Transdiaphragmatic tunneled Broviac catheters: Cost-effective perioperative central venous access in infants undergoing cardiac surgery.
Infants undergoing congenital heart surgery require central venous lines which can be achieved by various combinations of transthoracic lines, percutaneous-indwelling central catheters and tunneled Broviac catheters. Transthoracic lines are removed by protocol prior to cardiac intensive care unit discharge (risk of bleeding), at which time percutaneous-indwelling central catheters are placed. Transdiaphragmatic tunneled Broviac catheters placed at the time of sternotomy, remain in place until hospital discharge, when they are safely removed at bedside. We characterized actual cost profiles associated with strategies that do versus do not include tunneled Broviac catheters. From January 2014 to December 2016, we identified a study population of 220 consecutive patients under 1 year of age undergoing congenital heart surgery. Cost data were acquired from our electronic patient system interface database and office of finance. Our cohort was divided into 2 groups, tunneled Broviac catheter and nontunneled Broviac catheter. We calculated the total cost associated with each groups' central venous lines, propensity matched, and used the Mann-Whitney U test to analyze the results. Eighty-three (37.7%) of the 220 patients had tunneled Broviac catheters. The tunneled Broviac catheter group had 4 percutaneous-indwelling central catheter insertions and 6 radiological interventions while the nontunneled Broviac catheter group had 90 percutaneous-indwelling central catheters and 203 radiologic interventions. After propensity score matching, both groups were reduced to 82 patients and sum, median and interquartile range cost for tunneled Broviac catheters and nontunneled Broviac catheters was $17,351.84, $159.76 (128-159.76) versus $72,809.32, $1277.26 (31.76-1277.26), P < .02 respectively. Tunneled Broviac catheters, placed routinely at cardiac surgery, incur lower costs than the conventional combination of transthoracic lines and percutaneous-indwelling central catheters. The cost-effectiveness is achieved by reducing the number of percutaneous-indwelling central catheters and associated radiologic interventions.


Synthesis and evaluation of a new inhibitor of phosphoglucose isomerases: the enediolate analogue 5-phospho-D-arabinohydroxamate.
Designed as a high energy intermediate analogue inhibitor of the potent chemotherapeutic target phosphoglucose isomerases, 5-phospho-D-arabinohydroxamate was efficiently synthesized in a two steps procedure. To date, it proved to be the strongest competitive inhibitor with respect to substrate D-fructose-6-phosphate (Ki down to 98 nM and Km/Ki values up to 513). A comparative inhibition study of this compound and other known strong inhibitors on phosphoglucose isomerases from three different sources is also reported.


Machine Learning Algorithms for Classification of First-Trimester Fetal Brain Ultrasound Images.
To evaluate the feasibility of machine learning (ML) tools for segmenting and classifying first-trimester fetal brain ultrasound images. Two image segmentation methods processed high-resolution fetal brain images obtained during the nuchal translucency scan: "Statistical Region Merging" (SRM) and "Trainable Weka Segmentation" (TWS), with training and testing sets in the latter. Measurement of the fetal cerebral cortex in original and processed images served to evaluate the performance of the algorithms. Mean absolute percentage error (MAPE) was used as an accuracy index of the segmentation processing. The SRM plugin revealed a total MAPE of 1.71% ± 1.62 SD (standard deviation) and a MAPE of 1.4% ± 1.32 SD and 2.72% ± 2.21 SD for the normal and increased NT groups, respectively. The TWS plugin displayed a MAPE of 1.71% ± 0.59 SD (testing set). There were no significant differences between the training and testing sets after 5-fold cross-validation. The images obtained from normal NT fetuses and increased NT fetuses revealed a MAPE of 1.52% ± 1.02 SD and 2.63% ± 1.98 SD. Our study demonstrates the feasibility of using ML algorithms to classify first-trimester fetal brain ultrasound images and lay the foundation for earlier diagnosis of fetal brain abnormalities.


An overview of carbapenemase producing enterobacteriaceae (CPE) in trauma and orthopaedics.
Carbapenemase resistant enterobacteriae (CPE) may be found in asymptomatic carriers. Its incidence is increasing worldwide. Surgical patients are at increased risk of immunocompromise and of carriage progressing to active infection. Active infection with CPE caries a high mortality rate, with the bacteria being resistant to many antibiotics. This article provides details on the epidemiology, screening and management of the orthopaedic patient with CPE. The guidelines advise orthopaedic staff on ways to avoid the spread of CPE amongst inpatients.


