline in energy intensity in Shenzhen may come from an improvement of production efficiency of the industries. However, the irrational energy consumption structure, fast-growing economic output, and industry scale are hampering the low carbon emissions of Shenzhen. All estimated industries are highly dependent on coal and oil although some industries have slightly increased their proportion of clean energy consumption. Pursuing more clean energy consumption in the industry will be a key development strategy for reducing emissions in the future. Moreover, as Shenzhen is a fast-growing city, the increasing economic output and industry scale are inevitable. Changing people's way of living could also help in reducing carbon emissions in cities.


Cystic acoustic neuromas: surgical outcome in a series of 58 patients.
We aimed to analyze the clinical characteristics and surgical outcomes of surgically treated cystic acoustic neuromas, and to determine the prognostic significance of the presence of cystic components, by comparing surgical outcomes for solid and cystic tumors. A total of 58 patients (20.4%) with newly diagnosed cystic tumor were studied. The surgical outcome was analyzed in terms of extent of tumor removal, facial nerve preservation, morbidity and mortality; and was compared with that in patients with solid tumors. Facial nerve preservation rates were much better for solid tumors (82.7%) than for cystic tumors (67.9%). The extent of tumor removal was complete in 75.9% of patients with cystic tumors as compared with 90.2% for solid acoustic neuromas. Overall mortality was 8.6%. Cystic tumors differ from solid acoustic neuromas by having a rapid growth pattern, short clinical history and more frequent facial nerve involvement. They are associated with a poorer surgical outcome in terms of facial preservation and overall result.


The effects of time-restricted eating on sleep, cognitive decline, and Alzheimer's disease.
According to the United Nations, by 2050, one in six individuals will be over age 65 globally, and one in four people would be aged 65 and older in western countries. The unprecedented growth of the aging population is associated with increased age-related disorders like Alzheimer's disease (AD) and Mild cognitive impairment (MCI). To date, no cure is known for AD, thus lifestyle interventions including calorie restriction (CR) and time-restricted eating (TRE) are proposed as potential approach to delay the onset and progression of the disease. Sleep disturbances are common in people with MCI and AD. Moreover, accumulating data indicates that pro-inflammatory cytokines including tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), IL-6, IL-8 and IL-10 increase in individuals with AD and MCI versus healthy subjects. Thus, the purpose of the present review is to describe the potential effects of TRE on sleep, cognition decline, and neuroinflammatory markers in humans. Preliminary evidence suggests that TRE may produce neuroprotective effects on cognition and reduce neuroinflammatory markers related to AD in humans. To date, no studies investigated the effects of TRE on sleep disturbances and patients with AD. Thereby, the impact of TRE on cognition in individuals with cognitive decline and AD needs to be investigated further in randomized controlled trials (RCTs).


A fully unsupervised compartment-on-demand platform for precise nanoliter assays of time-dependent steady-state enzyme kinetics and inhibition.
The ability to miniaturize biochemical assays in water-in-oil emulsion droplets allows a massive scale-down of reaction volumes, so that high-throughput experimentation can be performed more economically and more efficiently. Generating such droplets in compartment-on-demand (COD) platforms is the basis for rapid, automated screening of chemical and biological libraries with minimal volume consumption. Herein, we describe the implementation of such a COD platform to perform high precision nanoliter assays. The coupling of a COD platform to a droplet absorbance detection set-up results in a fully automated analytical system. Michaelis-Menten parameters of 4-nitrophenyl glucopyranoside hydrolysis by sweet almond β-glucosidase can be generated based on 24 time-courses taken at different substrate concentrations with a total volume consumption of only 1.4 μL. Importantly, kinetic parameters can be derived in a fully unsupervised manner within 20 min: droplet production (5 min), initial reading of the droplet sequence (5 min), and droplet fusion to initiate the reaction and read-out over time (10 min). Similarly, the inhibition of the enzymatic reaction by conduritol B epoxide and 1-deoxynojirimycin was measured, and Ki values were determined. In both cases, the kinetic parameters obtained in droplets were identical within error to values obtained in titer plates, despite a >10(4)-fold volume reduction, from micro- to nanoliters.


Novel ORL1-selective antagonists with oral bioavailability and brain penetrability.
Following the discovery of 5-chloro-6-[piperazin-1-yl]-1H-benzimidazole as a novel pharmacophore for potent and selective ORL1 antagonist activity, optimization of this new lead by introduction of a methyl substitution on the piperazine ring resulted in a highly potent and selective, orally available, and brain penetrable ORL1 antagonist, 2-(tert-butylthio)-5-chloro-6-[(2R)-4-(2-hydroxyethyl)-2-methylpiperazin-1-yl]-1H-benzimidazole. Stereochemistry of the methyl substituent on the piperazine ring to control the functional activity of other opioid receptors is also described.


Comparison of Near-surface NO2 Pollution with Pandora Total Column NO2 during the Korea-United States Ocean Color (KORUS OC) Campaign.
Near-surface air quality (AQ) observations over coastal waters are scarce, a situation that limits our capacity to monitor pollution events at land-water interfaces. Satellite measurements of total column (TC) nitrogen dioxide (NO2) observations are a useful proxy for combustion sources but the once daily snapshots available from most sensors are insufficient for tracking the diurnal evolution and transport of pollution. Ground-based remote sensors like the Pandora Spectrometer Instrument (PSI) that have been developed to verify space-based total column NO2 and other trace gases are being tested for routine use as certified AQ monitors. The KORUS-OC (Korea-United States Ocean Color) cruise aboard the R/V Onnuri in May-June 2016 represented an opportunity to study AQ near the South Korean coast, a region affected by both local/regional and long-distance pollution sources. Using PSI data in direct-sun mode and in situ sensors for shipboard ozone, CO and NO2, we explore, for the first time, relationships between TC NO2 and surface AQ in this coastal region. Three case studies illustrate the value of the PSI as well as complexities in the surface-column NO2 relationship caused by varying meteorological conditions. Case Study 1 (25-26 May 2016) exhibited a high correlation of surface NO2 to TC NO2 measured by both PSI and Aura's Ozone Monitoring Instrument (OMI) but two other cases displayed poor relationships between in situ and TC NO2 due to decoupling of pollution layers from the surface. With suitable interpretation the PSI TC NO2 measurement demonstrates good potential for working with upcoming geostationary satellites to advance diurnal tracking of pollution.


