Effects of high doses of glucocorticoids on free amino acids, ribosomes and protein turnover in human muscle.
Treatment with glucocorticosteroids causes a negative nitrogen balance, but the kinetic mechanisms responsible for this catabolic effect are controversial. We investigated the effects of 60 mg day(-1) prednisolone on protein synthesis and degradation in human skeletal muscle. Healthy adults (n = 9) were studied in the postabsorptive state, before and after 3 days of prednisolone treatment. The L-[ring 2,6(-3)H(5)]-phenylalanine tracer technique, concentration and size distribution of the ribosomes, mRNA content of the ubiquitin-proteasome pathway components in muscle, phenylalanine flux across the leg, and the free amino acid concentrations in skeletal muscle were used to study muscle protein metabolism. The concentrations of most amino acids in arterial blood increased after prednisolone. There were also increased effluxes of phenylalanine, asparagine, arginine, alanine, methionine and isoleucine from the leg. The rate of protein degradation, as measured by the appearance rate (Ra) of phenylalanine, increased by 67% (P = 0.023) which, together with a doubling of the net release of phenylalanine from the leg (P = 0.007), indicated accelerated protein degradation. The pathway was not identified but there was no significant increase in mRNAs' encoding components of the ubiquitin-proteasome pathway. There was a 6% reduction in polyribosomes (P = 0.007), suggesting a decrease in the capacity for protein synthesis, although there was no measured decrease in the rate of protein synthesis. These findings indicate that high doses of prednisolone lead to a sharp increase in net protein catabolism, which depends more on enhanced protein breakdown, and an uncertain effect on protein synthesis. The mechanisms stimulating proteolysis and the pathway stimulated to increase muscle protein degradation should be explored.


Gating of TonB-dependent transporters by substrate-specific forced remodelling.
Membrane proteins play vital roles in inside-out and outside-in signal transduction by responding to inputs that include mechanical stimuli. Mechanical gating may be mediated by the membrane or by protein(s) but evidence for the latter is scarce. Here we use force spectroscopy, protein engineering and bacterial growth assays to investigate the effects of force on complexes formed between TonB and TonB-dependent transporters (TBDT) from Gram-negative bacteria. We confirm the feasibility of protein-only mediated mechanical gating by demonstrating that the interaction between TonB and BtuB (a TBDT) is sufficiently strong under force to create a channel through the TBDT. In addition, by comparing the dimensions of the force-induced channel in BtuB and a second TBDT (FhuA), we show that the mechanical properties of the interaction are perfectly tuned to their function by inducing formation of a channel whose dimensions are tailored to the ligand.


EMT-related transcription factors and protein stabilization mechanisms involvement in cadherin switch of head and neck squamous cell carcinoma.
Epithelial to mesenchymal transition (EMT) describes a process where epithelial tumor cells acquire mesenchymal characteristics. EMT often correlates with invasion and an increased cell migration potential by losing cellular polarity and cell-cell junctions. It is mainly induced by tumor-microenvironment factors, such as TGF-beta 1 and IL-6, which activate the increased expression of the EMT-transcription factor (TF) Slug. We previously reported the Slug/Krüppel-like factor 4 (KLF4) switch in EMT in HNSCC, and found, that in human papilloma virus (HPV)-negative HNSCC Slug gene expression was significant higher represented, than in HPV-positive HNSCC. The purpose of this study was to investigate the impact of KLF4 and Slug on the regulation of the cadherin switch and on the EMT phenotype. Gene expression of KLF4 positive correlated with E-cadherin in 71 head and neck squamous cell carcinoma (HNSCC) patient tissue samples, which we also confirmed by the investigation of the Cancer Genome Atlas database (TCGA). HPV-transcripts contributed to stabilization of KLF4 at protein level, and simultaneously upregulated E-cadherin. Furthermore, ectopic KLF4 overexpression was associated with epithelial gene expression by induction of E-cadherin, β-catenin and 70-kDa heat shock protein (HSP-70). The presence of HSP-70 ensures the membranous localization of E-cadherin, therefore, the ability of cells to form cadherin/catenin complexes and cellular linkages. In conclusion, KLF4 is a major regulator of the epithelial cadherin-adhesion in HNSCC.


Gene signatures of circulating breast cancer cell models are a source of novel molecular determinants of metastasis and improve circulating tumor cell detection in patients.
Progression to stage IV disease remains the main cause of breast cancer-related deaths. Increasing knowledge on the hematogenous phase of metastasis is key for exploiting the entire window of opportunity to interfere with early dissemination and to achieve a more effective disease control. Recent evidence suggests that circulating tumor cells (CTCs) possess diverse adaptive mechanisms to survive in blood and eventually metastasize, encouraging research into CTC-directed therapies. On the hypothesis that the distinguishing molecular features of CTCs reveal useful information on metastasis biology and disease outcome, we compared the transcriptome of CTCs, primary tumors, lymph-node and lung metastases of the MDA-MB-231 xenograft model, and assessed the biological role of a panel of selected genes, by in vitro and in vivo functional assays, and their clinical significance in M0 and M+ breast cancer patients. We found that hematogenous dissemination is governed by a transcriptional program and identified a CTC signature that includes 192 up-regulated genes, mainly related to cell plasticity and adaptation, and 282 down-regulated genes, involved in chromatin remodeling and transcription. Among genes up-regulated in CTCs, FADS3 was found to increases cell membrane fluidity and promote hematogenous diffusion and lung metastasis formation. TFF3 was observed to be associated with a subset of CTCs with epithelial-like features in the experimental model and in a cohort of 44 breast cancer patients, and to play a role in cell migration, invasion and blood-borne dissemination. The analysis of clinical samples with a panel of CTC-specific genes (ADPRHL1, ELF3, FCF1, TFF1 and TFF3) considerably improved CTC detection as compared with epithelial and tumor-associated markers both in M0 and stage IV patients, and CTC kinetics informed disease relapse in the neoadjuvant setting. Our findings provide evidence on the potential of a CTC-specific molecular profile as source of metastasis-relevant genes in breast cancer experimental models and in patients. Thanks to transcriptome analysis we generated a novel CTC signature in the MDA-MB-231 xenograft model, adding a new piece to the current knowledge on the key players that orchestrate tumor cell hematogenous dissemination and breast cancer metastasis, and expanding the list of CTC-related biomarkers for future validation studies.


Natural killer cells play an important role in virus infection control: Antiviral mechanism, subset expansion and clinical application.
With the global spread of coronavirus disease 2019 (COVID-19), the important role of natural killer (NK) cells in the control of various viral infections attracted more interest, via non-specific activation, such as antibody-dependent cell-mediated cytotoxicity (ADCC) and activating receptors, as well as specific activation, such as memory-like NK generation. In response to different viral infections, NK cells fight viruses in different ways, and different NK subsets proliferate. For instance, cytomegalovirus (CMV) induces NKG2C + CD57 + KIR+ NK cells to expand 3-6 months after hematopoietic stem cell transplantation (HSCT), but human immunodeficiency virus (HIV) induces KIR3DS1+/KIR3DL1 NK cells to expand in the acute phase of infection. However, the similarities and differences among these processes and their molecular mechanisms have not been fully discussed. In this article, we provide a summary and comparison of antiviral mechanisms, unique subset expansion and time periods in peripheral blood and tissues under different conditions of CMV, HIV, Epstein-Barr virus (EBV), COVID-19 and hepatitis B virus (HBV) infections. Accordingly, we also discuss current clinical NK-associated antiviral applications, including cell therapy and NK-related biological agents, and we state the progress and future prospects of NK cell antiviral treatment.


Lu-ECAM-1-mediated adhesion of melanoma cells to endothelium under conditions of flow.
Lu-ECAM-1 is a lung-derived, venular endothelial cell adhesion molecule. It promotes the selective adhesion of lung-metastatic B16-F10 melanoma cells to endothelium under static conditions and mediates colonization of the lungs by the same tumor cells. To test whether Lu-ECAM-1 by itself is sufficient to cause vascular arrest of B16-F10 cells, we measured here under conditions of flow tumor cell adhesion to endothelia that express different amounts of Lu-ECAM-1 on their surfaces. At physiological shear stresses, adhesion of B16-F10 melanoma cells to endothelia correlates positively with the amount of Lu-ECAM-1 expression on the endothelial cell surface and inversely with the level of the applied shear stress. Tumor cell trajectories are biphasic; i.e., B16-F10 melanoma cells initially move along the endothelial surface with a velocity similar to the theoretical velocity, then arrest within a fraction of a second. Arrest is permanent for most B16-F10 melanoma cells at all shear stresses tested. Tumor cells never engaged in a rolling motion prior to arrest. Masking of the Lu-ECAM-1 ligand on the surface of B16-F10 melanoma cells with soluble Lu-ECAM-1 impedes arrest of tumor cells on the surface of the test endothelium. Purified Lu-ECAM-1 also mediates B16-F10 arrest, but arrest is mostly transient at shear stresses of 0.59 dynes/cm2 and higher, implying adhesion by single receptor/ligand bonds. Our data suggest that Lu-ECAM-1 plays a critical role in the recognition and initial arrest of murine melanoma cells in lung venules.


Effects of metabolic memory on inflammation and fibrosis associated with diabetic kidney disease: an epigenetic perspective.
Diabetic kidney disease (DKD) is one of the most common microvascular complication of both type 1 (T1DM) and type 2 diabetes mellitus (T2DM), and the leading cause of end-stage renal disease (ESRD) worldwide. Persistent inflammation and subsequent chronic fibrosis are major causes of loss of renal function, which is associated with the progression of DKD to ESRD. In fact, DKD progression is affected by a combination of genetic and environmental factors. Approximately, one-third of diabetic patients progress to develop DKD despite intensive glycemic control, which propose an essential concept "metabolic memory." Epigenetic modifications, an extensively studied mechanism of metabolic memory, have been shown to contribute to the susceptibility to develop DKD. Epigenetic modifications also play a regulatory role in the interactions between the genes and the environmental factors. The epigenetic contributions to the processes of inflammation and fibrogenesis involved in DKD occur at different regulatory levels, including DNA methylation, histone modification and non-coding RNA modulation. Compared with genetic factors, epigenetics represents a new therapeutic frontier in understanding the development DKD and may lead to therapeutic breakthroughs due to the possibility to reverse these modifications therapeutically. Early recognition of epigenetic events and biomarkers is crucial for timely diagnosis and intervention of DKD, and for the prevention of the progression of DKD to ESRD. Herein, we will review the latest epigenetic mechanisms involved in the renal pathology of both type 1 (T1DN) and type 2 diabetic nephropathy (T2DN) and highlight the emerging role and possible therapeutic strategies based on the understanding of the role of epigenetics in DKD-associated inflammation and fibrogenesis.