Ganglionated plexus ablation vs linear ablation in patients undergoing pulmonary vein isolation for persistent/long-standing persistent atrial fibrillation: a randomized comparison.
The optimal ablation technique for persistent and long-standing persistent atrial fibrillation (AF) is unclear. Both linear lesion (LL) and ganglionated plexus (GP) ablation have been used in addition to pulmonary vein isolation (PVI), but no direct comparison of the 2 methods exists. The aim of this study is to assess the comparative safety and efficacy of 2 different ablation strategies-PVI+LL vs PVI+GP ablation -in patients with persistent or long-standing persistent AF. Two hundred sixty-four consecutive patients with persistent/long-standing persistent AF were randomly assigned to 2 different ablation schemes: PVI+LL (n = 132) and PVI+GP (n = 132) ablation. Consistent sinus rhythm (SR) off antiarrhythmic drug was assessed after follow-up of at least 3 years with the use of an implanted monitoring device. All procedural end points were acutely achieved. At 12 months after a single procedure, 47% of the patients treated with PVI+LL were in SR compared to 54% of the patients treated with PVI+GP (P = .29). At 3 years, 34% of the patients with PVI+LL and 49% of the patients with PVI+GP maintained SR (P = .035). Atrial flutter was more frequent in the PVI+LL group than in PVI+GP group (18% vs 6%; P = .002). After a second procedure in 78 patients of the PVI+LL group and 55 patients of the PVI+GP group, the long-term overall success rate was 52% and 68%, respectively (P = .006). PVI+GP ablation confers superior clinical results with less ablation-related left atrial flutter and reduced AF recurrence compared to PVI+LL ablation at 3 years of follow-up.


Vaginal prostaglandin E2 for interruption of pregnancy and management of intrauterine death.
Vaginal suppositories containing 20 mg prostaglanding E2, administered at 2-5h intervals, are very effective in interrupting second-trimester pregnancy and in inducing labor in case of death in utero. However, side effects are common and make the treatment unpleasant to most patients. Premedication with an maintenance of a potent anti-emetic (e.g. haloperidol) and an anti-diarrheic (e.g. loperamide) considerably reduce the frequency and severity of these side effects.


A nanoreactor boosts chemodynamic therapy and ferroptosis for synergistic cancer therapy using molecular amplifier dihydroartemisinin.
Chemodynamic therapy (CDT) relying on intracellular iron ions and H2O2 is a promising therapeutic strategy due to its tumor selectivity, which is limited by the not enough metal ions or H2O2 supply of tumor microenvironment. Herein, we presented an efficient CDT strategy based on Chinese herbal monomer-dihydroartemisinin (DHA) as a substitute for the H2O2 and recruiter of iron ions to amplify greatly the reactive oxygen species (ROS) generation for synergetic CDT-ferroptosis therapy. The DHA@MIL-101 nanoreactor was prepared and characterized firstly. This nanoreactor degraded under the acid tumor microenvironment, thereby releasing DHA and iron ions. Subsequent experiments demonstrated DHA@MIL-101 significantly increased intracellular iron ions through collapsed nanoreactor and recruitment effect of DHA, further generating ROS thereupon. Meanwhile, ROS production introduced ferroptosis by depleting glutathione (GSH), inactivating glutathione peroxidase 4 (GPX4), leading to lipid peroxide (LPO) accumulation. Furthermore, DHA also acted as an efficient ferroptosis molecular amplifier by direct inhibiting GPX4. The resulting ROS and LPO caused DNA and mitochondria damage to induce apoptosis of malignant cells. Finally, in vivo outcomes evidenced that DHA@MIL-101 nanoreactor exhibited prominent anti-cancer efficacy with minimal systemic toxicity. In summary, DHA@MIL-101 nanoreactor boosts CDT and ferroptosis for synergistic cancer therapy by molecular amplifier DHA. This work provides a novel and effective approach for synergistic CDT-ferroptosis with Chinese herbal monomer-DHA and Nanomedicine.