An electron microscopic cytochemical study on concanavalin A binding sites and their mobility in normal and cystic fibrosis fibroblasts in vitro.
Concanavalin A (Con A) binding sites were visualized ultrastructurally in the cultured fibroblasts from cystic fibrosis patients, obligatory heterozygotes, and normal individuals by peroxidase labeling technique. Con A binding sites were localized as a continuous layer on the external side of the plasma membrane in fixed fibroblasts of the three genotypes. In living cells, Con A induces both lateral and vertical movements of binding sites as expressed by cap formation and internalization of the plasma membrane. Fibroblasts of the three genotypes responded similarly to Con A treatment and failed to show significant detectable differences.


One-pot synthesis and biochemical characterization of a magnetic collagenase nanoflower and evaluation of its biotechnological applications.
Recently, hierarchical magnetic enzyme nanoflowers have been found extensive attention for efficient enzyme immobilization due to high surface area, low mass transfer limitations, active site accessibility, promotion of the enzymatic performance, and facile reusing. Herein, we report the purification of the Bacillus collagenase and then synthesis of magnetic cross-linked collagenase-metal hybrid nanoflowers (mcCNFs). The catalytic efficiency (kcat/Km) value of the immobilized collagenase was 2.2 times more than that of the free collagenase. The collagenase activity of mcCNFs enhanced about 2.9 and 4.6 at 85 and 90 °C, respectively, compared to free collagenase. Thermal stability of mcCNFs increased about 31% and 24% after 3 h of incubation at 50 and 60 °C, respectively. After 10 cycles of reusing, the mCNFs collagenase showed 83% of its initial activity. Results showed that the mcCNFs revealed 1.4 times more activity than the free collagenase in 0.16% protein waste. Furthermore, the hydrolysis value of chicken pie protein wastes by the immobilized enzyme obtained 4 times more than the free collagenase after 240 min incubation at 40 °C. Finally, our results showed that the construction of mcCNFs is an efficient method to increase the enzymatic performance and has excessive potential for the hydrolysis of protein wastes in the food industry.


Oscillatory synchronization model of attention to moving objects.
The world is a dynamic environment hence it is important for the visual system to be able to deploy attention on moving objects and attentively track them. Psychophysical experiments indicate that processes of both attentional enhancement and inhibition are spatially focused on the moving objects; however the mechanisms of these processes are unknown. The studies indicate that the attentional selection of target objects is sustained via a feedforward-feedback loop in the visual cortical hierarchy and only the target objects are represented in attention-related areas. We suggest that feedback from the attention-related areas to early visual areas modulates the activity of neurons; establishes synchronization with respect to a common oscillatory signal for target items via excitatory feedback, and also establishes de-synchronization for distractor items via inhibitory feedback. A two layer computational neural network model with integrate-and-fire neurons is proposed and simulated for simple attentive tracking tasks. Consistent with previous modeling studies, we show that via temporal tagging of neural activity, distractors can be attentively suppressed from propagating to higher levels. However, simulations also suggest attentional enhancement of activity for distractors in the first layer which represents neural substrate dedicated for low level feature processing. Inspired by this enhancement mechanism, we developed a feature based object tracking algorithm with surround processing. Surround processing improved tracking performance by 57% in PETS 2001 dataset, via eliminating target features that are likely to suffer from faulty correspondence assignments.


A cough algorithm for chronic cough in children: a multicenter, randomized controlled study.
The goals of this study were to: (1) determine if management according to a standardized clinical management pathway/algorithm (compared with usual treatment) improves clinical outcomes by 6 weeks; and (2) assess the reliability and validity of a standardized clinical management pathway for chronic cough in children. A total of 272 children (mean ± SD age: 4.5 ± 3.7 years) were enrolled in a pragmatic, multicenter, randomized controlled trial in 5 Australian centers. Children were randomly allocated to 1 of 2 arms: (1) early review and use of cough algorithm ("early-arm"); or (2) usual care until review and use of cough algorithm ("delayed-arm"). The primary outcomes were proportion of children whose cough resolved and cough-specific quality of life scores at week 6. Secondary measures included cough duration postrandomization and the algorithm's reliability, validity, and feasibility. Cough resolution (at week 6) was significantly more likely in the early-arm group compared with the delayed-arm group (absolute risk reduction: 24.7% [95% confidence interval: 13-35]). The difference between cough-specific quality of life scores at week 6 compared with baseline was significantly better in the early-arm group (mean difference between groups: 0.6 [95% confidence interval: 0.29-1.0]). Duration of cough postrandomization was significantly shorter in the early-arm group than in the delayed-arm group (P = .001). The cough algorithm was reliable (κ = 1 in key steps). Feasibility was demonstrated by the algorithm's validity (93%-100%) and efficacy (99.6%). Eighty-five percent of children had etiologies easily diagnosed in primary care. Management of children with chronic cough, in accordance with a standardized algorithm, improves clinical outcomes irrespective of when it is implemented. Further testing of this standardized clinical algorithm in different settings is recommended.


Delaying Invasive Treatment in Unilateral Head and Neck Lymphatic Malformation Improves Outcomes.
Large (De Serres stage [IV-V]) head and neck lymphatic malformations (HNLMs) often have multiple, high-risk, invasive treatments (ITs) to address functional compromise. Logically reducing HNLM ITs should reduce treatment risk. We tested whether delaying HNLM ITs reduces total IT number. Consecutive HNLM patients (n = 199) between 2010 and 2017, aged 0-18 years. ITs (surgery or sclerotherapy) were offered for persistent or dysfunction causing HNLMs. Treatment effectiveness categorized by IT number: optimal (0-1), acceptable (2-5), or suboptimal (>5). Clinical data were summarized, and outcome associations tested (χ2 ). Relative risk (RR) with a Poisson working model tested whether HNLM observation or IT delay (>6 months post-diagnosis) predicts treatment success (i.e., ≤1 IT). Median age at HNLM diagnosis was 1.3 months (interquartile range [IQR] 0-45 m) with 107/199(54%) male. HNLM were stage I-III (174 [88%]), IV-V (25 [13%]). Initial treatment was observation (70 [35%]), invasive (129 [65%]). Treatment outcomes were optimal (137 [69%]), acceptable (36 [18%]), and suboptimal (26 [13%]). Suboptimal outcome associations: EXIT procedure, stage IV-V, oral location, and tracheotomy (p < 0.001). Stage I-III HNLMs were initially observed compared with stage I-III having ITs within 6 months of HNLM diagnosis, had a 82% lower relative treatment failure risk ([i.e., >1 IT], RR = 0.09, 95% CI 0.02-0.36, p < 0.001). Stage I-III HNLMs with non-delayed ITs had reduced treatment failure risk compared with IV-V (RR = 0.47, 95% CI 0.33-0.66, p < 0.001). Observation and delayed IT in stage I-III HNLM ("Grade 1") is safe and reduces IT (i.e., ≤1 IT). Stage IV-V HNLMs ("Grade 2") with early IT have a greater risk of multiple ITs. 4 Laryngoscope, 133:956-962, 2023.