Cyber-Attacks Risk Analysis Method for Different Levels of Automation of Mining Processes in Mines Based on Fuzzy Theory Use.
The rising automation level and development of the Industry 4.0 concept in the mining sector increase the risk of cyber-attacks. As a result, this article focuses on developing a risk analysis method that integrates Kaplan's and Garrick's approach and fuzzy theory. The proposed approach takes into account the level of automation of the operating mining processes. Moreover, it follows five main steps, including identifying the automation level in a selected mine, definition of cyber-attack targets, identification of cyber-attack techniques, definition of cyber-attack consequences, and risk ratio assessment. The proposed risk assessment procedure was performed according to three cyber-attack targets (databases, internal networks, machinery) and seven selected types of cyber-attack techniques. The fuzzy theory is implemented in risk parameter estimation for cyber-attack scenario occurrence in the mining industry. To illustrate the given method's applicability, seven scenarios for three levels of mine automation are analyzed. The proposed method may be used to reveal the current cybersecurity status of the mine. Moreover, it will be a valuable guide for mines in which automation is planned in the near future.


Insecticide resistance status of Aedes aegypti in southern and northern Ghana.
Outbreaks of Aedes-borne arboviral diseases are becoming rampant in Africa. In Ghana, there is no organized arboviral control programme with interventions restricted to mitigate outbreaks. Insecticide application is a crucial part of outbreak responses and future preventative control measures. Thus, knowledge of the resistance status and underlying mechanisms of Aedes populations is required to ensure optimal insecticide choices. The present study assessed the insecticide resistance status of Aedes aegypti populations from southern Ghana (Accra, Tema and Ada Foah) and northern Ghana (Navrongo) respectively. Phenotypic resistance was determined with WHO susceptibility tests using Ae. aegypti collected as larvae and reared into adults. Knockdown resistance (kdr) mutations were detected using allele-specific PCR. Synergist assays were performed with piperonyl butoxide (PBO) to investigate the possible involvement of metabolic mechanisms in resistance phenotypes. Resistance to DDT was moderate to high across sites (11.3 to 75.8%) and, for the pyrethroids deltamethrin and permethrin, moderate resistance was detected (62.5 to 88.8%). The 1534C kdr and 1016I kdr alleles were common in all sites (0.65 to 1) and may be on a trajectory toward fixation. In addition, a third kdr mutant, V410L, was detected at lower frequencies (0.03 to 0.31). Pre-exposure to PBO significantly increased the susceptibility of Ae. aegypti to deltamethrin and permethrin (P < 0.001). This indicates that in addition to kdr mutants, metabolic enzymes (monooxygenases) may be involved in the resistance phenotypes observed in the Ae. aegypti populations in these sites. Insecticide resistance underpinned by multiple mechanisms in Ae. aegypti indicates the need for surveillance to assist in developing appropriate vector control strategies for arboviral disease control in Ghana.


Reconstitution of mouse spermatogonial stem cell niches in culture.
Spermatogonial stem cells (SSCs) reside in specific niches within seminiferous tubules. These niches are thought to secrete chemotactic factors for SSCs, because SSCs migrate to them upon transplantation. However, the identity of these chemotactic molecules remains unknown. Here, we established a testis feeder cell culture system and used it to identify SSC chemotactic factors. When seeded on testis cells from infertile mice, SSCs migrated beneath the Sertoli cells and formed colonies with a cobblestone appearance that were very similar to those produced by hematopoietic stem cells. Cultured cells maintained SSC activity and fertility for at least 5 months. Cobblestone colony formation depended on GDNF and CXCL12, and dominant-negative GDNF receptor transfection or CXCL12 receptor deficiency reduced SSC colonization. Moreover, GDNF upregulated CXCL12 receptor expression, and CXCL12 transfection in Sertoli cells increased homing efficiency. Overall, our findings identify GDNF and CXCL12 as SSC chemotactic factors in vitro and in vivo.


Enhancement of the photostability and photoactivity of metallo-meso-5,10,15,20-tetrakis-(4-sulfonatophenyl)porphyrins by polymeric micelles.
Metallo-meso-5,10,15,20-tetrakis-(4-sulfonatophenyl)porphyrins (metallo-TPPSs), such as ZnTPPS, have been widely used as photosensitizers. However, their vulnerability to photodegradation significantly limits their applications. In this contribution, we demonstrate a method to enhance the photostability of metallo-TPPSs while retaining photoactivity via encapsulation inside cores of complex micelles. Poly(ethylene glycol)-b-poly(4-vinylpyridine) (PEG-b-P4VP) and metallo-TPPSs can form complex micelles in acidic solution through electrostatic interactions and then undergo axial coordination with the pyridine moieties of PEG-b-P4VP when the pH is adjusted to 7.4. In this way, metallo-TPPSs are entrapped in the hydrophobic, compact micellar cores, which effectively prevents photodegradation of the metallo-TPPSs that would otherwise occur in aqueous solution. In addition, the photodebromination of 2,3-dibromo-3-phenylpropionic acid (DPP) sensitized with ZnTPPS has been used as a model reaction to study the photosensitive activity of ZnTPPS entrapped in complex micelles. The entrapped ZnTPPSs exhibit pronounced activity and have much higher efficiency and faster photosensitive reaction rates than free ZnTPPS.


X-ray fluorescence analysis of lead in teeth of urban children in situ: correlation between the tooth lead level and the concentration of blood lead and free erythroporphyrins.
The tooth lead level of 30 lead-exposed children was measured in situ using an X-ray fluorescence technique. The tooth lead concentration divided by the child's age correlated with the free erythroporphyrin (FEP) (r = 0.51) and the blood lead levels (r = 0.31). The mean tooth lead concentrations of 10 Class IV children (FEP greater than or equal to 190 micrograms/100 ml; blood lead 30-80 micrograms/ml) was 14.5 +/- 5.5 ppm/year. Reexamination of the histories of children who had been classified as Class II or III, but who had tooth lead concentrations within 1 SD of the Class IV level, indicated that many of these children could also be considered to be Class IV children. When the x-ray fluorescence technique was used to screen an urban population of 300 children, the tooth lead values indicated that 72% of the children had been exposed to low levels of environmental lead. Six percent of the children were found to have tooth lead concentrations in excess of 9 ppm/year and within 1 SD of the mean value exhibited by Class IV children. These children were considered to have a high body lead burden. The percentage of children having an elevated tooth lead level is similar to the number previously reported using exfoliated deciduous teeth.


Genome profiling of uropathogenic E. coli from strictly defined community-acquired UTI in paediatric patients: a multicentric study.
Urinary tract infection (UTI) in children is a common bacterial infection. The emergence of extended-spectrum beta-lactamases (ESBLs) poses a major challenge against the treatment of uropathogens. We aimed to characterize the E. coli isolates recovered from children with UTI for their resistance profile and circulating sequence types (ST). Children (> 1.5-18 years of age) from different community health centres of India with symptoms of UTI were enrolled. Isolates causing significant bacteriuria were identified by Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) and tested for antimicrobial susceptibility by the automated system, VITEK-2 (Biomeriux, Durhum, US). Nineteen E. coli isolates (15 ESBL positive and 4 ESBL negative) were sequenced in Oxford Nanopore platform followed by core-genome phylogeny, accessory genome cluster analysis, identification of sequence types, mobile genetic elements, genetic antimicrobial resistance markers. The correlation between detection of antimicrobial resistance genes with phenotypic resistance profiles was also investigated. Eleven percent of children had significant bacteriuria [male:female-1:1, > 50% were 11-18 years of age group]. E. coli was predominant (86%) followed by K. pneumoniae (11%). Susceptibility of E. coli was highest against fosfomycin (100%) followed by carbapenems (90.7%) and nitrofurantoin (88.8%). ST131 (15.8%) and ST167 (10.5%) found as high-risk clones with the presence of plasmid [IncFIB (63.1%), IncFIA (52.6%)], and composite transposon [Tn2680 (46.6%)] in many isolates. Few isolates coharboured multiple beta-lactamases including blaNDM-5 (33.3%), blaOXA-1 (53.3%), blaCTX-M-15 (60%) and blaTEM-4 (60%). This study highlights horizontal transmission of resistance genes and plasmids in paediatric patients at community centers across the nation harbouring multidrug-resistant genes such as blaNDM-5 and blaCTX-M-15 associated with high-risk clones ST131 and ST167. The data is alarming and emphasizes the need for rapid identification of resistance markers to reduce the spread in community. To our knowledge, this is the first multicentric study targeting paediatric UTI patients from the community setting of India.


CORRELATION BETWEEN FOCAL CHOROIDAL EXCAVATION AND UNDERLYING RETINOCHOROIDAL DISEASE: A Pathological Hypothesis From Clinical Observation.
To evaluate focal choroidal excavation (FCE) in eyes with various diseases using multimodal imaging modalities and to investigate the correlation of FCE and underlying chorioretinal diseases. This retrospective observational study included 62 eyes from 56 patients who were identified by optical coherence tomography as having FCE. All included patients underwent comprehensive clinical examinations and multimodal imaging to identify and detect the characteristics of FCE and its correlation with underlying chorioretinal diseases. All included patients were of Chinese descent, and the median age at diagnosis was 43 years (range: 15-66). Seventy-three FCEs appeared in these included eyes. Most FCEs were formed at sites with anatomical changes caused by various chorioretinal diseases. Choroidal osteoma, punctate inner choroidopathy, and central serous chorioretinopathy were the most common etiologies of FCE. During follow-up, 14 eyes (22.58%) exhibited a pattern change and three eyes (4.84%) developed new-onset choroid neovascularization. Focal choroidal excavation is a common sign found in a variety of chorioretinal diseases. Processes that involve impairment or tissue loss of the outer retina and inner choroid and disrupt the balance of intraocular pressure and choroidal pressure because of mechanical disturbance may play a role in FCE formation.


Attitudes to AIDS: a comparative analysis of a new and negative stereotype.
Two propositions about attitudes, which have previously been supported with respect to the mentally ill, were examined with respect to AIDS patients. The first, that people attach a stigma to the AIDS patient, was strongly supported, and two quite independent components of the stigma were found. One of these components identified as dependence, was closely related to the attributes of typical cancer patients and coronary heart patients, while the other, identified as low moral worth, clearly distinguished the AIDS patient from the other two groups of patients. The second proposition, that attitudes to AIDS are not strongly related to age, sex and occupational background, was largely supported. However there was some evidence that males rated AIDS patients lower on moral worth than did females.