Transcriptome analysis of thermomorphogenesis in ovules and during early seed development in Brassica napus.
Plant sexual reproduction is highly sensitive to elevated ambient temperatures, impacting seed development and production. We previously phenotyped this effect on three rapeseed cultivars (DH12075, Topas DH4079, and Westar). This work describes the transcriptional response associated with the phenotypic changes induced by heat stress during early seed development in Brassica napus. We compared the differential transcriptional response in unfertilized ovules and seeds bearing embryos at 8-cell and globular developmental stages of the three cultivars exposed to high temperatures. We identified that all tissues and cultivars shared a common transcriptional response with the upregulation of genes linked to heat stress, protein folding and binding to heat shock proteins, and the downregulation of cell metabolism. The comparative analysis identified an enrichment for a response to reactive oxygen species (ROS) in the heat-tolerant cultivar Topas, correlating with the phenotypic changes. The highest heat-induced transcriptional response in Topas seeds was detected for genes encoding various peroxidases, temperature-induced lipocalin (TIL1), or protein SAG21/LEA5. On the contrary, the transcriptional response in the two heat-sensitive cultivars, DH12075 and Westar, was characterized by heat-induced cellular damages with the upregulation of genes involved in the photosynthesis and plant hormone signaling pathways. Particularly, the TIFY/JAZ genes involved in jasmonate signaling were induced by stress, specifically in ovules of heat-sensitive cultivars. Using a weighted gene co-expression network analysis (WGCNA), we identified key modules and hub genes involved in the heat stress response in studied tissues of either heat-tolerant or sensitive cultivars. Our transcriptional analysis complements a previous phenotyping analysis by characterizing the growth response to elevated temperatures during early seed development and reveals the molecular mechanisms underlying the phenotypic response. The results demonstrated that response to ROS, seed photosynthesis, and hormonal regulation might be the critical factors for stress tolerance in oilseed rape.


Can we prevent type 1 diabetes?
Type 1 diabetes (T1D) affects 1.93 in 1000 youth in the USA. Over the last 40 years, a combination of genetic and immunological markers has been developed allowing for the accurate prediction of progression to T1D. Despite our abilities to predict disease and the marked improvement in our understanding of the natural history of T1D, therapies capable of preventing or reversing T1D remain elusive. This article will review recent and ongoing efforts to understand the causes of T1D and related efforts to study potential therapies aimed at preventing T1D.


Ethanol inhibits hormone stimulated hepatocyte DNA synthesis.
Insulin, glucagon, and epidermal growth factor (EGF) addition stimulated DNA synthesis in primary hepatocyte cell cultures prepared from adult rat liver. The addition of ethanol (20-200mM) to the culture medium resulted in a substantial reduction in DNA synthesis as measured by 3H-thymidine incorporation and autoradiography. This effect was specific for differentiated hepatocytes compared to fibroblasts and two other human hepatoma cell lines. These studies demonstrate in a cell culture system that one of the major properties of ethanol is the inhibition of hepatocyte DNA synthesis.


Dementia-related disability in the population aged 90 years and over: differences over time and the role of comorbidity in the vitality 90 + study.
The burden of dementia, multimorbidity, and disability is high in the oldest old. However, the contribution of dementia and comorbidities to functional ability in this age group remains unclear. We examined the combined effects of dementia and comorbidities on ADL and mobility disability and differences between dementia-related disability between 2001, 2010, and 2018. Our data came from three repeated cross-sectional surveys in the population aged 90 + in the Finnish Vitality 90 + Study. The associations of dementia with disability and the combined effects of dementia and comorbidity on disability adjusted for age, gender, occupational class, number of chronic conditions, and study year were determined by generalized estimating equations. An interaction term was calculated to assess differences in the effects of dementia on disability over time. In people with dementia, the odds of ADL disability were almost five-fold compared to people with three other diseases but no dementia. Among those with dementia, comorbidities did not increase ADL disability but did increase mobility disability. Differences in disability between people with and without dementia were greater in 2010 and 2018 than in 2001. We found a widening gap in disability between people with and without dementia over time as functional ability improved mainly in people without dementia. Dementia was the main driver of disability and among those with dementia, comorbidities were associated with mobility disability but not with ADL disability. These results imply the need for strategies to maintain functioning and for clinical updates, rehabilitative services, care planning, and capacity building among care providers.