Mapping of the gene coding for the human GM2 activator protein to chromosome 5.
The gene coding for the GM2 activator protein has been mapped to human chromosome 5, using an enzyme-linked immunoadsorbent assay (ELISA) to identify the human protein in human-mouse somatic cell hybrids.


Thrombocytopenia following autologous bone marrow transplantation: evidence for autoimmune aetiology and B cell clonal involvement.
Thrombocytopenia outlasting anaemia and neutropenia is a well recognised sequel of autologous bone marrow transplantation (BMT) but the pathogenesis remains unclear. Autoimmune destruction of platelets has been suggested as a possible mechanism. We studied 5 patients who had undergone autologous BMT and were found to have persistent thrombocytopenia (< 150 x 10(9)/l) 6 months from transplantation with a normal haemoglobin level and granulocyte count. Apart from a mild reduction in the megakaryocyte numbers in one case, no other quantitative or qualitative defects of the megakaryocyte lineage were present to explain the peripheral thrombocytopenia. Two cases had positive anti-platelet autoantibodies. Immunoglobulin heavy chain gene rearrangement studies of peripheral blood and bone marrow mononuclear cells using the polymerase chain reaction showed evidence of clonal rearrangement in one of the two cases with positive anti-platelet autoantibodies. Our results support the previous reports that anti-platelet antibody-mediated destruction of platelets may play a role in the pathogenesis of post-autologous BMT thrombocytopenia. Furthermore, the demonstration of a clonal B cell expansion in one of the cases with anti-platelet antibodies suggests an aetiological link between clonal B cells, autoantibody production and thrombocytopenia.


A novel anticancer effect of butein: inhibition of invasion through the ERK1/2 and NF-kappa B signaling pathways in bladder cancer cells.
There is increasing evidence that epithelial-mesenchymal transition (EMT) plays a critical role in cancer metastasis. Butein is a polyphenolic compound, which has been found to exhibit anti-proliferation effects on cancer cells. Here, we report that in addition to its function as an anti-proliferation agent, butein can inhibit migration and invasion through the ERK1/2 and NF-kappaB signaling pathways in human bladder cancer cells, and this inhibitory effect may be associated with reversal of EMT. These results were further confirmed by RNAi-mediated suppression of NF-kappaB, which partly reverses EMT and inhibits cell invasive ability in vitro. These results suggest a novel function of butein as an invasion inhibitor in bladder cancer.


Managing bibliographic citations using microcomputers.
Programs are now available to construct and retrieve lists of bibliographic citations by microcomputers. Although considerable effort must be expended to learn to use them, once mastered, they can be useful to anyone who must manage large collections of references. There are several ways to use personal computers to store and retrieve bibliographic citations. Word processors can be used to manage relatively small lists of citations. Preprogrammed bibliographic systems are available that are designed specifically for this purpose. A general data base management program can also be adapted for bibliographic purposes and used for other functions as well. This essay has been prepared to provide guidance to those who have a working knowledge of microcomputers and wish to expand this to use a data management system for bibliographic purposes. The dBASE II program is used to illustrate how to set up a bibliographic system. Methods are described on how to prepare citations for storage and retrieval using combinations of key words and Boolean operators, how to prepare selected lists of references and arrange them in alphabetic order or by subject heading, and how to print tailored lists of citations. The system was found to be highly responsive to commands and able to provide rapid retrieval of information.


Heterogeneity and complexity of the prostate epithelium: New findings from single-cell RNA sequencing studies.
The recent advent of single-cell RNA-sequencing technology has provided new fundamental insights into the heterogeneity of the prostate epithelium. Several independent studies have described extensive heterogeneity of the luminal epithelial compartment, including a major division between a novel population of luminal cells located in the proximal region of the prostate ducts versus luminal cells located more distally. Proximal luminal cells as well as novel periurethral cells display increased progenitor potential in organoid culture and tissue reconstitution assays, but not in lineage-tracing analyses during prostate homeostasis, suggesting context-dependent plasticity of these populations. Here we describe and synthesize recent findings regarding the epithelial cell populations in the mouse prostate, draw comparisons to the human prostate, and address the relevance of these findings to prostate diseases and cancer.


CpG island methylator phenotype (CIMP) in cancer: causes and implications.
Strong evidence exists for a subgroup of tumours, from a variety of tissue types, exhibiting concordant tumour specific DNA methylation: the "CpG island methylator phenotype" (CIMP). Occurrence of CIMP is associated with a range of genetic and environmental factors, although the molecular causes are not well-understood. Both increased expression and aberrant targeting of DNA methyltransferases (DNMTs) could contribute to the occurrence of CIMP. One under-explored area is the possibility that DNA damage may induce or select for CIMP during carcinogenesis or treatment of tumours with chemotherapy. DNA damaging agents can induce DNA damage at guanine rich regions throughout the genome, including CpG islands. This DNA damage can result in stalled DNA synthesis, which will lead to localised increased DNMT1 concentration and therefore potentially increased DNA methylation at these sites. Chemotherapy can select for cells which have increased tolerance to DNA damage due to increased lesion bypass, in some cases by mechanisms which involve inactivation of genes by CpG island methylation. CIMP has been associated with worse patient prognosis, probably due to increased epigenetic plasticity. Therefore, further clinical testing of the diagnostic and prognostic value of the current CIMP markers, as well as increasing our understanding of the molecular causes underlying CIMP are required.


[Surgical treatment of adenocarcinoma of the esophagogastric junction type II : Comparison between transhiatal extended gastrectomy and thoracoabdominal esophagectomy].
The incidence of adenocarcinoma of the esophagogastric junction (AEG) has significantly increased. However, the best surgical treatment for AEG type II is still the subject of current research. The goal of this retrospective cohort study is to compare survival and recurrence rates in patients, who underwent either thoracoabdominal esophagectomy (TAE) or transhiatal extended gastrectomy (TEG). The study is based on a cohort of 272 patients diagnosed with AEG type II between 2002 and 2020, recorded by a population-based clinical cancer registry. Of the included patients 63 underwent TAE and 209 TEG. In order to compare overall survival, recurrence rates, and recurrence free survival, we applied the Kaplan-Meier method, univariable and multivariable Cox regression. Our analysis showed no statistically significant difference concerning overall survival (p = 0.333). However a tendency towards higher survival rates after TAE for the period from 2016-2020 (p = 0,058) is possible. In contrast a significant difference concerning higher cumulative recurrence rates after TAE was found after Kaplan-Meier analysis (p = 0.049). This trend was not observed for the time after 2016 (p = 0.993), in which over 50% of TAE were performed. No differences were found regarding recurrence-free survival (p = 0.772). Our findings in a rather small cohort are concordant with most studies showing no differences or a trend towards better survival after TAE. Other studies found no significant difference regarding recurrence-free survival as well. In conclusion, no significant differences were found between TEG and TAE in surgical treatment of AEG type II.