Abdominal pregnancy diagnosed by ultrasonography and treated successfully by laparotomy: Two cases report.
Abdominal pregnancy is one of the rare types of ectopic pregnancies that has a high rate of mortality. Its clinical presentation is similar to those of the other types of ectopic pregnancies. we report two cases of abdominal pregnancies whose the diagnosis was made by ultra-sonography and successfully treated by laparotomy in emergency obstetrical department of Ibn Rochd University Hospital of Casablanca. CLINICAL DISCUSSION: the ultra-sonography based on precise criteria represents the fundamental paraclinical examination in the diagnosis of abdominal pregnancies. The treatment of early form is based on surgery; And despite the advent of laparoscopic surgery, laparotomy retains its indications including forms with hemorrhagic shock. Due to the high risk of complications and maternal mortality, abdominal pregnancies should be surgically treated as soon as possible when the diagnosis is confirmed.


A deep attention model to forecast the Length Of Stay and the in-hospital mortality right on admission from ICD codes and demographic data.
Leveraging the Electronic Health Records (EHR) longitudinal data to produce actionable clinical insights has always been a critical issue for recent studies. Non-forecasted extended hospitalizations account for a disproportionate amount of resource use, the mediocre quality of inpatient care, and avoidable fatalities. The capability to predict the Length of Stay (LoS) and mortality in the early stages of the admission provides opportunities to improve care and prevent many preventable losses. Forecasting the in-hospital mortality is important in providing clinicians with enough insights to make decisions and hospitals to allocate resources, hence predicting the LoS and mortality within the first day of admission is a difficult but a paramount endeavor. The biggest challenge is that few data are available by this time, thus the prediction has to bring in the previous admissions history and free text diagnosis that are recorded immediately on admission. We propose a model that uses the multi-modal EHR structured medical codes and key demographic information to classify the LoS in 3 classes; Short Los (LoS⩽10 days), Medium LoS (10<LoS⩽30 days) and Long LoS (LoS>30 days) as well as mortality as a binary classification of a patient's death during current admission. The prediction has to use data available only within 24 h of admission. The key predictors include previous ICD9 diagnosis codes, ICD9 procedures, key demographic data, and free text diagnosis of the current admission recorded right on admission. We propose a Hierarchical Attention Network (HAN-LoS and HAN-Mor) model and train it to a dataset of over 45321 admissions recorded in the de-identified MIMIC-III dataset. For improved prediction, our attention mechanisms can focus on the most influential past admissions and most influential codes in these admissions. For fair performance evaluation, we implemented and compared the HAN model with previous approaches. With dataset balancing techniques HAN-LoS achieved an AUROC of over 0.82 and a Micro-F1 score of 0.24 and HAN-Mor achieved AUC-ROC of 0.87 hence outperforming the existing baselines that use structured medical codes as well as clinical time series for LoS and Mortality forecasting. By predicting mortality and LoS using the same model, we show that with little tuning the proposed model can be used for other clinical predictive tasks like phenotyping, decompensation,re-admission prediction, and survival analysis.


Factors influencing left ventricular thrombus resolution and its significance on clinical outcomes.
A left ventricular thrombus (LVT) is not uncommon in patients with impaired LV systolic function. However, the treatment strategy for LVT has not yet been fully established. We aimed to identify the factors influencing LVT resolution and the significance of LVT resolution on clinical outcomes. We retrospectively investigated patients diagnosed with LVT with left ventricular ejection fraction (LVEF) < 50% on transthoracic echocardiography from January 2010 to July 2021 in a single tertiary centre. LVT resolution was monitored through serial follow-up transthoracic echocardiography. The primary clinical outcome was a composite of all-cause death, stroke, transient ischaemic attack, and arterial thromboembolic events. LVT recurrence was also evaluated in patients with LVT resolution. There were 212 patients diagnosed with LVT (mean age, 60.5 ± 14.0 years; male, 82.5%). The mean LVEF was 33.1 ± 10.9%, and 71.7% of patients were diagnosed with ischaemic cardiomyopathy. Most patients were treated with vitamin K antagonists (86.7%), and 28 patients (13.2%) were treated with direct oral anticoagulants or low molecular weight heparin. LVT resolution was observed in 179 patients (84.4%). LVEF improvement failure within 6 months was a significant factor hindering LVT resolution (hazard ratio, HR: 0.52, 95% confidence interval, CI: 0.31-0.85, P = 0.010). During a median 4.0 years of follow-up (interquartile range, IQR: 1.9 to 7.3 years), 32 patients (15.1%) experienced primary outcomes (18 all-cause deaths, 15 strokes, and 3 arterial thromboembolisms) and 20 patients (11.2%) experienced LVT recurrence after LVT resolution. LVT resolution was independently associated with a lower risk for primary outcomes (HR: 0.45, 95% CI: 0.21-0.98, P = 0.045). In the patients with resolved LVT, discontinuation or duration of anticoagulation after resolution were not significant predictors for LVT recurrence, but LVEF improvement failure at LVT resolution was associated with a significantly higher risk of LVT recurrence (HR: 3.10, 95% CI: 1.23-7.78, P = 0.016). This study suggests that LVT resolution is an important predictor for favourable clinical outcomes. LVEF improvement failure interfered with LVT resolution and appeared to be a crucial factor for LVT recurrence. After LVT resolution, continuation of anticoagulation did not seem to impact LVT recurrence and the prognosis.


The Malay-Version Knowledge, Risk Perception, Attitude and Practice Questionnaire on Heatwaves: Development and Construct Validation.
Heatwaves have long been recognised as a serious public health concern. This study was aimed at developing and validating a Malay-version of a questionnaire for evaluating knowledge, risk perception, attitudes, and practices regarding heatwaves. The knowledge construct was evaluated with item analysis and internal reliability. The psychometric characteristics, construct and discriminant validity, and internal consistency of the risk perception, attitude and practice constructs were evaluated with exploratory factor analysis (EFA) and confirmatory factor analysis (CFA). The 16 items in the knowledge construct had a good difficulty, discrimination, and reliability index of 0.81. A total of 16 items were maintained in EFA with Cronbach's alpha of 0.84 and 0.82, 0.78 and 0.84 obtained for total items and risk perception, attitude, and practice constructs, respectively. A total of 15 items were retained after CFA. The finalised model met the fitness indices threshold. The convergent and discriminant validity were good. This newly developed Malay-version KRPAP questionnaire is reliable and valid for assessing Malaysians' knowledge, risk perception, attitudes, and practices regarding heatwaves.


Peripheral neuromodulation in women with overactive bladder syndrome: a systematic review.
Overactive bladder syndrome (OBS) is a urological disorder characterized by urinary urgency, increased frequency, nocturia, and may be associated with urge urinary incontinence. Posterior tibial nerve peripheral neuromodulation (PTNS) is globally recognized within the treatment options available, although an optimal protocol has not been tested. The objective of this review is to collect the evidence available on the most widely used protocol of the PTNS technique in obtaining good results in the treatment for women with OBS. A systematic review of the scientific literature was carried out in PubMed, Embase, WoS and Scopus databases. A total of 222 results were obtained, with 58 duplicates, of which 8 studies met the established inclusion criteria, all of them randomized clinical trials. The included works show that the most widely used PTNS technique with beneficial results in women with OBS is the percutaneous route with a needle placed 5-6cm proximal to the tibial malleolus, posterior to the edge of the tibia, with the following parameters: 1 weekly session of 30minutes duration for 12 weeks, the use of pulse widths of 200μs with frequency of 20Hz. PTNS may have beneficial and safe short-term effects in women with OBS. Despite showing statistically significant improvements in clinical symptoms, further research is needed to obtain clear scientific evidence on the optimal protocol for treating women with OBS.


Primitive streak-forming cells of the chick invaginate through a basement membrane.
Recently fibronectin was shown to appear in the development of the chick for the first time as a thin band on the epiblastic side facing the hypoblast just prior to primitive streak formation. It was thus suggested that fibronectin might be instrumental in the migration of cells that lead to axis formation during primitive streak formation. In the present work we have examined simultaneously for the presence of fibronectin and the specific basement membrane glycoprotein laminin during primitive streak formation using immunofluorescence methods. Laminin was found to be expressed between the epiblast and the hypoblast of stage XIII1 chick blastoderms. During the immediately following process of streak formation the laminin was found to be continuously detectable throughout the area covered by the hypoblast, but disrupted on the streak area. Fibronectin was found to co-distribute with laminin in stage XIII and in the early primitive streak chick blastoderms. It is concluded that at stage XIII laminin and fibronectin form part of a basement membrane that is partially disrupted during the immediately following process of primitive streak formation in order to allow the migration of the streak-forming epiblastic cells during this morphogenetic process.


[A Case of Stenosis at the Junction of the Renal Pelvis and Ureter after Laparoscopic Right Hemicolectomy].
A 50s year old woman was referred to former hospital for abdominal pain and melena. Colonoscopy revealed an ascending colon cancer(cT4aN1bM0, cStage Ⅲb, UICC)and gastrointestinal endoscopy revealed an esophagogastric junction cancer(cT1bN0M0, cStage ⅠA, UICC). Preoperative CT showed mild bilateral hydronephrosis. Laparoscopic right hemicolectomy( Lap-RHC, D3)was performed for ascending colon cancer then, and abdominal CT was taken 2 days before the robot-assisted proximal gastrectomy(RAPG, D1+). Right hydronephrosis was observed. No evidence of obvious ureteral damage was found during Lap-RHC. When retrograde pelvic ureterography was performed and a stenosis was found at the right renal pelvis and ureteral junction. It is considered that the ureter stenosis became prominent due to the postoperative inflammatory change of Lap-RHC. RAPG was performed as scheduled. Two months after discharge, a renal fistula was constructed. Pyeloplasty is scheduled to be performed in the future.