The origin structure of each finger in the flexor digitorum superficialis muscle.
The flexor digitorum superficialis muscle (FDS) is considered the most important of the forearm flexors for maintaining elbow valgus stability. However, the relationships between the origin structure of each finger of the FDS and the anterior oblique ligament (AOL) of the ulnar collateral ligament and the common tendon (CT) in the proximal part, and morphological features are unclear. The purpose of this study was to clarify the relationships between the origin structure of each finger of the FDS and the AOL and the CT, as well as to clarify the morphological features of the muscle belly of each finger of the FDS. This study examined 20 elbows. The origin of each finger was examined. Muscle mass, muscle fiber bundle length, and the pennation angle of each finger were also measured. In all cases, the third and fourth digits originated from the radius, the anterior common tendon (ACT), and the posterior common tendon (PCT). The second and fifth digits (18 elbows) or an independent fifth digit (2 elbows) originated from the ACT, the PCT, the AOL, and other soft tissues of the elbow. Muscle mass and muscle fiber bundle length in the muscle belly of the third and fourth digits were significantly heavier and longer, respectively, than in the muscle belly of the second and fifth digits. Because the second and fifth digits or an independent fifth digit originated from the AOL, their contraction may cause tension in the AOL.


HNO induces DNA deletions in the yeast S. cerevisiae.
HNO is genotoxic but its mechanism is not well understood. There are many possible mechanisms by which HNO can attack DNA. Since HNO is electrophilic, it may react with exocyclic amine groups on DNA bases and through a series of subsequent reactions form a deaminated product. Alternatively, HNO may induce radical chemistry through O(2)-dependent (or possibly O(2)-independent) chemistry. In cell free systems, experiments have shown that HNO does react with DNA, resulting in base oxidation and strand cleavage. In this study, we used a whole-cell system in the yeast Saccharomyces cerevisiae to study the mechanism of HNO induced DNA damage with Angeli's salt as HNO donor. The yeast DEL assay provided a measure of intrachromosomal recombination leading to DNA deletions. We also examined interchromosomal recombination leading to genomic rearrangements and used the canavanine (CAN) assay to study induction of forward point mutations. HNO was a potent inducer of DNA deletions and recombination but it was negative for induction of point mutations. This suggests that HNO causes DNA strand breaks rather than base damage. Genotoxicity was observed under aerobic and anaerobic conditions and NAC protected against HNO induced DNA deletions. Since HNO is genotoxic under anaerobic conditions, NAC probably protected against radicals generated by HNO independent of oxygen.


An analytical solution for ground water transit time through unconfined aquifers.
An exact, closed-form analytical solution is developed for calculating ground water transit times within Dupuit-type flow systems. The solution applies to steady-state, saturated flow through an unconfined, horizontal aquifer recharged by surface infiltration and discharging to a downgradient fixed-head boundary. The upgradient boundary can represent, using the same equation, a no-flow boundary or a fixed head. The approach is unique for calculating travel times because it makes no a priori assumptions regarding the limit of the water table rise with respect to the minimum saturated aquifer thickness. The computed travel times are verified against a numerical model, and examples are provided, which show that the predicted travel times can be on the order of nine times longer relative to existing analytical solutions.


Functional analysis of evolutionary conserved clustering of bZIP binding sites in the baculovirus homologous regions (hrs) suggests a cooperativity between host and viral transcription factors.
The genome of the Autographa californica Multinucleocapsid Polyhedrosis Virus (AcMNPV) contains nine interspersed homologous regions (hrs) that function as potent enhancer sequences when linked in cis to either viral or heterologous RNA polymerase II-dependent promoters. Their activity is strongly increased by the binding of the major immediate early viral transregulator IE1 on 28-mer palindromic sites present in hrs. We show that hrs of AcMNPV additionally carry, in the interpalindromic sequences, a large number of cAMP response elements (CRE) and TPA response elements (TRE), known to bind ubiquitous cellular transcription factors of the bZIP family. Moreover, these clusters of CRE and TRE motifs are concentrated in hrs. Analysis of the 25 baculovirus genomes sequenced so far reveals that these motifs are evolutionary conserved in Lepidoptera NPVs, suggesting a functional role in the hr enhancer function. Consistently, EMSA experiments indicate that CRE and on a lesser extent TRE sites specifically bind insect host factors. Moreover, reporter assays reveal that these CRE sites have an additive stimulatory effect on RNAPol II-dependent transcription in Sf9 cells and are potentially able to synergize with the IE1-binding palindrome.