Siloxyaluminate and Siloxygallate Complexes as Models for Framework and Partially Hydrolyzed Framework Sites in Zeolites and Zeotypes.
Anionic molecular models for nonhydrolyzed and partially hydrolyzed aluminum and gallium framework sites on silica, M[OSi(OtBu)3 ]4 - and HOM[OSi(OtBu)3 ]3 - (where M=Al or Ga), were synthesized from anionic chlorides Li{M[OSi(OtBu)3 ]3 Cl} in salt metathesis reactions. Sequestration of lithium cations with [12]crown-4 afforded charge-separated ion pairs composed of monomeric anions M[OSi(OtBu)3 ]4 - with outer-sphere [([12]crown-4)2 Li]+ cations, and hydroxides {HOM[OSi(OtBu)3 ]3 } with pendant [([12]crown-4)Li]+ cations. These molecular models were characterized by single-crystal X-ray diffraction, vibrational spectroscopy, mass spectrometry and NMR spectroscopy. Upon treatment of monomeric [([12]crown-4)Li]{HOM[OSi(OtBu)3 ]3 } complexes with benzyl alcohol, benzyloxide complexes were formed, modeling a possible pathway for the formation of active sites for Meerwin-Ponndorf-Verley (MPV) transfer hydrogenations with Al/Ga-doped silica catalysts.


Identification and characterization of Ageratum yellow vein Malaysia virus (AYVMV) and an associated betasatellite among begomoviruses infecting Solanum lycopersicum in Malaysia.
The study describes results of a survey of tomato fields for the presence of begomoviruses from different regions of Peninsular Malaysia. An ORF-based (C2 and C3) study was performed to determine the distribution of begomoviruses associated with a severe leaf curl disease in tomato-growing areas of Peninsular Malaysia. Viral DNA was isolated from symptomatic tomato plants, and begomovirus association was confirmed by PCR using DNA-A degenerate primers. The C2 and C3 sequences of the putative begomoviruses were similar to two corresponded ORFs of different geographically separated strains of begomoviruses: Pepper yellow leaf curl Indonesia virus and Tomato yellow leaf curl Kanchanaburi virus. The present study also identified a unique isolate, Ageratum yellow vein Malaysia virus (AYVMV) among above mentioned survey. It has a single-stranded DNA component and its associated betasatellite. The single-stranded DNA component is consisting of 2750 nt with six open reading frames and an organization resembling that of monopartite geminiviruses. The full length of viral single-stranded DNA component genome obtained using next generation sequencing (NGS) showed the highest sequence identity (99%) with Ageratum yellow vein virus (AYVV-BA). The betasatellite component genome obtained by NGS has 1342 nt and showed the highest sequence identity (91%) with the Pepper yellow leaf curl betasatellite. Following ICTV guidelines, Ageratum yellow vein Malaysia virus was assigned the abbreviation AYVMV with sequence and phylogenetic analysis indicating that it might have evolved by recombination of two or more viral ancestors.


[Primary carcinoma of the gallbladder: ultrasonic features (remarks on 8 cases)].
The authors illustrate the ultrasound features of gallbladder carcinoma after reviewing literature and basing upon their own experience. The diagnosis of this neoplasia is formulated more frequently today by introduction of scanning means, but it remains very difficult on initial phase because of its precocious lymphatic and hilar infiltration.


[Psychologic adjustment process following high cervical spine paralysis and its support by a bedside reading apparatus].
The process of psychological adjustment, which has to go on after high levelled lesions of the cervical spinal cord, passes through several phases. It needs a lot of empathy, plenty of time for confidential talks and psychological knowledge as well as an individualizing cooperation between the several levels and stages of combined treatment during rehabilitation. We could give this process a considerable advance by a (half-) automatic bedside-projection of belletristic literature, which can be continuously read on a screen from long film strips by means of a modified Pentacon system. The introduction of this new object in life proved a good support for the threatened personality of those badly handicapped patients.


Correlation of the inhomogeneous field as a measure of sample heterogeneity.
In this paper we present a method to determine the pore and grain size heterogeneity using a correlation between the transverse relaxation time and the susceptibility induced inhomogeneous magnetic field. By using a CPMG echo train, we are able to correlate T(2) with the inhomogeneous internal magnetic field, B(z)(i). We first introduce a simple simulation to study the correlation between the internal magnetic field gradient, ∣g∣, and B(z)(i), where this correlation is analogous to the T(2)-B(z)(i) correlation with horizontal inversion. We then acquire experimental data from two samples, one a simple bead pack, and the other a more complicated sandstone rock core. We find that indeed this method can indicate the presence of pore and grain size heterogeneities in the sample.


Dihydrotestosterone inhibits insulin-stimulated cyclin D2 messenger ribonucleic acid expression in rat ovarian granulosa cells by reducing the phosphorylation of insulin receptor substrate-1.
The effect of 5alpha-dihydrotestosterone (DHT) on insulin-stimulated granulosa cell proliferation was examined using cyclin D2 mRNA as a marker. Granulosa cells from 3-d estradiol-treated immature rats showed a concentration-dependent increase in cyclin D2 mRNA expression in response to insulin. Exposure to DHT reduced the insulin-stimulated cyclin D2 mRNA expression. Inhibition of the two insulin-signaling pathways, ERK and phosphatidylinositol 3 kinase (PI3 kinase), by using specific inhibitors, also reduced this insulin-stimulated response. These results suggest that both ERK and PI3 kinase signaling are involved in insulin stimulated granulosa cell proliferation. DHT exposure resulted in reduced insulin-stimulated ERK phosphorylation. DHT treatment also reduced the insulin mediated insulin receptor substrate-1 and Raf-1 phosphorylation, the upstream molecules of ERK in insulin signaling pathway. Additionally, inhibition of insulin stimulated PI3 kinase activation reduced ERK phosphorylation. The present study therefore shows that the inhibitory effect of DHT on insulin-stimulated granulosa cell proliferation occurs early in the signaling pathway at the level of insulin receptor substrate-1 phosphorylation, leading to reduced ERK phosphorylation and subsequent inhibition of cyclin D2 mRNA expression.