Nuclear lipid-dependent signal transduction in human osteosarcoma cells.
The enzymes and substrates involved in phosphoinositide signal transduction which have been detected in the nucleus of several cell types have been demonstrated to be responsive to agonists. The complexity of this aspect of inositide function has been previously analyzed in some cell models characterized by a mitogenic or differentiating response to specific factors. An interesting experimental model is represented by human derived osteosarcoma Saos-2 cells, characterized by the expression of high affinity receptors for interleukin 1 alpha (IL-1 alpha), which is one of the most potent stimulators of bone resorption. In particular, we investigated the earliest intracellular events following the binding of IL-1 alpha to its receptor, involving the inositide signal transduction pathway. Saos-2 cells present a partitioning of the phosphoinositidase (PLC) isoforms; in fact, the nucleus contains both PLC beta 1 and gamma 1, while the cytoplasm contains almost exclusively the gamma 1 isoform. IL-1 alpha evokes a rapid and transient increase of the PLC beta 1 activity in the nucleus, which causes the hydrolysis of phosphatidylinositol mono- and bis-phosphate. In response to IL-1 alpha, not only the canonical inositol lipid pathway appears to be involved; also the 3'-phosphorylated lipids generated by phosphatidylinositol 3-kinase (PI 3-K), which may act as second messengers, appear to be affected. In fact, Saos-2 cells present a nuclear PI 3-K activity which can be enhanced by the IL-1 alpha treatment. Among the possible targets of the second messengers released by the nuclear PLC beta 1 activation, we found that some protein kinase C isoforms, namely the epsilon and zeta, which are present within the nucleus, are activated after IL-1 alpha exposure. These activated PKC isoforms, in turn, could modulate the activity of the transcription factor NFkB, which, 5 min after IL-1 alpha treatment, has already translocated to the nucleus and bound to DNA to promote gene activation. The actual role of the inositide pathway in the Saos-2 cell function has also been investigated by utilizing cell clones transfected with the mouse sequence of the PLC beta 1.


Production of an antigenic peptide by insulin-degrading enzyme.
Most antigenic peptides presented by major histocompatibility complex (MHC) class I molecules are produced by the proteasome. Here we show that a proteasome-independent peptide derived from the human tumor protein MAGE-A3 is produced directly by insulin-degrading enzyme (IDE), a cytosolic metallopeptidase. Cytotoxic T lymphocyte recognition of tumor cells was reduced after metallopeptidase inhibition or IDE silencing. Separate inhibition of the metallopeptidase and the proteasome impaired degradation of MAGE-A3 proteins, and simultaneous inhibition of both further stabilized MAGE-A3 proteins. These results suggest that MAGE-A3 proteins are degraded along two parallel pathways that involve either the proteasome or IDE and produce different sets of antigenic peptides presented by MHC class I molecules.


CYP19 gene expression in subcutaneous adipose tissue is associated with blood pressure in women with polycystic ovary syndrome.
In polycystic ovary syndrome (PCOS), hypertension has been linked to androgen excess and insulin resistance. Aromatase, an enzyme encoded by the CYP19 gene, affects androgen metabolism and estrogen synthesis, influencing the androgen to estrogen balance. We characterized CYP19 gene expression in subcutaneous adipose tissue of women with PCOS and normal controls and evaluated the association between subcutaneous fat CYP19 mRNA, circulating hormone levels, and blood pressure. This case-control study was carried out with 31 PCOS patients and 27 BMI-matched normotensive non-hirsute women with regular cycles. Participants underwent anthropometric measurements, collection of blood samples, and adipose tissue biopsy (28 PCOS and 19 controls). Hypertension was defined as systolic blood pressure ≥ 130 mmHg and/or diastolic blood pressure ≥ 85 mmHg. PCOS patients were divided into normotensive and hypertensive. Main outcome measures were serum estrogen and androgen levels, estrogen-to-androgen ratio, and CYP19 gene expression in subcutaneous fat. Subcutaneous CYP19 mRNA was higher in hypertensive PCOS than in control and normotensive PCOS women (p = 0.014). Estrogen-to-androgen ratio was lower in hypertensive PCOS than controls (p < 0.003). Estrogen-to-androgen ratio ≤ 0.06 (median for the three groups) was observed in 91% of hypertensive PCOS women, vs. 37% and 61% in the control and normotensive PCOS groups (p = 0.011). CYP19 gene expression in subcutaneous fat of PCOS patient correlated positively with systolic (p = 0.006) and diastolic blood pressure (p = 0.009). Androgen excess and hyperinsulinemia may play a role in the molecular mechanisms that activate aromatase mRNA transcription in abdominal fat tissue.


Transfer RNA control of the activation of isomeric tRNATrp's.
Previous studies of the homologous aminoacylations of Escherichia coli and yeast tRNATrp's terminating in 2'- and 3'-deoxyadenosine established that E. coli tryptophanyl-tRNA synthetase activates its cognate tRNA preferentially on the 2' position, while the corresponding yeast enzyme utilizes the 3' position on its homologous substrate tRNA. As this seemed to be the only change in positional specificity during evolution, the heterologous activations were investigated in an effort to determine the basis for this change. Remarkably, E. coli tRNATrp terminating in 3'-deoxyadenosine was found to be the preferred substrate for both the E. coli and yeast activating enzymes, while the same tryptophanyl-tRNA synthetase preparations both activated the isomeric yeast tRNATrp's preferentially on the 3' position. Thus, the preferred position of activation was found to be specified by the tRNA rather than the activating enzyme and, additionally, to be due to some process not reflected in initial velocity measurements. The variable utilization of individual modified aminoacyl-tRNA's as substrates in an enzyme-catalyzed deacylation process appears to provide the most likely explanation for the experimental observations.


Normal whole blood Bohr effect in Peruvian natives of high altitude.
Measurements of whole blood O2 affinity were made with an automatic technique that allowed recording of O2 saturation continuously over a range of PO2 of 1 to 150 torr at constant pH and PCO2 with fresh samples of blood obtained from 5 natives of high altitude at Morococha, Peru (altitude, 4540 m). The results were not significantly different from those obtained with controls living at sea level with regard to the dlog P50/dpH slope at constant PCO2 or at constant base excess. Both results also agree with previous studies with normal sea-level blood. Thus earlier claims of an increased Bohr in Peruvian high-altitude natives is not supported.


Correlation of Severity of COVID-19 Disease With Gastrointestinal Manifestations and Liver Injury - A North Brooklyn Community Hospital Experience: A Retrospective Cohort Study.
Introduction The primary receptor for SARS-CoV-2 infection, angiotensin-converting enzyme-2 (ACE-2), is expressed in the gastrointestinal tract and liver parenchyma. The involvement of the gastrointestinal tract with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection has remained unclear. The following study retrospectively reviews gastrointestinal symptoms and liver function tests at the time of hospital admission to identify patient outcomes including prolonged hospital stay, the requirement for intensive care, and all-cause in-hospital 30-day mortality. Methods A retrospective review of patient charts at the Woodhull Medical and Mental Health Center (WMC) was conducted at the time of hospital admission, using a pre-determined selection criterion. All adult patients, both inpatient and outpatient, were included from March 2020 till May 2020. A 95% confidence interval was used to estimate the odds ratio (OR) for patient outcomes. Results Of the 520 patients, gastrointestinal symptoms including nausea (OR = 0.375, p = 0.015), and nausea and vomiting in combination (OR = 0.400, p = 0.016) had an inverse protective relationship with all-cause in-hospital 30-day mortality among COVID-19 patients. Gastrointestinal symptoms including diarrhea (OR = 1.008, p < 0.001), and nausea and vomiting (OR = 1.291, p = 0.043) had a mild impact on the length of hospital stay. Conclusion Elevated liver transaminases including alanine transaminase (ALT) and aspartate transaminase (AST) at the time of hospital admission can predict critical care requirement and all-cause 30-day hospital mortality in patients with COVID-19 infection. Presence of gastrointestinal symptoms is associated with worsened outcomes.


Serum paraoxonase-I activity is unaffected by short-term administration of simvastatin, bezafibrate, and their combination in type 2 diabetes mellitus.
The high-density lipoprotein (HDL)-associated anti-oxidative and anti-inflammatory enzyme, paraoxonase-I, has been found previously to be lower in type 2 diabetes mellitus. We studied whether statin and fibrate treatment, alone and in combination, affect serum paraoxonase-I activity in conjunction with changes in HDL cholesterol in diabetic patients. A placebo-controlled crossover study was carried out in 14 type 2 diabetic patients to test the effect of 8 weeks of active treatment with simvastatin (40 mg daily), bezafibrate (400 mg daily), and their combination on serum paraoxonase-I activity, measured as its activity towards arylesterase and paraoxon. Serum paraoxonase-I activity was also compared between these diabetic patients and 49 non-diabetic control subjects. Serum arylesterase activity was lower in type 2 diabetic patients compared to control subjects (P < 0.001), but the difference in paraoxonase activity was not significant (P = 0.22). Neither arylesterase (P = 0.24) nor paraoxonase activity (P = 0.37) was increased in response to treatment, despite higher HDL cholesterol and apolipoprotein A-I during combination therapy (P < 0.05 for both). Short-term administration of simvastatin and bezafibrate, even when combined, is ineffective in raising serum paraoxonase-I activity in type 2 diabetes.


The influence of living conditions and individual behaviors on the oral-systemic disease connection: a cross-sectional analysis.
To determine the extent to which living conditions and individual behaviors influence the association between oral health status and systemic disease outcomes in Ontario, Canada's most populated province. A secondary data analysis of Ontario data from the Canadian Community. Health Survey 2013/14 was undertaken. Separate analyses were conducted for participants aged 35-59 years (n = 11,858) and 60+ years (n = 11,273). A series of regression models were constructed to examine the association between self-reported oral health status and systemic disease outcomes (arthritis, diabetes, hypertension, heart disease, chronic obstructive pulmonary disease, and stroke). Models were adjusted by proxies of living conditions (income, education, ethnicity, country of birth, employment, and food security) and individual behaviors (smoking status, alcohol use, tooth brushing, life stress, physical activity, sense of belonging). Percent attenuation between models was calculated to determine the extent of the living condition-behavior impact. In both age groups, the prevalence of arthritis and high blood pressure was the highest, followed by heart disease. There was variation in percent attenuation by age group and outcome. Among participants aged 35-59 years, living conditions had a greater impact on the oral-systemic relationship, while individual behaviors played a greater role in this association among adults aged 60+ years. There is an association between oral and systemic diseases; however, after accounting for living conditions and individual behaviors, this relationship was attenuated. This highlights the need to address upstream and midstream factors that are common to oral and systemic conditions.


High frequency of class 3 allele in the human insulin gene in Japanese type 2 (non-insulin-dependent) diabetic patients with a family history of diabetes.
The restriction fragment length polymorphism in the 5' flanking region of the human insulin gene was studied in 155 nonobese Japanese subjects. The subjects consisted of 36 Type 2 (non-insulin-dependent) diabetic patients with a family history of diabetes mellitus, 42 Type 2 diabetic patients without a family history of diabetes, 42 Type 1 (insulin-dependent) diabetic patients, and 35 healthy volunteers who served as control subjects. It was demonstrated that, in Japanese healthy subjects and diabetic patients, the incidence of the insertion into 5' flanking region of the insulin gene was found to be significantly lower (p less than 0.05) than those in Caucasians and other races already investigated. Even though the class 3 gene allelic frequency in Type 2 diabetic patients without a family history of diabetes (0.060) was not higher than that in healthy subjects (0.014), in nonobese Type 2 diabetic patients with a family history of diabetes the allelic frequency of the inserted class 3 gene (0.111) was found to be significantly higher (p less than 0.02) than that in control subjects. These data suggest that the insulin gene polymorphism relates to the aetiology of diabetes mellitus.