Safety and prognostic role of regional lymphadenectomy for primary and metastatic liver tumors.
Routine regional lymphadenectomy for colorectal liver metastases and primary liver tumors is still a subject for debate. During 2001-2005, we performed a prospective study of cases in which regional lymphadenectomy around the hepato-duodenal ligament and common hepatic artery was applied (group R+ LN) or not (group R- LN). Pre-operative clinical features of patients were comparable among groups as well as the operative data. There were 108 (67%) males; the median age was 66 years; 124 cases had a single lesion (77%), and the median diameter was 4 cm. The type of lesion was: 77 (48%) colorectal liver metastases (M-CR), 75 (46%) hepatocellular carcinomas (HCC) and 10 (6%) cholangiocellular carcinomas. In the R+ LN group, the mean number of lymph nodes removed was 6.7 ± 4.8 (range 4-26), and seven cases (8.6%) presented lymph node metastasis. The median follow-up was 3.5 years. M-CR patients showed comparable hospital mortality (R+ LN 0% vs. R- LN 2.6%) and morbidity (R+ LN 17.9% vs. R- LN 21.1%), but R+ LN had higher 5-year disease-free survival (31 vs. 16%, p < 0.05). HCC cases in the R+ LN group presented higher hospital mortality (13.5 vs. 0%, p < 0.05) without any improvement in disease-free survival (it was at 5-year disease-free survival 34 vs. 33%, respectively, p = n.s.). Routine regional lymphadenectomy should be performed for colorectal liver metastases, and avoided in patients with hepatocellular carcinoma.


The use of historical sources in a multi-layered methodology for karez research in Turpan, China.
In this article, we will present an overview of possible research methods to handle historical sources, in the specific case of karez landscapes. A karez system is an underground water collection system, prevalent in the Turpan basin of China. Sources and the associated methodology have become more important today, because of contemporary issues such as modernisation, urbanisation and agricultural expansion. These problems make it harder to read the landscape, which is why we have to start extracting our data from maps, reports, photographs, and satellite imagery. We will give a short overview of sources, each with an explanation of their processing method. Despite certain cautions that should be taken into account, these methods clearly complement the current state of knowledge on the Turpan karez. As this paper is part of a special issue, Water History in the time of COVID-19, it has undergone modified peer review.


The adenoid as a key factor in upper airway infections.
The adenoids (and the nasopharynx) play a key role in the normal functioning and in various pathologies of the upper respiratory tract. In this paper the role of adenoidal pathology and the beneficial effect of adenoidectomy in some upper respiratory tract and facial anomalies and diseases are discussed; otitis media with effusion, recurrent acute otitis media, sinusitis, snoring and sleep apnea and abnormal patterns in the midface growth and development.


PBMC MicroRNAs: Promising Biomarkers for the Differential Diagnosis of COVID-19 Patients with Abnormal Coagulation Indices.
MicroRNAs, or miRNAs, may involve in coagulation and inflammation pathways caused by severe Coronavirus disease (COVID-19). Accordingly, this attempt was made to explore the behavior of peripheral blood mononuclear cells (PBMCs) miRNAs as effective biomarkers to diagnose COVID-19 patients with normal and abnormal coagulation indices. We selected the targeted miRNAs (miR-19a-3p, miR-223-3p, miR-143-5p, miR-494-3p and miR-301a-5p) according to previous reports, whose PBMC levels were then determined by real-time PCR. Receiver operating characteristic (ROC) curve was obtained to clarify the diagnostic potency of studied miRNAs. The differentially expressed miRNA profiles and corresponding biological activities were predicted in accordance with bioinformatics data. Targeted miRNAs' expression profiles displayed a significant difference between COVID-19 subjects with normal and abnormal coagulation indices. Moreover, the average miR-223-3p level expressed in COVID-19 cases with normal coagulation indices was significantly lower than that in healthy controls. Based on data from ROC analysis, miR-223-3p and miR-494-3p are promising biomarkers to distinguish the COVID-19 cases with normal or abnormal coagulation indices. Bioinformatics data highlighted the prominent role of selected miRNAs in the inflammation and TGF-beta signaling pathway. The differences existed in the expression profiles of selected miRNAs between the groups introduced miR-494-3p and miR-223-3p as potent biomarkers to prognosis the incidence of COVID-19.