L-tyrosine administration increases acetylcholinesterase activity in rats.
Tyrosinemia is a rare genetic disease caused by mutations on genes that codify enzymes responsible for tyrosine metabolism. Considering that tyrosinemics patients usually present symptoms associated with central nervous system alterations that ranges from slight decreases in intelligence to severe mental retardation, we decided to investigate whether acute and chronic administration of L-tyrosine in rats would affect acetylcholinesterase mRNA expression and enzymatic activity during their development. In our acute protocol, Wistar rats (10 and 30 days old) were killed one hour after a single intraperitoneal L-tyrosine injection (500 mg/kg) or saline. Chronic administration consisted of L-tyrosine (500 mg/kg) or saline injections 12 h apart for 24 days in Wistar rats (7 days old) and rats were killed 12 h after last injection. Acetylcholinesterase activity was measured by Ellman's method and acetylcholinesterase expression was carried out by a semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) assay. We observed that acute (10 and 30 days old rats) and chronic L-tyrosine administration increased acetylcholinesterase activity in serum and all tested brain areas (hippocampus, striatum and cerebral cortex) when compared to control group. Moreover, there was a significant decrease in mRNA levels of acetylcholinesterase in hippocampus was observed after acute protocol (10 and 30 days old rats) and in striatum after chronic protocol. In case these alterations also occur in the brain of the patients, our results may explain, at least in part, the neurological sequelae associated with high plasma concentrations of tyrosine seen in patients affected by tyrosinemia type II.


Flavor characteristics of Shanlan rice wines fermented for different time based on HS-SPME-GC-MS-O, HS-GC-IMS, and electronic sensory analyses.
Flavor characteristics of Shanlan rice wines with different fermentation time were analyzed. Results suggested that 3-methyl-1-butanol, phenylethyl alcohol, ethyl acetate, and diethyl succinate were the characteristic volatiles of Shanlan rice wine by using headspace solid-phase microextraction-gas chromatography-mass spectrometry-olfactometry. The most varieties (38) of volatiles appeared at a 3-year-fermentation time, contributing a unique and harmonious aroma to the Shanlan rice wine fermented for 3 years, but only 19 types were observed at 45-days and 1-year fermentation times. A similar trend was intuitively visible in the headspace-gas chromatography-ion mobility spectrometry analysis. The Shanlan rice wine fermented for 3 years had a similar taste profile to that fermented for 45 days, but with distinguishing contents of free amino acids (1352.80 mg/L and 2261.50 mg/L, respectively) and organic acids (9.58 g/L and 49.88 g/L, respectively). The Shanlan rice wine fermented for 1 year had a strong taste with more intensity of most taste attributes.


Salivary immunoglobulins in diseases affecting salivary glands.
Inflammatory disorders of the salivary glands cause marked abnormalities in secretion of immunoglobulins. The changes are reversible, however, in a relatively short period of time. More subtle changes in immunoglobulin transport are present in such diseases as Sjogren's syndrome and diabetes. No changes are discernable in alcoholic cirrhosis. Apparently salivary gland basement membranes are much more resistant to derangement than plasma membranes and the secretory IgA system can continue to operate in the face of numerous affronts. If nothing else these findings suggest that vaccination procedures in the region of the salivary glands may produce an inflammatory response, but it would be readily reversible. In addition, one could anticipate a functioning s-IgA system even in salivary glands with alterations in electrolyte transport. It is difficult to anticipate the situation in immunologically compromised patients, such as those on hemodialysis. Fortunately these patients represent a small population and for them at least, caries is a relatively minor concern.


Assessment of bleeding risk in low-weight patients receiving prophylactic subcutaneous unfractionated heparin.
Underweight patients may be at an increased risk of bleeding while receiving venous thromboembolism (VTE) prophylaxis. Additional evidence is needed to identify patient-specific factors associated with bleeding. The objective of the study was to describe the incidence and identify risk factors associated with bleeding in low-weight (⩽ 60 kg) adult patients receiving subcutaneous unfractionated heparin (SQH) for VTE prophylaxis. This was a single-center, retrospective, nested case-control study of low-weight patients receiving SQH for VTE prophylaxis for ⩾ 48 hours. Cases, patients with clinically relevant bleeding while receiving SQH, and controls, patients without a bleeding event, were matched in a 1:3 ratio for age, sex, primary service (surgical or medical), and time at risk of bleeding on SQH to determine factors associated with bleeding. A total of 3761 patients met the inclusion criteria, of which 38 cases of clinically relevant bleeding were identified. The bleeding incidence was 1% at hospital day 6 and 2.8% at hospital day 14. Most patients in this study (69%) received SQH 5000 units three times daily. ICU admission at SQH start was associated with bleeding, OR 2.97 (95% CI 1.21-7.29). Bleeding in low-weight patients on prophylactic SQH was uncommon. Patients admitted to the ICU at time of SQH start may be at a higher risk of bleeding. Further studies are needed to detect additional risk factors associated with bleeding and investigate the effects of reduced dosing in this population.


Changes in weekly working hours, proportion of doctors with hours above the limitations of European Working Time Directive (EWTD) and time spent on direct patient care for doctors in Norway from 2016 to 2019: a study based on repeated surveys.
To compare the total weekly working hours, proportions with work hours above the limitations of European working time directive (EWTD) and time spent on direct patient care in 2016 and 2019 for doctors working in different job positions in Norway. Repeated postal surveys in 2016 and 2019. Norway. Representative samples of doctors; the response rates were 73.1% (1604/2195) in 2016 and 72.5% (1511/2084) in 2019. Self-reported weekly working hours, proportions with hours above the limitations of EWTD defined as >48 hours/week and time spent on direct patient care. Linear mixed models with estimated marginal means and proportions. From 2016 to 2019, the weekly working hours increased significantly for male general practitioners (GPs) (48.7 hours to 50.9 hours) and male hospital doctors in leading positions (48.2 hours to 50.5 hours), and significantly decreased for female specialists in private practice (48.6 hours to 44.9 hours). The proportion of time spent on direct patient care was noted to be similar between genders and over time. In 2019, it was higher for specialists in private practice (66.4%) and GPs (65.5%) than for doctors in other positions, such as senior hospital consultants (43.5%), specialty registrars (39.8%) and hospital doctors in leading positions (34.3%). Working >48 hours/week increased significantly for both male and female GPs (m: 45.2% to 57.7%; f: 27.8% to 47.0%) and hospital doctors in leading positions (m: 34.4% to 57.1%; f: 17.4% to 46.4%), while it significantly decreased for female specialty registrars (13.2% to 6.9%). Working hours increased significantly for GPs and hospital doctors in leading positions from 2016 to 2019, resulting in increased proportions of doctors with work hours above the EWTD. As work hours above the EWTD can be harmful for health personnel and for safety at work, initiatives to reduce long working weeks are needed.