Involvement of reactive oxygen species in post-ischaemic flap necrosis and its prevention by antioxidants.
In attempts to substantiate the possible participation of reactive oxygen species, and the significance of the xanthine oxidase system in both post-ischaemic reperfusion necrosis of the island flap and distal necrosis of the pedicle flap, and to develop new pharmacological measures for salvaging flap necrosis, a series of experiments were made using an island flap model and a random-pattern flap model in rats. The results were as follows: (1) Epoxysuccinyl derivative (E-64c), allopurinol and L-SOD salvaged post-ischaemic reperfusion necrosis of the island flaps; (2) E-64c and allopurinol did not salvage anticipated necrosis of the distal region of random flaps but L-SOD did; (3) tissue SOD activity did not reflect the fate of the island flap, but did of the distal region of the random flap. These results demonstrated a possible involvement of ROS in both post-ischaemic necrosis of island flaps and distal necrosis of random flaps. However, xanthine oxidase was significant in producing ROS only in the former.


CD8+ T Cell Responses to Toxoplasma gondii: Lessons from a Successful Parasite.
Toxoplasma gondii infection in mice provides an excellent model for the study of CD8+ T cell responses. Natural and engineered T. gondii antigens have led the way to understanding the factors regulating antigen presentation from vacuolar pathogens. T. gondii infection of resistant and sensitive mouse strains provides unique models to study both effective CD8+ T cell function and protection in a well-controlled infection attributed to a novel T cell population, and T cell exhaustion in a progressing chronic infection. Additionally, the long-term persistence of the parasite in the brain provides a unique model of neurotropic infection used to study CD8+ T cell entry, retention, and function in the brain. Here we discuss recent advances in each of these areas.


Novel fluorogenic calix[4]arene-bis(crown-6-ether) for selective recognition of thallium(I).
A new fluorogenic, dansyl group-containing derivative of 1,3-alternate calix[4]arene-bis(crown-6-ether) provides optical recognition of Tl+ with selectivity over many other metal cations, including Na+, K+, Ca2+, Ag+, Hg2+ and Pb2+, and embodies the first example of a calixarene-based fluorescent Tl+-chemosensor.


Application of a new computer-assisted robotic visualization system in cochlear implantation-Proof of concept.
Over the last decades conventional cochlear implant (CI) surgery has remained essentially unchanged. Nevertheless, alternative implantation techniques to further improve patient outcomes such as endaural implantation or robot-assisted surgery have been proposed in recent years. However, none of these have gained acceptance in clinical routine, thus confirming a demand for new developments. Cochlear implant surgery was performed in two mastoid bones obtained from body donors using a novel hands-free exoscope. Advantages and disadvantages of the system were evaluated. In all cases, implantation of the electrode was feasible. The system allowed for hands-free movement and adjustment of the exoscope by the head-mounted display. Network connectivity of the system leaves room for improvement. The RoboticScope is an innovative tool and can be used supportively in conventional CI surgery in the experimental setting. Although operating the device requires a certain learning curve, the usability is intuitive for every ear surgeon.


The relation of chlamydial infection of the cervix to time elapsed from the onset of menses.
To investigate the relation of chlamydial infection of the cervix to time elapsed from the onset of menses, we prospectively studied 338 women receiving routine gynecologic care in two clinical sites in which the prevalence of chlamydial infection was 3 and 6%. Eleven (13%) of 86 women evaluated in the fourth or fifth week after the onset of menses (days 22-35) had chlamydial infection compared to 4 (1.6%) of 252 women seen in the first 3 weeks (days 1-21) (p less than 0.001). For women seen in the fourth or fifth week, the relative risk of chlamydial infection was 8.1 (95% CI, 3.1-20.6). Controlling for known risk factors for chlamydial infection with multiple logistic regression analysis, the relative risk was sustained (RR 10.8; 95% CI, 3.1-37.4). We conclude that time elapsed from the onset of menses is an important independent risk factor for chlamydial infection of the cervix that may lead to more effective and efficient screening strategies.


Metabolic Profiling and Compound-Class Identification Reveal Alterations in Serum Triglyceride Levels in Mice Immunized with Human Vaccine Adjuvant Alum.
Alum has been widely used as an adjuvant for human vaccines; however, the impact of Alum on host metabolism remains largely unknown. Herein, we applied mass spectrometry (MS) (liquid chromatography-MS)-based metabolic and lipid profiling to monitor the effects of the Alum adjuvant on mouse serum at 6, 24, 72, and 168 h post-vaccination. We propose a new strategy termed subclass identification and annotation for metabolomics for class-wise identification of untargeted metabolomics data generated from high-resolution MS. Using this approach, we identified and validated the levels of several lipids in mouse serum that were significantly altered following Alum administration. These lipids showed a biphasic response even 168 h after vaccination. The majority of the lipids were triglycerides (TAGs), where TAGs with long-chain unsaturated fatty acids (FAs) decreased at 24 h and TAGs with short-chain FAs decreased at 168 h. To our knowledge, this is the first report on the impact of human vaccine adjuvant Alum on the host metabolome, which may provide new insights into the mechanism of action of Alum.


Beneficial effects of Lactobacillus reuteri 6475 on bone density in male mice is dependent on lymphocytes.
Oral treatment with probiotic bacteria has been shown to prevent bone loss in multiple models of osteoporosis. In previous studies we demonstrated that oral administration of Lactobacillus reuteri in healthy male mice increases bone density. The host and bacterial mechanisms of these effects however are not well understood. The objective of this study was to understand the role of lymphocytes in mediating the beneficial effects of L. reuteri on bone health in male mice. We administered L. reuteri in drinking water for 4 weeks to wild type or Rag knockout (lack mature T and B lymphocytes) male mice. While L. reuteri treatment increased bone density in wild type, no significant increases were seen in Rag knockout mice, suggesting that lymphocytes are critical for mediating the beneficial effects of L. reuteri on bone density. To understand the effect of L. reuteri on lymphocytes in the intestinal tissues, we isolated mesenteric lymph node (MLN) from naïve wild type mice. In ex vivo studies using whole mesenteric lymph node (MLN) as well as CD3+ T-cells, we demonstrate that live L. reuteri and its secreted factors have concentration-dependent effects on the expression of cytokines, including anti-inflammatory cytokine IL-10. Fractionation studies identified that the active component of L. reuteri is likely water soluble and small in size (<3 kDa) and its effects on lymphocytes are negatively regulated by a RIP2 inhibitor, suggesting a role for NOD signaling. Finally, we show that T-cells from MLNs treated with L. reuteri supernatants, secrete factors that enhance osterix (transcription factor involved in osteoblast differentiation) expression in MC3T3-E1 osteoblasts. Together, these data suggest that L. reuteri secreted factors regulate T-lymphocytes which play an important role in mediating the beneficial effects of L. reuteri on bone density.


A model of postnatal formation of alveoli in rat lung.
In rats, and many other species, most lung alveoli are formed after birth. Septation of the large air saccules existing at birth has been considered as the main mechanism for alveoli formation. However, other undefined means of alveolarization have also been postulated to account for the large increase in gas-exchange surface area that takes place in the lung as the rat grows larger. Moreover, recent results show that the majority of alveoli in rat lung are formed by means other than septation of saccules existing at birth, but these mechanisms have not been identified up to the present. In this study, a mathematical model of alveolarization in rat lung is presented. The model is based on three postulates: (a) new saccules continue to be formed up to adulthood according to certain rules; (b) all these saccules subsequently septate generating a certain number of alveoli; (c) once formed, the saccules (and alveoli) do not change in volume, but newly-formed saccules are larger than the preceding ones according to a given law. The model accurately predicts the experimentally-known values at different ages of total alveolar volume, alveolar number, volume of the average alveolus, gas-exchange surface area, and alveolar volume distribution for normal rats and for rats in which septation is inhibited by treatment with dexamethasone or hypoxia during the early postnatal weeks of life.


Effect of cold-restraint stress on glutathione and lipid peroxide levels in the liver and glandular stomach of rats.
The effect of starvation and cold-restraint stress on glutathione and lipid peroxide levels in the liver, stomach and plasma of rats was investigated. Hepatic and gastric glutathione levels were significantly decreased in starvation and cold-restraint groups when compared with values obtained from the control group. In both tissues, lipid peroxide levels were significantly decreased after starvation but were not significantly different from control values after cold-restraint treatment. However, when compared with the values obtained from the starvation group there was a significant increase in both hepatic and gastric lipid peroxide levels after cold-restraint. Plasma lipid peroxide levels were slightly decreased in the starvation group and significantly increased in the cold-restraint group. Our results suggest that pathological consequences of stress on different tissues could be due to stimulation of lipid peroxidation.


Bulk protonic conductivity in a cephalopod structural protein.
Proton-conducting materials play a central role in many renewable energy and bioelectronics technologies, including fuel cells, batteries and sensors. Thus, much research effort has been expended to develop improved proton-conducting materials, such as ceramic oxides, solid acids, polymers and metal-organic frameworks. Within this context, bulk proton conductors from naturally occurring proteins have received somewhat less attention than other materials, which is surprising given the potential modularity, tunability and processability of protein-based materials. Here, we report proton conductivity for thin films composed of reflectin, a cephalopod structural protein. Bulk reflectin has a proton conductivity of ~2.6 × 10(-3) S cm(-1) at 65 °C, a proton transport activation energy of ~0.2 eV and a proton mobility of ~7 × 10(-3) cm(2) V(-1) s(-1). These figures of merit are similar to those reported for state-of-the-art artificial proton conductors and make it possible to use reflectin in protein-based protonic transistors. Our findings may hold implications for the next generation of biocompatible proton-conducting materials and protonic devices.


Association of cognitive impairment and breast cancer survivorship on quality of life in younger breast cancer survivors.
Younger breast cancer survivors (BCS) often report cognitive impairment and poor quality of life (QoL), which could be interrelated. The purpose of this study was to examine the association of cognitive impairment and breast cancer status (BCS versus healthy control (HC)), with QoL, which included psychological (depressive symptoms, well-being, perceived stress, and personal growth) and physical well-being (physical functioning and fatigue). Four hundred ninety-eight BCS (≤45 years at diagnosis) who were 3 to 8 years post-chemotherapy treatment and 394 HC completed subjective questionnaires and a one-time neuropsychological assessment, including tests of attention, memory, processing speed, and verbal fluency. For each test, cognitive impairment was defined as scoring 1.5 and 2.0 standard deviations below the mean of the HC group. Separate linear regression models for each outcome were ran controlling for known covariates. BCS reported significantly more memory problems than HC (p < 0.0001), with up to 23% having significant impairment. Cognitive performance did not differ significantly between BCS and HCs. BCS vs. HCs had greater depression and fatigue, yet more personal growth. Objective and subjective cognitive impairment were significantly related to greater depressive symptoms and perceived stress and lower well-being and physical functioning; whereas, objective impairment was related to less personal growth and subjective impairment was related to greater fatigue. Younger BCS report significant cognitive impairment years after treatment which may relate to greater decrements in QoL. Assessment and interventions to address cognitive concerns may also influence QoL outcomes in younger BCS.