Operating room ventilation with laminar airflow shows no protective effect on the surgical site infection rate in orthopedic and abdominal surgery.
To evaluate whether operating room (OR) ventilation with (vertical) laminar airflow impacts on surgical site infection (SSI) rates. Retrospective cohort-study based on routine surveillance data. Sixty-three surgical departments participating voluntarily in the German national nosocomial infections surveillance system "KISS" were included (a total of 99,230 operations). Active SSI surveillance was performed according to the methods and definitions given by the US National Nosocomial Infection Surveillance system. Surgical departments were stratified according to type of OR ventilation used: (1) turbulent ventilation with high-efficiency particulate air-filtered air, and (2) HEPA-filtered (vertical) laminar airflow ventilation. Multivariate analyses were performed by the generalized estimating equations method to control for the following variables as possible confounders: (a) Patient-based: wound contamination class, ASA score, operation duration, patients' age and gender, endoscopic operation; (b) Hospital-based: the number of beds in the hospital, its academic status, operation frequency, and long-term participation in KISS. The risk for severe SSI after hip prosthesis implantation was significantly higher using laminar airflow OR ventilation (1.63 < 1.06; 2.52>), as compared with turbulent ventilation. The adjusted odds ratios for the other operative procedures analyzed were: knee prosthesis 1.76 < 0.80, 3.85>; appendectomy 1.52 < 0.91, 2.53>; cholecystectomy 1.37 < 0.63, 2.97>; colon surgery 0.85 < 0.49, 1.49>; and herniorrhaphy 1.48 < 0.67; 3.25>. Unexpectedly, in this analysis, which controlled for many patient and hospital-based confounders, OR ventilation with laminar airflow showed no benefit and was even associated with a significantly higher risk for severe SSI after hip prosthesis.


The biological effect of metal ions on the granulation of aerobic granular activated sludge.
As a special biofilm structure, microbial attachment is believed to play an important role in the granulation of aerobic granular activated sludge (AGAS). This experiment was to investigate the biological effect of Ca(2+), Mg(2+), Cu(2+), Fe(2+), Zn(2+), and K(+) which are the most common ions present in biological wastewater treatment systems, on the microbial attachment of AGAS and flocculent activated sludge (FAS), from which AGAS is always derived, in order to provide a new strategy for the rapid cultivation and stability control of AGAS. The result showed that attachment biomass of AGAS was about 300% higher than that of FAS without the addition of metal ions. Different metal ions had different effects on the process of microbial attachment. FAS and AGAS reacted differently to the metal ions as well, and in fact, AGAS was more sensitive to the metal ions. Specifically, Ca(2+), Mg(2+), and K(+) could increase the microbial attachment ability of both AGAS and FAS under appropriate concentrations, Cu(2+), Fe(2+), and Zn(2+) were also beneficial to the microbial attachment of FAS at low concentrations, but Cu(2+), Fe(2+), and Zn(2+) greatly inhibited the attachment process of AGAS even at extremely low concentrations. In addition, the acylated homoserine lactone (AHL)-based quorum sensing system, the content of extracellular polymeric substances and the relative hydrophobicity of the sludges were greatly influenced by metal ions. As all these parameters had close relationships with the microbial attachment process, the microbial attachment may be affected by changes of these parameters.


An online-based intervention to promote healthy eating through self-regulation among children: study protocol for a randomized controlled trial.
Despite the enormous investment governments allocate to fight obesity, its worldwide prevalence is still on the rise. Moreover, the majority of the programs implemented are still targeting adults struggling with overweightness and focusing on transmitting knowledge about food. However, research shows that obesity prevention is more efficacious and cheaper, and beliefs about healthy eating have a stronger influence on eating behavior than declarative knowledge about food. In fact, knowledge about healthy eating only influences weight status when combined with self-regulation competences. Thus, the main goal of the current project is to develop and evaluate the efficacy of an online preventive intervention program, the HEP-S. This program is designed to promote and develop a set of transversal skills and strategies, related to self-regulation, on the healthy eating domain among school-aged children. A three-armed randomized controlled trial will be conducted in several schools in Portugal. It will include a standard control group, with no intervention; an online intervention group, with the program for 20 weeks; and an enhanced online intervention group, with the program for 20 weeks embedded with gamification strategies throughout the program. Per research group, 40 groups of about 15 children each will be recruited and measured at five different time points. The three research groups will complete the same assessment protocol at the same timings (baseline, post-intervention, and 3, 6, and 9 months' follow-ups). The assessment protocol will include anthropometric and psychological measures. The primary outcome measures will be the development of self-regulation skills for healthy eating over time, the development of self-efficacy attitudes, knowledge about healthy eating over time, and others. The secondary outcome measures will include the effect of gamification strategies, engagement, and satisfaction with the program, among others. The program will comprise the following: (i) a weekly group synchronous videoconference session with a trained educational psychologist serving as a mediator and (ii) a weekly parental involvement activity. Narratives, or story-tools, embedded with self-regulation strategies are at the core of the intervention. The program may play an important role in preventing risky and unhealthy eating behaviors by focusing on the development of self-regulation skills and strategies among elementary school children. ClinicalTrials.gov NCT04099498 . Registered on 23 September 2019.