Dynamic membrane-assisted fermentation of food wastes for enhancing lactic acid production.
A dynamic membrane (DM) module was inserted into a fermentation reactor to separate soluble products from the fermented mixture to increase lactic acid (LA) production from food wastes under acidogenic conditions (uncontrolled pH, pH 4 and 5). With a high total suspended solid content (20-40g/L) in the fermenter, a stable DM could be maintained through regular backwashing. By effectively intercepting suspended solids and lactic acid bacteria (LAB), the fermenter was able to increase microbial activity and largely promote LA yield. Hydrolysis and acidogenesis rates increased with pH, and the highest LA yield (as high as 0.57g/g-TS) was obtained at pH 4. The microbial community analysis showed that the relative abundance of Lactobacillus increased to 96.4% at pH 4, but decreased to 43.3% at pH 5. In addition, the DM could be easily recovered by intercepting larger particles in less than 2h after each cycle of periodic backwashing.


Synergic Interplay Between Halogen Bonding and Hydrogen Bonding in the Activation of a Neutral Substrate in a Nanoconfined Space.
The principle of amplified halogen bonding (XB) in a small space is exploited as a catalytic tool for the activation of an XB acceptor substrate in a nanoconfined environment. The inner cavity of the resorcinarene capsule has been equipped with an XB catalyst bearing an ammonium unit acting as a Trojan horse to drive the catalyst inside the capsule. In the presence of a specific XB catalyst, the capsule is able to catalyze a Michael reaction between N-methylpyrrole and methyl vinyl ketone. In the bulk medium in absence of the resorcinarene capsule, the XB catalyst is catalytically ineffective. Quantum-mechanical investigations highlight that the Michael reaction proceeds through the activation of the carbonyl group by synergistically enhanced halogen/hydrogen-bonding interactions and takes place in an open pentameric capsule.


Altered gene expression in busulfan-resistant human myeloid leukemia.
Busulfan (Bu) resistance is a major obstacle to hematopoietic stem cell transplantation (HSCT) of patients with chronic or acute myelogenous leukemia (CML or AML). We used gene expression analysis to identify cellular factors underlying Bu resistance. Two Bu-resistant leukemia cell lines were established, characterized and analyzed for differentially expressed genes. The CML B5/Bu250(6) cells are 4.5-fold more resistant to Bu than their parental B5 cells. The AML KBM3/Bu250(6) cells are 4.0-fold more Bu-resistant than KBM3 parental cells. Both resistant sublines evade Bu-mediated G2-arrest and apoptosis with altered regulations of CHK2 and CDC2 proteins, constitutively up-regulated anti-apoptotic genes (BCL-X(L), BCL2, BCL2L10, BAG3 and IAP2/BIRC3) and down-regulated pro-apoptotic genes (BIK, BNIP3, and LTBR). Bu-induced apoptosis is partly mediated by activation of caspases; use of the inhibitor Z-VAD-FMK completely abrogated PARP1 cleavage and reduced apoptosis by approximately 50%. Furthermore, Bu resistance in these cells may be attributed in part to up-regulation of HSP90 protein and activation of STAT3. The inhibition of HSP90 with geldanamycin attenuated phosphorylated STAT3 and made B5/Bu250(6) and KBM3/Bu250(6) more Bu-sensitive. The analysis of cells derived from patients classified as either clinically resistant or sensitive to high-dose Bu-based chemotherapy indicated alterations in gene expression that were analogous to those observed in the in vitro model cell lines, confirming the potential clinical relevance of this model for Bu resistance.


Potentiality of Bacillus subtilis as biocontrol agent for management of anthracnose disease of chilli caused by Colletotrichum gloeosporioides OGC1.
A soil bacterium, Bacillus subtilis, isolated from the rhizosphere of Chilli, showed high antagonistic activity against Colletotrichum gloeosporioides OGC1. A clear inhibition zone of 0.5-1 cm was observed in dual plate assay. Microscopic observations showed a clear hyphal lysis and degradation of fungal cell wall. In dual liquid cultures, the B. subtilis strain inhibited the C. gloeosporioides up to 100 % in terms of dry weight. This strain also produced a clear halo region on chitin agar medium plates containing 0.5 % colloidal chitin, indicating that it excretes chitinase. The strain also produced other mycolytic enzymes-glucanase and cellulase, demonstrated by a clear zone of hydrolysis of yeast cell wall glucan (YCW 0.1 % v/v) and carboxymethylcellulose (CMC 0.1 % v/v). In liquid cultures, the strain showed appreciable levels of chitinase, glucanase and cellulase activities and hydrolytic activity with C. gloeosporioides OGC1 mycelia as the substrate. The role of the B. subtilis strain in suppressing the fungal growth in vitro was studied in comparison with a UV mutant of that strain, which lacked both antagonistic and hydrolytic activity. The mycolytic enzyme mediated antagonism of B. subtilis was further demonstrated by heat inactivation (70-100 °C), treatment with trypsin and TCA of the crude enzyme extract which lacked antifungal property also. Treatment of the chilli seeds with Bacillus sp. culture showed 100 % germination index similar to the untreated seeds. The treatment of the seed with co-inoculation of the pathogen with Bacillus sp. culture showed 65 % reduction in disease incidence by the treatment as compared to the seed treated with pathogen alone (77.5 %).


Novel niclosamide-derived adjuvants elevating the efficacy of polymyxin B against MDR Pseudomonas aeruginosa DK2.
Pseudomonas aeruginosa (P. aeruginosa) DK2 is a multidrug-resistant (MDR) gram-negative bacterial pathogen, being observed serious resistance to the 'last-resort' antibiotic, polymyxin B (PB). Combination therapies with adjuvants have emerged as effective strategies to reactivate the antibiotics resisted by MDR bacteria. Herein, we screened a library of approved drugs and found that niclosamide (NIC), an anthelmintic drug, could potentiate the efficacy of PB against MDR P. aeruginosa DK2. Next, a series of novel NIC-derived adjuvants were designed, synthesized, and evaluated the synergistic activity with PB. Among them, the combination of 15 with PB displayed superior elimination of P. aeruginosa DK2 in vitro and in vivo compared with the single administration. Moreover, this combination decelerated PB-resistance progress in DK2, along with lower potential toxicity. Overall, this study provides a strategy for development antibiotic adjuvants to potentiate PB against MDR P. aeruginosa infections.