In-vessel composting of HMX and RDX contaminated sludge using microbes isolated from contaminated site.
Current study was carried out with an objective to remediate highly contaminated sludge with HMX and RDX obtained from an explosive manufacturing facility in North India employing indigenous microbes, Arthrobacter subterraneus (isolate no. S2-TSB-17) and Bacillus sonorensis (isolate no. S8-TSB-4) which were isolated from the same contaminated site. In-vessel composting of the explosive contaminated sludge was performed in 12 different bioreactors using cow manure and garden waste as bulking agents. 78.5% degradation of HMX was observed in reactor no. 2 with Bacillus sonorensis having combination of 10% sludge, 70% cow manure and 20% garden waste on 80th day. Two secondary metabolites Bis(hydroxymethyl)nitramine and methylene dinitramine were identified while studying the degradation pathway. Similarly, degradation of 91.2% was observed for RDX in reactor no. 11 with consortia of Arthrobacter subterraneus and Bacillus sonorensis on 80th day. During the study, release of significant nitrate and nitrite ions were observed. It has already been established that RDX and HMX degradation leads to release of nitrite/nitrate ions. The highest nitrite (reactor no. 11) and nitrate (reactor no. 2) release observed were 24.02 ± 0.05 mg/kg and 30.65 ± 0.99 mg/kg on 50th and 70th day, respectively. Scanning electron microscopic studies confirmed the attachment and presence of microbes with solid surface and no deformation in structure was observed in the microbial cells due to contamination stress. Findings of the study concluded that in-vessel composting assisted with native bacterial species can be a potential technology for the treatment of explosive contaminated sludge at the contaminated sites.


Lung expansion, tidal exchange, and formation of the functional residual capacity during resuscitation of asphyxiated neonates.
Ventilatory exchange and endotracheal and esophageal pressures were measured during resuscitation of asphyxiated neonates born by cesarean section. In contrast to spontaneously breathing, vaginally born babies, an opening pressure had to be exceeded before lung expansion occurred. Subsequently there was usually a gradual increase in gaseous exchange over the first few lung inflations. A further rise in lung compliance occurred with the baby's inspiratory efforts. The functional residual capacity was formed with or without active inspiratory efforts by the baby, although gaseous retention occurred more rapidly as a result of the infant's inspiration.


Detection of antibodies to Toxoplasma gondii and Neospora caninum in animals from three zoos in Slovakia.
The aim of this study was to detect antibodies to Toxoplasma gondii and Neospora caninum in exotic animal species kept in three zoos in Slovakia. Antibodies to T. gondii and N. caninum were detected by commercial Enzyme-linked immunosorbent assay (ELISA, ID Screen Toxoplasmosis Indirect Multispecies and ID Screen Neospora caninum Indirect Multispecies, ID Vet, Montpellier, France). Antibodies to T. gondii and N. caninum were detected in 43% (24/56) and 5% (3/55) of animals, respectively. The three animals with N. caninum antibodies: two wolves (Canis lupus) and one Hartmann's mountain Zebra (Equus zebra hartmannae), were clinically healthy, and both wolves simultaneously had antibodies to T. gondii. The results of our study provide a picture of the recent circulation of T. gondii in three Slovakian zoos with the S/P (ratio of antibodies in the sample to antibodies in positive control) value higher than 200%, found in five animals (9%) indicating acute toxoplasmosis. The highest S/P value (296%) was detected in a Roloway monkey (Cercopithecus roloway), which was healthy without clinical signs, presuming that Roloway monkey is a species less susceptible to T. gondii infection. Results of our study showed the presence of T. gondii and N. caninum in Slovakian zoos, confirming recent T. gondii infections according to the high level of antibodies detected in five animals, referring to acute toxoplasmosis.


Interleukin-6, interleukin-8, and soluble tumor necrosis factor receptor-I in the cord blood as predictors of chronic lung disease in premature infants.
In order to predict the late-development of chronic lung disease of prematurity (CLD), cytokines in the cord blood were assessed in this study. Eighteen premature infants with CLD were enrolled. Cord blood plasma levels of cytokines of these infants and 12 control infants without CLD were measured including interleukin (IL)-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, soluble TNF receptor-I, and soluble IL-6 receptor using a cytometric bead array and an enzyme-linked immunosorbent assay. The cord blood IL-6, IL-8, and sTNFR-I levels were significantly elevated in CLD infants compared with those in control (P < .05). IL-1beta, IL-2, IL-4, IL-10, and IFN-gamma were undetectable in both groups. CLD infants with maternal chorioamnionitis had higher IL-6 than those without chorioamnionitis (P < .01). In CLD infants, IL-6 was higher in the infants who required prolonged oxygen therapy (P < .05). Elevated inflammatory cytokines in the cord blood are associated with the progression to CLD.


Prognostic significance of secondary cytogenetic changes and nonspecific cross-reacting antigen (NCA) in patients with Ph-positive chronic myeloid leukemia.
Cytogenetic analyses were carried out on peripheral blood and bone marrow cells of 31 chronic myeloid leukemia (CML) patients who presented with blastic, accelerated, or chronic phases. The percentage of cytoplasmic nonspecific cross-reacting antigen (cNCA, a marker of myelocytic differentiation)-containing cells was determined in the same blood or bone marrow samples. The patients were divided in two groups according to cytogenetic results: those with aberrations in addition to the Philadelphia chromosome (Ph1) and those with Ph1 only. Among the additional aberrations such changes, not typical of CML, were found: del(2)(p21), t(6;11)(q25;q23), and t(12;?)(p13;?). The survival time and the percentage of cNCA-positive cells of patients in blastic and accelerated phases were compared between the above-mentioned two groups of patients using the Student t test and the Kaplan-Meier estimator. The percentage of cNCA-positive cells was significantly lower and the survival time significantly shorter in the group of patients with additional aberrations. The probability of survival according to the Kaplan-Meier estimator was also lower for this group. These data suggest that the immunologically determined lower degree of maturity, that characterized cells bearing additional aberrations, coincides with and/or results in more rapid progression of the disease.


A complex translocation event between the two homologues of chromosomes 5 leading to a del(5)(q21q33) as a sole aberration in a case clinically diagnosed as CML: characterization of the aberration by multicolor banding.
We report on a patient with a clinically diagnosed Philadelphia negative chronic myelogenous leukemia (CML) with a so far unrecorded complex translocation event between the two homologue chromosomes 5. At the GTG-band level the karyotype was normal, apart from an enlarged chromosome 5 and an extremely shortened second chromosome 5. Both derivative chromosomes 5 consisted exclusively of #5 derived material as proven by 24-color FISH. To characterize the complex aberration in more detail the multicolor banding (MCB) technique using a chromosome 5 specific probe set was applied. Using this DNA-based high resolution banding procedure, the karyotype could be described as 46,XX,del(5)(pterright curved arrow q12::q33right curved arrow qter),ins(5)(pterright curved arrow q15::q12right curved arrow q21::q21right curved arrow qter). In consequence, the aberration leads to a partial deletion of the long arm of chromosome 5: del(5)(q21q33), which would not have been identified using conventional banding techniques or 24-color FISH.


Effects of magnesium sulfate on an unfolding step of human cyanomet myoglobin.
The effects of magnesium sulfate (MgSO4) on an unfolding step of human cyanomet myoglobin (Mb) were examined for wild-type and three L-->A mutant Mbs. The unfolding was induced at acidic pH (3.6-4.5) with various concentrations of MgSO4 (0-2 M). The monophasic process was monitored by visible absorption spectroscopy. We observed quite nonlinear delta G not equal to-[MgSO4] relations for all the Mbs. delta G not equal to-[MgCl2] relations were also determined for a comparative study. Thermodynamic evaluation of the results indicated that an upward reflection of delta G not equal to-[MgSO4] relations in high [MgSO4] is caused by the strong Hofmeister effect of the salt. Results obtained for three mutants (L29A, L72A, and L104A) at pH 4.0 and 4.5 were consistent with our previous observation that the structure of the transition state is determined by the stability of Mb cores in the balance with the pH conditions of unfolding (T. Konno and I. Morishima. 1993. Biochim. Biophys. Acta. 1162:93-98).


Injury trends in alpine skiing and a snowboarding over the decade 2008-09 to 2017-18.
This research explores snowsport injury trends in western Canadian resorts over the decade 2008-2018. Ecological study. Injury and participation data on alpine skiing and snowboarding was provided by the Canada West Ski Areas Association for 2008-2018. Inclusion criteria were met by 29 of 52 resorts, resulting in the analysis of 107,540 injury reports via SPSS 24, using descriptive statistics and chi-squared analysis. Over the decade: there was a gradual decline in the injury rate, averaging 2.24 injuries per 1000 skier days. Male snowboarders accounted for 1/3 of all injured participants, females were 42% of all. Day-ticket holders were the most injured of all customer-types, with most injuries occurring as the result of falls on marked, green/easiest terrain. For skiers, injuries to lower limbs, and especially knees, were most common. For snowboarders, upper limb injuries, especially wrists, were most common. 12.8% of the injured population required emergency transport. Advanced slopes/terrain had the highest proportion requiring emergency transport. Fewer skiers experienced head injuries than snowboarders. Snowboarders experienced a higher proportion of serious injuries. Snowsport injuries are relatively rare when compared to other sports. Skiers were most prone to knee injuries and snowboarders to wrist injuries. Emergency transport was proportionally needed more from advanced slopes/terrain. Further research is needed into why and how participants make decisions around risk. Future injury prevention strategies should be evaluated in light of potential risk factors such as, social determinants of injury, fall mechanisms, changing participation patterns, age, snowsport discipline, and legal cannabis use.