Differential roles of protease isoforms in the tumor microenvironment.
Alternative splicing of precursor mRNA is a key mediator of gene expression regulation leading to greater diversity of the proteome in complex organisms. Systematic sequencing of the human genome and transcriptome has led to our understanding of how alternative splicing of critical genes leads to multiple pathological conditions such as cancer. For many years, proteases were known only for their roles as proteolytic enzymes, acting to regulate/process proteins associated with diverse cellular functions. However, the differential expression and altered function of various protease isoforms, such as (i) anti-apoptotic activities, (ii) mediating intercellular adhesion, and (iii) modifying the extracellular matrix, are evidence of their specific contribution towards shaping the tumor microenvironment. Revealing the alternative splicing of protease genes and characterization of their protein products/isoforms with distinct and opposing functions creates a platform to understand how protease isoforms contribute to specific cancer hallmarks. Here, in this review, we address cancer-specific isoforms produced by the alternative splicing of proteases and their distinctive roles in the tumor microenvironment.


The effect of nectar production on neighborhood size.
Bumblebee foraging behavior was observed on two plant species with similar floral and inflorescence structures. One species produces nectar while the other does not. Bees, upon visiting nectar producing flowers tend to empty them of nectar and by frequently moving between close neighbors, create a patchily distributed resource base. Bees maximize their foraging efficiency in such an environment by using an area-restricted searching behavior and flying distances inversely correlated with the quality of reward received. Pollen collecting bumblebees do not create a patchy environment and maximize their foraging efficiency by more consistently moving shorter distances. Pollen collecting bumblebees are significantly more likely to revisit flowers and to visit more flowers per inflorescence than are nectar gathering bumblebees. These differences in foraging behavior increase the neighborhood size for nectar producing species and make it increasingly unlikely that random drift will be a dominant mode of evolution in populations of these species.


Phenotype and function of dendritic cells and T-lymphocyte polarization in the human colonic mucosa and adenocarcinoma.
To evaluate the status of activation of the intestinal dendritic cells (DCs) and T lymphocytes (T cells) from surgical specimens of human colon and adenocarcinoma, and the potential effect of administration of interleukin 2 (IL-2). Patients undergoing colectomy for cancer were randomized to receive subcutaneous IL-2 (12million UI/day) (treated group; n=10) for 3days before operation or no treatment (control group, n=10). DCs and T cells were isolated and purified from the lamina propria (LP) of segments of normal colon and adenocarcinoma of both groups. Cell phenotype was determined by expression of membrane receptors. Interaction between DC and T cells was assesses by a mixed leukocyte reaction using naïve T cells co-cultured with DCs. CD4+ T-cell polarization was studied by intracellular staining with monoclonal antibodies for interleukin-4 and interferon-gamma. CD4+ T cells were significantly less in tumour than in LP (p<0.05) in both treated and control groups. IL-2 did not modify the number of any of the T-cell subsets analysed. In contrast, T cells isolated from LP and neoplasm of treated patients produced more interferon-gamma and less interleukin-4 (p<0.05 vs. controls). IL-2 administration significantly increased (p<0.05) the number of mature, myeloid and plasmocytoid DCs compared to controls. Allogeneic naïve T cells were polarized toward a Th1 type of response which appeared to be mediated by IL-2 activated DCs. systemic IL-2 treatment may have immunomodulatory properties on intestinal DC maturation and drive a Th1 mediated anti-neoplastic response.