The future excess fraction of cancer due to lifestyle factors in Australia.
Many cancers are caused by exposure to lifestyle, environmental, and occupational factors. Earlier studies have estimated the number of cancers occurring in a single year which are attributable to past exposures to these factors. However, there is now increasing appreciation that estimates of the future burden of cancer may be more useful for policy and prevention. We aimed to calculate the future number of cancers expected to arise as a result of exposure to 23 modifiable risk factors. We used the future excess fraction (FEF) method to estimate the lifetime burden of cancer (2016-2098) among Australian adults who were exposed to modifiable lifestyle, environmental, and occupational risk factors in 2016. Calculations were conducted for 26 cancer sites and 78 cancer-risk factor pairings. The cohort of 18.8 million adult Australians in 2016 will develop an estimated 7.6 million cancers during their lifetime, of which 1.8 million (24%) will be attributable to exposure to modifiable risk factors. Cancer sites with the highest number of future attributable cancers were colon and rectum (n = 717,700), lung (n = 380,400), and liver (n = 103,200). The highest number of future cancers will be attributable to exposure to tobacco smoke (n = 583,500), followed by overweight/obesity (n = 333,100) and alcohol consumption (n = 249,700). A significant proportion of future cancers will result from recent levels of exposure to modifiable risk factors. Our results provide direct, pertinent information to help determine where preventive measures could best be targeted.


The pathophysiology of faecal spotting in obese subjects during treatment with orlistat.
The intermittent loss of oil or liquid faeces ('spotting') is an adverse effect that occurs in obese patients during treatment with the lipase inhibitor orlistat; the pathophysiology is unknown. To investigate the effects of orlistat on anorectal sensorimotor function and continence. Obese subjects susceptible to spotting were identified by an unblind trial of orlistat. Obese spotters (n = 15) and non-spotters (n = 16) completed a randomized, double-blind, cross-over trial of orlistat and placebo. Anorectal function was assessed by rectal barostat and anal manometry, together with a novel stool substitute retention test, a quantitative measurement of faecal continence. Orlistat increased stool volume and raised faecal fat and water. Treatment had no effect on anorectal motor function, but rectal sensation was reduced; on retention testing, the volume retained was increased. Subjects susceptible to spotting had lower rectal compliance, heightened rectal sensitivity and weaker resting sphincter pressure than non-spotters. On retention testing, gross continence was maintained; however, spotters lost small volumes of rectal contents during rectal filling. Treatment with orlistat has no direct adverse effects on anorectal function or continence. Spotting occurs during treatment with orlistat when patients with sub-clinical anorectal dysfunction are exposed to increased stool volume and altered stool composition.


Molecular analysis of Drosophila glutamate receptors.
Insects and other invertebrates use L-glutamate as a neurotransmitter in the central nervous system and at the neuromuscular junction. In contrast to the well-studied effects of L-glutamate on invertebrate muscle cells, relatively little is known about the physiological role of glutamate receptors (GluRs) in the invertebrate central nervous system. We have applied a molecular cloning approach to elucidate the molecular structure of neuronal and muscle-specific Drosophila glutamate receptor subunits (DGluRs). Several domains conserved between rat GluR subunits and DGluRs indicate regions of high functional significance. Drosophila genetics may now be used as a valuable experimental tool to gain further insight into the role of DGluRs in development, synaptic plasticity and control of gene expression.


CAG (cytarabine, aclarubicin and granulocyte colony-stimulating factor) regimen for core binding factor acute myeloid leukaemia with measurable residual disease.
Acute myeloid leukaemia (AML) with t (8;21) or inv (16), called core binding factor (CBF) AML, has a favourable prognosis. However, some CBF-AML patients have persistent measurable residual disease (MRD) and are more likely to relapse after standard chemotherapy treatment. The CAG regimen, composed of cytarabine, aclarubicin and granulocyte colony-stimulating factor, has been proven to be effective and safe in treating refractory AML patients. We performed a retrospective study to evaluate the efficacy of the CAG regimen to eliminate MRD detected by RUNX1::RUNX1T1 and CBFβ::MYH11 transcript levels by quantitative polymerase chain reaction (Q-PCR) among 23 patients. Molecular response was defined as the ratio of fusion transcript after treatment to that before treatment less than or equal to 0.5. The molecular response rate and median decrease ratio of fusion transcripts at the molecular level of the CAG regimen were 52% and 0.53, respectively. The median fusion transcripts before CAG treatment was 0.25% whereas after CAG was 0.11%. Among the 15 patients who had a poor molecular response to the high/intermediate-dose cytarabine regimen, the median decrease ratios of transcripts at the molecular level of high/intermediate-dose cytarabine and CAG were 1.55 and 0.53 (P = 0.028), respectively, and 6 of 15 patients achieved a molecular response to CAG (40%). The median disease-free survival was 18 months, and the overall survival rate at 3 years among all patients was 72.7% ± 10.7%. The common grades 3-4 adverse events were nausea (100%), thrombocytopenia (39%) and neutropenia (37.5%). The CAG regimen may have activity in CBF-AML patients and could provide a new option for patients who have a poor molecular response to high/intermediate-dose cytarabine.


Bladder cancer stage and mortality: urban vs. rural residency.
Relative to urban populations, rural patients may have more limited access to care, which may undermine timely bladder cancer (BCa) diagnosis and even survival. We tested the effect of residency status (rural areas [RA < 2500 inhabitants] vs. urban clusters [UC ≥ 2500 inhabitants] vs. urbanized areas [UA, ≥50,000 inhabitants]) on BCa stage at presentation, as well as on cancer-specific mortality (CSM) and other cause mortality (OCM), according to the US Census Bureau definition. Multivariate competing risks regression (CRR) models were fitted after matching of RA or UC with UA in stage-stratified analyses. Of 222,330 patients, 3496 (1.6%) resided in RA, 25,462 (11.5%) in UC and 193,372 (87%) in UA. Age, tumor stage, radical cystectomy rates or chemotherapy use were comparable between RA, UC and UA (all p > 0.05). At 10 years, RA was associated with highest OCM followed by UC and UA (30.9% vs. 27.7% vs. 25.6%, p < 0.01). Similarly, CSM was also marginally higher in RA or UC vs. UA (20.0% vs. 20.1% vs. 18.8%, p = 0.01). In stage-stratified, fully matched CRR analyses, increased OCM and CSM only applied to stage T1 BCa patients. We did not observe meaningful differences in access to treatment or stage distribution, according to residency status. However, RA and to a lesser extent UC residency status, were associated with higher OCM and marginally higher CSM in T1N0M0 patients. This observation should be further validated or refuted in additional epidemiological investigations.