Streptococcus pluranimalium: A novel human pathogen?
We present the first case of a subdural empyema caused by Streptococcus pluranimalium, in a healthy adolescent male as a possible complication of subclinical frontal sinusitis. Clinical features, diagnostic approach and management of subdural empyema are discussed. A 17-year-old male with a 2 day history of headache and nausea was referred to our Emergency Department (ED) as a case of possible meningitis. He was afebrile, lethargic and drowsy with significant neck stiffness on examination. Computerized tomography (CT) revealed a large frontotemporoparietal subdural fluid collection with significant midline shift. Subsequent contrast-enhanced CT established the presence of intracranial empyema; the patient underwent immediate burr-hole evacuation of the pus and received 7 weeks of intravenous antibiotics, recovering with no residual neurological deficit. The diagnosis of subdural empyema as a complication of asymptomatic sinusitis in an immunocompetent patient with no history of fever or upper respiratory symptoms was unanticipated. Furthermore, the organism Streptococcus pluranimalium that was cultured from the pus has only been documented twice previously in medical literature to cause infection in humans, as it is primarily a pathogen responsible for infection in bovine and avian species. Subdural empyema represents a neurosurgical emergency and if left untreated is invariably fatal. Rapid diagnosis, surgical intervention and intensive antibiotic therapy improve both morbidity and mortality.


A FORTRAN computer program for storage and retrieval of personal journal references.
A FORTRAN program is described for interactive maintenance of a personal library of journal references and reprints. The program queries the user for category (author, journal, year, title, and keyword) information during entry of new documents. Retrieval of documents in the set is accomplished by entering as many search words as desired, in any order, and in any category. The program returns documents that match for all entered words. Other functions include terminal display of the information stored for a given document and the ability to generate an alphabetized bibliographic listing from among the document set. A correction routine and a routine for listing the contents of the search word files are also included.


pilG Gene cluster and split pilL genes involved in pilus biogenesis, motility and genetic transformation in the cyanobacterium Synechocystis sp. PCC 6803.
The unicellular motile cyanobacterium Synechocystis sp. PCC 6803 exhibits phototactic motility that depends on the type IV-like thick pilus structure. By gene disruption analysis, we showed that a gene cluster of slr1041, slr1042, slr1043 and slr1044, whose predicted products are homologous to PatA, CheY, CheW and MCP, respectively, was more or less required for pilus assembly, motility and natural transformation competency with extraneous DNA. By sequence homology, the missing cheA-like gene in this cluster was identified as novel split genes, slr0073 and slr0322, at separate loci on the genome. This was confirmed by non-motile phenotype of their disruptants. Unique hyperpiliation was observed in the slr1042 and slr0073 disruptants, suggestive of their specific interaction with pilT1. The genes, thus identified as pil genes in this study, were designated pilG (slr1041), pilH (slr1042), pilI (slr1043), pilJ (slr1044), pilL-N (slr0073) and pilL-C (slr0322).


CCN2: a bona fide target for anti-fibrotic drug intervention.
CCN2 (formerly known as connective tissue growth factor) was identified by several different laboratories approximately 20 years ago. Almost since its identification as a factor induced in normal fibroblasts by transforming growth factor β and overexpressed in fibrotic disease, CCN2 has been hypothesized to be not only a marker but also a central mediator of fibrosis in vivo. Finally, in vivo data are emerging to validate this key hypothesis. For example, a neutralizing anti-CCN2 antibody was found to attenuate fibrogenesis in three separate animal models (Wang et al. in Fibrogenesis Tissue Repair 4:1-4, 2011). This commentary addresses recent data indicating that CCN2 appears to represent a key central mediator of fibrosis and a good target for anti-fibrotic drug intervention.


Assessment of Patients' Willingness to Participate in a Randomized Trial of Spinal versus General Anesthesia for Lumbar Spine Surgery.
Prospective nonrandomized studies have found less postoperative fatigue and improved quality of life in patients undergoing awake spine surgery under spinal anesthesia compared with general anesthesia. Randomized trials are needed to validate these findings. To prospectively investigate patients' willingness to enroll in randomized trials of lumbar spine surgery under spinal versus general anesthesia and identify any potential barriers. We recruited patients undergoing lumbar spine surgery for degenerative disease. We described a randomized trial of spine surgery under spinal versus general anesthesia and assessed patients' willingness to participate in such trial. We elicited preferences for treatment along with demographics. The association between these factors and willingness to participate in the trial was examined. Fifty patients completed interviews; 58% were female, mean age of 60.9 ± 12.5 years. A total of 52% patients stated that they were definitely willing to participate in the hypothetical randomized trial, and 8% probably willing. Only 16% of patients were aware of spinal anesthesia as an option for low back surgery, and 60% indicated no strong preference for the anesthesia techniques. Patients without strong preferences stated a greater willingness to participate than those with strong preferences (80% vs. 10% definitely willing, P < 0.0001). Age, sex, education, work status, and race were not significantly associated with willingness to participate. Sixty percent of patients stated that they were either definitely or probably willing to participate in the randomized trial. Subjects lacking strong preferences for the anesthesia technique stated a greater willingness to enroll than those with strong preference.


Modified use of the proximal humeral internal locking system (PHILOS) plate for distal femoral nonunions.
Nonunion is a common complication after a distal femoral fracture (DFF). Standard treatment consists of revision plating and/or bone grafting. Single lateral plating for a distal femoral nonunion can be insufficient in case of a persistent medial gap and compromised bone stock. Alternatively, dual plating can be used to treat a distal femoral nonunion, but to date there is no Gold standard. The aim of our study was to report our results after use of a minimally invasively placed proximal humeral internal locking system (Philos) plate as a medial buttress in the treatment of a distal femoral nonunion. Fifteen adult patients with a distal femoral nonunion were prospectively entered in a trauma database and retrospectively assessed. All patients underwent a similar operation, which included removal of failed hardware, nonunion debridement, fixation with a lateral plate, and a medial Philos plate combined with bone grafting. Data collected included union rate, time to union, complications and functional outcome. In twelve out of fifteen patients (80%), the fracture united after our index operation. Median time to union was 4.8 months (range 1.6-15). Three patients (20%) needed additional bone grafting surgery. One patient underwent a Judet quadricepsplasty. This study suggests that the Philos plate is a safe and effective adjunct as a medial buttress plate for distal femoral nonunions.


Expression of Kaposi's sarcoma-associated herpesvirus-encoded K10/10.1 protein in tissues and its interaction with poly(A)-binding protein.
The K10/10.1 protein is encoded by a cluster of interferon regulatory factor (IRF) homologues in the Kaposi's sarcoma-associated herpesvirus (KSHV, human herpesvirus 8, HHV-8) genome. In the present study, we showed that an anti-K10 antibody reacted with a 110-kDa protein encoded by the K10/10.1 gene of KSHV in KSHV-infected primary effusion lymphoma (PEL) cell lines. Expression of K10/10.1 protein was induced by phorbol ester in KSHV-infected cells. A reporter gene assay demonstrated that K10/10.1 protein did not influence promoter activity of human interferon genes, regardless of its homology to human IRFs. Poly(A)-binding protein (PABP) was identified as a partner of K10/10.1 protein. Immunoprecipitation revealed that K10/10.1 protein interacted with PABP specifically in PEL cell lines. IFA revealed co-localization of K10/10.1 protein and PABP in the nucleus of KSHV-infected cells. These data suggest that K10/10.1 protein may affect the translational status or stability of mRNA in host cells.


Drug interactions with antiretrovirals.
The recent development of new antiretroviral drugs, along with the evolution in clinical practice guidelines that include the recommendation of the use of three- to four-drug combination regimens for achieving optimal suppression of viral replication, has focused clinicians on the relevance of drug-drug interactions in the chronic care of HIV-infected individuals. However, the routine clinical management of drug interactions is complicated by our expanding knowledge of the physiologic mechanisms underlying pharmacokinetic interactions, particularly as they relate to drug transport and tissue distribution (eg, P-glycoprotein) and biotransformation (hepatic cytochrome p450 mono-oxygenase induction and inhibition). This review provides an updated summary of key drug interactions that have been reported since its initial publication.


Results of renal replacement therapy in Europe, 1980 to 1987. Registration Committee of the EDTA-ERA.
The characteristics of the patient populations accepted for and undergoing renal replacement therapy (RRT) in Europe in 1980 to 1987 are presented. Acceptance rates have increased in most countries reporting to the EDTA Registry and have reached from between 50 to over 80 per million population in the more affluent Western European countries in 1987. Increasing acceptance rates were due to the inclusion of patient groups at a higher risk of dying, such as the elderly and those with diabetic nephropathy. Despite the acceptance of a growing proportion of high-risk patients, no increase in overall mortality was apparent. Gross mortality (some 10% annually) changed little between 1980 and 1987 for patients on hemodialysis, decreased sharply from 1980 to 1984 for patients on continuous ambulatory peritoneal dialysis (CAPD), and improved continuously from 4.2% in 1980 to 2.4% in 1987 for patients with a functioning kidney graft. In order to determine mortality more accurately, actuarial survival rates and annual death rates per thousand patient years at risk were computed according to age groups and to primary renal disease groups, both for the total patient data file and for selected countries. Actuarial 5-year survival on hemodialysis for all patients starting treatment between 1982 and 1987 varied according to age, being 84% in patients age 15 to 24 years and 20% in those age 75 to 84 years. Patients with "standard" primary renal diseases had slightly better survival, while of the group with diabetic nephropathy only 51% age 25 to 34 and no more than 3% of those age 75 to 84 survived the 5-year mark. To recognize trends in the mortality between 1980 and 1987, annual death rates for all patients on record, age 45 to 54 and 55 to 64 at the beginning of 1980, 1983, 1985, and 1987 were computed both for the total Registry and for the FRG. Despite an increasing acceptance rate of patients with diabetic nephropathy, the annual death rates on hemodialysis decreased or remained stable both for the total Registry and for the FRG. Death rates in patients with a functioning graft decreased. It is concluded that, during the last decade, survival on RRT has continued to improve not only because of decreasing mortality after transplantation and on CAPD, but also due to improving survival on hemodialysis. The latter is not readily apparent because of the increasing acceptance rate in older patient groups and a rapidly rising proportion of patients with diabetic nephropathy in most European countries.


Laparoscopic and Robotic Management of Ureteral Stricture in Adults.
The aim of this review was to provide an update on the status of minimal invasive treatment of ureteral stricture either with a laparoscopic or robotic surgery. Eligible studies, published until November 2019 were retrieved through Medline, Cochrane and Pubmed databases. Predetermined inclusion and exclusion criteria were used as selection method for data synthesis and acquisition. The study was performed in accordance with the PRISMA statement. A total of 19 retrospective studies met the inclusion criteria. All of them demonstrated the safety, feasibility and success of both laparoscopic and robotic ureteral reconstruction. Individual case series or cumulative comparison analysis of the available studies showed at least equivalent success rates and a trend favoring laparoscopic and robotic groups in terms of estimated blood loss and length of hospital stay to the detriment of longer operative times and possibly higher cost. Current evidence suggests the effectiveness, safety and increasing incorporation of minimally invasive techniques for complex stricture repair and reconstruction.


Tortuosity Index Predicts Early Successful Reperfusion and Affects Functional Status After Thrombectomy for Stroke.
The literature is scarce for studies evaluating the anatomy of cervical vessels in patients with stroke. We sought to investigate the effect of vessel tortuosity in procedural, angiographic, and functional outcomes in patients with acute ischemic stroke treated with mechanical thrombectomy (MT). Patients with an emergent large vessel occlusion of the anterior circulation treated with MT between 2015 and 2020 were included. The tortuosity of the internal carotid artery was recorded as the tortuosity index (TI) using the following formula: [(actual/straight length-1) × 100). A multivariable regression was performed to assess procedural, angiographic, and functional outcomes based on the TI. A total of 212 patients were included. Median age was 72 years (interquartile range, 62-82 years); admission National Institutes of Health Stroke Scale score was 17 ± 6. Median TI was 7.9 (interquartile range, 3.7-19.7). A total of 127 patients (60%) had a TI <10. Early reperfusion (procedure time <60 minutes) was accomplished in 144 patients (67.9%). A multivariable analysis showed that patients with a TI <10 were more likely to achieve an early reperfusion (odds ratio [OR], 2.3; 95% confidence interval [CI], 1.11-4.78; P = 0.025). A TI <10 was a predictor of successful reperfusion (OR, 2.0; CI, 1.05-3.93; P = 0.035) and an early reperfusion was the sole predictor of functional independence (most recent modified Rankin Scale score 0-2) (OR, 4.1; 95% CI, 1.62-10.53; adjusted P = 0.003). Patients with a TI <10 are significantly more likely to achieve early successful reperfusion after MT for the treatment of acute ischemic stroke.


Mutagenic activities of gentisin and isogentisin from Gentianae radix (Gentianaceae).
The mutagenic activities of 2 hydroxyxanthones, gentisin and isogentisin, obtained from the methanol extract of Gentianae radix (Gentianaceae) were investigated. The methanol extract of Gentianae radix, which showed mutagenicity in the Ames test in Salmonella typhimurium strain TA100 with S9 mix, was fractionated by column chromatography on Sephadex LH-20, and the fractions were purified by preparative TLC and column chromatography on polyamide. 2 mutagenic materials thus obtained, S1 and S2, each gave a single band on TLC. Identification of S1 and S2 was accomplished by comparing the analytical (mps, elementary analyses) and spectral (UV, IR, mass, NMR) results for S1 and S2 with literature data for gentisin and isogentisin. At doses below 10 micrograms, S1 (gentisin) and S2 (isogentisin) had similar specific mutagenic activities. At doses of over 10 to 50 micrograms, the mutagenic activities of S2 and S1 were 19.1 and 6.94 revertants per microgram respectively. This much lower activity of S1 than S2 may be a result of its poor solubility owing to the presence of the OMe group at C-3. The combined yield of S1 and S2 was about 76 mg (40 mg as S1 and 36 mg as S2), which accounted for 76% of the content of mutagenic compounds (100 mg) estimated roughly from the total mutagenic activity in the extract of the starting materials (100 g).


Evaluation of progressive hepatic histopathology in long-term tamoxifen therapy.
Tamoxifen (TAM) therapy is the better treatment for breast cancer and the drug use the prophylaxis of this disease in young premenopausal women. Yet, the effects associated with this therapy are unknown. To better understand the extension of this problem, we developed an animal model to mimic this therapy, aiming to evaluate its potential biochemical and histopathological changes in the liver. Young cycling female rats were treated with TAM for one, two and three months and toxicological biomarkers and liver histomorphometry were evaluated. Starting at two months, TAM-treatment prevented the normal age-dependent increase in body weight, without inducing changes in food intake. Serum levels of cholesterol and of the metabolic enzymes creatine kinase and aspartate aminotransferase were reduced in all TAM treatment periods. Serum levels of the metabolic enzymes alkaline phosphatase and lactate dehydrogenase were increased after the first month but returned to control levels upon 3 months of drug exposition. Moderate microvesicular steatosis, classified only at the first month of TAM treatment, was reduced afterwards. Our model showed an adaptive response of liver upon 3 months of treatment, suggesting that at the stated conditions, TAM will not promote hepatotoxicity. In this way, the present model may be useful in the study of possible key endocrine effects of TAM use and the search for better clinical outcomes.


Analysis of mnemonic processing by means of totally reversible neural inactivations.
The irreversible lesions technique precludes the analysis of the possibly critical role played by discrete brain sites in the several distinct stages of mnemonic processing (acquisition, consolidation, retrieval) during which these may be specifically but transiently active. On the contrary, the reversible functional inactivation techniques, by means of stereotaxic local microinjection of active compounds, make it possible to suppress the neuronal function of a discrete volume of nervous tissue, for a pre-determined time, with the assurance of complete functional recovery within a known duration. This technique makes it possible to block the neural activity of a chosen neural site at a given stage of memory processing without any interference with the function of the same structure either during earlier or later stages of the same process. Thus, the reversible ablation results may provide information not only on the qualitative topographical but also on the quantitative temporal dimension of learning and memory. The technique employed to cause totally reversible neural inactivation is detailed. The employment of several agents to obtain functional inactivation is discussed. Of these, perhaps the safest and most manageable is tetrodotoxin when a fairly long functional inactivation (e.g., 1 h) is desired. The effects of a reversible inactivation can be quite easily and accurately assessed by observing the severity of the amnesic disruption, if any, of a conditioned response. In order to do this as well as possible, it is advantageous to employ a very simple behavioral paradigm. The passive avoidance response in the light-dark box apparatus fulfills this requirement. Moreover, this paradigm, being one-trial, provides the necessary condition of a single well-defined temporal beginning. The present protocol has been successfully employed in learning and memory research, to assess when the functional integrity of a given neural structure is necessary in order that a conditioned response may be acquired, consolidated or retrieved. The employment of this protocol in relation to the intrinsic functional characteristics of a given subcortical neural site is discussed.


Methodological optimization of polyethylenimine (PEI)-based gene delivery to the lungs of mice via aerosol application.
Polyethylenimine (PEI) has been successfully used for gene delivery to the lungs of mice via aerosol application using a whole body nebulization device. In this report we optimized the design of such an aerosol device. Aerosol devices were constructed as either serial inhalation apparatus or as a whole body nebulization chamber connected to an aerosol spacer placed in a horizontal or vertical position. PEI-based gene vectors were nebulized using a standard jet nebulizer and luciferase gene expression of various tissues was examined. Using a whole body aerosol device resulted in luciferase gene expression in the lungs of mice at the same level as compared with a serial inhalation apparatus. Whereas gene expression was enhanced in the presence of 5% CO(2)-in-air, anesthesia of mice strongly decreased gene expression even when mice were intubated with an intravascular cannula. Reduction of the median mass aerodynamic diameter (MMAD) of the aerosol from 3.4 to 0.27 microm by interposition of an aerosol spacer increased gene expression significantly 3-fold. Drying of the aerosol by silica gel additionally increased gene delivery significantly 3-fold. Reporter gene expression mediated by branched PEI 25 kDa was 9- and 15-fold higher as compared with linear PEIs of 22 and 25 kDa, respectively, and was dependent on the DNA concentration. Gene expression was detectable as soon as 6 h after gene vector application and reached a maximum after 72 h but was still detectable after 14 days. The presence of Zn(2+) did not increase gene expression. We propose aerosol drying as a novel and simple method of optimizing PEI-based gene delivery to the lungs.


Antibiotic resistance patterns of Acinetobacter calcoaceticus-A. baumannii complex species from Colombian hospitals.
Only automated phenotypic methods are currently used in Colombian hospitals for identifying isolates of the Acinetobacter calcoaceticus-A. baumannii complex (ACB). The phenotypical similarities in these species mean that they cannot be differentiated by manual or automated methods, thereby leading to their identification as A. baumannii, or ACB complex in clinical settings. Our objective was to identify to the species level 60 isolates, from four hospitals, evaluate their antibiotic susceptibility, and detect resistance-related genes. 16S-23S rRNA internal transcribed spacer (ITS) region and rpoB gene partial sequences were amplified. Resistance genes for cephalosporin, carbapenem and aminoglycoside were detected by PCR. Possible mutations in the quinolone resistance-determining region (QRDR) were evaluated. The association of ISAba-1 with blaOXA and blaADC genes was determined by PCR. Amplification products of ITS region, rpoB gene and some resistance genes were sequenced and compared using the BLAST tool. 16S-23S rRNA ITS region and partial rpoB gene sequence analysis allowed 51isolates to be identified as A. baumannii, 8 as A. nosocomialis, and 1 isolate as A. pitti. A. baumannii isolates were highly resistant to all antibiotics tested, while the others were susceptible to ciprofloxacin and ampicillin/sulbactam. Quinolone resistance, found only in A. baumannii, was associated with mutations in the QRDR region of gyrA and parC genes. This is the first investigation in Colombia that has identified ACB complex species using molecular methods, and determined differences in antibiotic resistance and resistance genes among the species. It is of the highest importance to identify isolates to the species level for future resistance and epidemiology studies in our region.


Neural labeling and manipulation by neonatal intraventricular viral injection in mice.
This step-by-step protocol provides a fast and easy technique to label and/or genetically manipulate neural cells, achieved by intraventricular injection of viral vectors into neonatal mice under ultrasound guidance. Successful injection of adeno-associated viral vectors (AAV) induces neural transduction as fast as 3 days post injection (dpi) in both the central and peripheral nervous systems. Virally driven expression persists until early adulthood. The same setup enables injection of other viral vectors as well as intramuscular injection. For complete details on the use and execution of this protocol, please refer to Wang et al. (2021) and Brill et al. (2016).


1-[4-(3-{[5-(4-Chloro-phen-yl)furan-2-yl]methyl-idene-amino}-2,5-dioxoimidazol-idin-1-yl)but-yl]-4-methyl-piperazine-1,4-diium dichloride hemihydrate.
The title compound, C(23)H(30)ClN(5)O(3) (2+)·2Cl(-)·0.5H(2)O, was synthesized by N-alkyl-ation of 1-({[5-(4-chloro-phen-yl)-2-furan-yl]methyl-ene}amino)-2,4-imidazolidinedione with 1-bromo-4-chloro-butane, and N-methyl-piperazine. In the crystal, the cations, anions and water mol-ecules are linked by O-H⋯Cl and N-H⋯Cl hydrogen bonds.