Ferritin protein nanocage ion channels: gating by N-terminal extensions.
Ferritin protein nanocages, self-assembled from four-α-helix bundle subunits, use Fe(2+) and oxygen to synthesize encapsulated, ferric oxide minerals. Ferritin minerals are iron concentrates stored for cell growth. Ferritins are also antioxidants, scavenging Fenton chemistry reactants. Channels for iron entry and exit consist of helical hairpin segments surrounding the 3-fold symmetry axes of the ferritin nanocages. We now report structural differences caused by amino acid substitutions in the Fe(2+) ion entry and exit channels and at the cytoplasmic pores, from high resolution (1.3-1.8 Å) protein crystal structures of the eukaryotic model ferritin, frog M. Mutations that eliminate conserved ionic or hydrophobic interactions between Arg-72 and Asp-122 and between Leu-110 and Leu-134 increase flexibility in the ion channels, cytoplasmic pores, and/or the N-terminal extensions of the helix bundles. Decreased ion binding in the channels and changes in ordered water are also observed. Protein structural changes coincide with increased Fe(2+) exit from dissolved, ferric minerals inside ferritin protein cages; Fe(2+) exit from ferritin cages depends on a complex, surface-limited process to reduce and dissolve the ferric mineral. High concentrations of bovine serum albumin or lysozyme (protein crowders) to mimic the cytoplasm restored Fe(2+) exit in the variants to wild type. The data suggest that fluctuations in pore structure control gating. The newly identified role of the ferritin subunit N-terminal extensions in gating Fe(2+) exit from the cytoplasmic pores strengthens the structural and functional analogies between ferritin ion channels in the water-soluble protein assembly and membrane protein ion channels gated by cytoplasmic N-terminal peptides.


Outcomes following transcatheter repair in patients with functional mitral regurgitation not receiving guideline directed medical therapy in Israel.
Transcatheter edge to edge repair (TEER) improves prognosis in patients with functional mitral regurgitation (FMR) receiving guideline directed medical therapy (GDMT). Many patients with FMR do not receive GDMT and the utility of TEER in this population remains unclear. We retrospectively studied patients undergoing TEER. Clinical, echocardiographic and procedural variables were recorded. GDMT was defined as use of RAAS inhibitors and MRAs unless GFR was under 30 as well as beta blockers. The primary endpoint of the study was one year mortality. 168 patients (mean age 71.3 ± 9.3; 66% males) with FMR who underwent TEER were included of whom 116 (69%) received GDMT at the time of TEER and 52 (31%) did not. There were no significant demographic or clinical differences between the groups. There were no significant differences in procedural success and complications between groups. One year mortality was identical in the two groups (15% vs. 15%; RR 1.06, CI 0.43-2.63, P = 0.90). Our findings suggest that procedural success and one year mortality following TEER was not significantly different in HFREF patients with FMR with or without GDMT. Larger, prospective studies are necessary to define the benefit of TEER in this population.


Basis of neurotropism of rabies virus: a new hypothesis.
There is evidence that rabies viruses travel along the peripheral nerves, in the axoplasm to reach the central nervous system (CNS). It is hypothesised that rabies viruses are attracted electrically or magnetically by the nerve impulses to the CNS. This explains why rabies viruses show specific affinity towards nerve tissue. The present hypothesis offers a new approach to the study of pathogenesis of the various neurotropic viruses.


Bioethics and power: Informed consent procedures in post-socialist Latvia.
This paper explores two lines of development in the donor consent procedures in post-Soviet Latvia. The paper is based on secondary analysis of interview, focus group discussion data, and media and legal text material collected throughout three previously conducted research projects on organ transplantation, population genome project and xenotransplantation focusing on the historical development of the issues of donor consent across these three fields of medical technologies. The paper argues that the quality of consent depends not as much on political and legal change per se as on the strengthening of the position of both medical specialists and donors, facilitating bonds between the two.


An efficient expression vector for stable expression in human liver cells.
The expression of the bacterial neomycin resistance (NmR)-encoding gene was examined under control of the promoter region of the gene encoding the human polypeptide chain elongation factor 1 alpha (EF-1 alpha). In a human liver cell line, HepG2, the plasmid pEF321-neo directed higher expression of the bacterial neo gene than the NmR gene expression vectors using other promoters, like the SV40 early region or thymidine kinase of Herpes simplex virus and SV40 early region promoter linked to human T-cell leukemia virus-1 enhancer sequences.


Diagnostic value of liquid-based cytology test in intrathoracic lymph nodes and lung lesions sampled by endobronchial ultrasonography-transbronchial needle aspiration.
Liquid-based cytology (LBC) has begun to be used in non-gynecological samples such as endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA). This study aims to investigate the diagnostic value of LBC in intrathoracic lymph nodes and lung lesions sampled with EBUS-TBNA. In total, 174 cases that underwent EBUS-TBNA between July 2020 and February 2021 were included (75 and 99 cases were prepared using conventional and LBC methods, respectively). The two groups were compared in terms of diagnostic categories, number of slides, cell blocks, slides per location, locations sampled, immunohistochemical studies, sensitivity, specificity, and diagnostic accuracy. The percentages of malignant, suspicious for malignancy, benign, and non-diagnostic (ND) cases were 51.8%, 1.1%, 39.6%, and 7.5%, respectively. The LBC and conventional group (CG) had similar rates in the diagnostic categories, except for ND (3.0% and 13.3%, respectively). The sensitivity of LBC and CG were 90.4% and 85.7%, respectively. There were no differences in the specificity and diagnostic accuracy between groups. There was a statistically significant difference between groups in terms of the number of slides, number of slides per location, number of cell blocks, and locations sampled (p < .001, p < .001, p < .05, p < .05). The LBC technique can be used for samples taken with EBUS-TBNA. Rapid fixation and the absence of artificial problems greatly reduce the rate of ND samples in LBC slides. Other important advantages are a lower number of slides to examine and a greater number of cell blocks.


[Ultraviolet A radiation and the skin. Implications of activated forms of oxygen. Current trends and newest results].
Involvement of activated oxygen species in the responses of skin to solar ultraviolet radiations, especially ultraviolet A radiations, is being addressed by an increasing number of studies. The aim of this review is to outline the concept of "photooxidative stress". The various activated oxygen species, mechanisms involved in their formation, potential cellular targets, and cell defense mechanisms are discussed. Recently published findings are briefly described to illustrate this fast developing line of research.


Abdominal neuroendocrine neoplasm with misplaced exocytosis.
A 22-year-old man presented with epigastric fullness and tenderness. A 14-cm mass that was adherent to the sigmoid colon was removed surgically. The tumor exhibited ultrastructural evidence of neuroendocrine differentiation but defied precise classification. The tumor exhibited misplaced exocytosis, a finding not previously described in neuroendocrine tumors other than prolactin cell adenomas.