Latent (slowly progressing) autoimmune diabetes in adults.
About 10% of patients with the clinical presentation of type 2 diabetes suffer from an autoimmune form of diabetes associated with a rapid decline of residual beta-cell mass and subsequent development of insulin dependency. In this condition, called latent autoimmune diabetes in adults (LADA), there are clinical and metabolic features intermediate between type 1 and type 2 diabetes. Recent studies provide novel information on the immune markers associated with progressive beta-cell loss in LADA patients. However, LADA pathogenesis is still poorly understood; further studies are needed to establish general recommendation for preventing and treating this subtype of autoimmune diabetes.


Cloning of caf1+, caf2+ and caf4+ from Schizosaccharomyces pombe: their involvement in multidrug resistance, UV and pH sensitivity.
We previously identified four nuclear genes (caf1+ to caf4+) in Schizosaccharomyces pombe, mutations in which can confer caffeine resistance. Here we report the cloning and sequencing of caf1+, caf2+ and caf4+. All three genes are allelic to genes (hba1+, crm1+ and trr1+, respectively) involved in multidrug resistance mechanisms or in stress response systems. In agreement with this the caffeine-resistant mutants caf1(hba1)-21, caf2(crm1)-3 and caf4(trr1)-83 are also resistant to brefeldin. Disruption of caf1(hba1)+ and caf4(trr1)+ makes cells sensitive to high pH. The overlapping ranges of pleiotropic effects and the genetic interaction detected between caf1(hba1)+ and caf2(crm1)+ suggest that the three genes function in interlinked systems.


CTBP1‑AS upregulation is associated with polycystic ovary syndrome and can be effectively downregulated by cryptotanshinone.
Polycystic ovary syndrome (PCOS) is one of the most common endocrinopathies and primarily presents with hyperandrogenism. Although environmental factors and genetic factors are thought to be the major reason, there still exists a lot of questions need to be answered. High expression of C‑terminal‑binding protein 1 antisense (CTBP1‑AS) was identified as an independent risk factor for PCOS; however, the molecular mechanism of CTBP1‑AS in PCOS regulation is unknown. In the present study, the expression level of CTBP1‑AS was found to be significantly upregulated in patients with PCOS compared with healthy control patients. CTBP1‑AS knockdown was demonstrated to reduce the proliferation and promote the apoptosis of granulosa tumor cells in vitro. It was also identified that the two core catalytic subunits of Polycomb repressive complex 2 (enhancer of zeste homolog 2 and embryonic and ectoderm development protein) interacted with CTBP1‑AS in primary granulosa cells and KGN cells. In addition, cryptotanshinone treatment was demonstrated to effectively downregulate CTBP1‑AS expression level. Data from the present study suggested a pathophysiological role of CTBP1‑AS in PCOS and may provide a new potential target for PCOS treatment.


Focal polymicrogyria, continuous spike-and-wave discharges during slow-wave sleep and Cohen syndrome: a case report.
Cohen syndrome is a rare genetic disorder consisting of truncal obesity, hypotonia, mental retardation, characteristic facial appearance and ocular anomalies. Other diagnostic clinical features include narrow hands and feet, low growth parameters, neutropenia and chorioretinal dystrophy. Here, we report an 18-year-old male with Cohen syndrome associated with focal polymicrogyria and continuous spike-and-wave discharges during slow-wave sleep.


Comparative analysis of the effects of synthetic derivatives of batrachotoxin on sodium currents in frog node of Ranvier.
1. In voltage-clamp experiments on frog myelinated nerve fibers, the effects of nine synthetic derivatives of batrachotoxin (BTX) obtained from 7,8-dihydrobatrachotoxinin A (DBTX-A) on Na+ currents (INa) have been investigated. 2. Both of 20 alpha-esters of DBTX-A with 2,4,5-trimethylpyrrol-3-carboxylic acid (DBTX-P) and benzoic acid (DBTX) at a 10(-5) M concentration caused modification of INa qualitatively similar to that induced by BTX. 3. The quaternary derivative of DBTX (QDBTX) produced such changes in INa only at a 5.10(-4) M concentration, apparently due to its much lower lipid solubility. 4. Replacement of a -CH2- by a -C = O. group in the homomorpholine ring near the tertiary nitrogen atom abolished the DBTX activity, strongly suggesting the necessity of tertiary nitrogen protonation for the toxin interaction with the channel receptor. 5. Transfer of an 11-hydroxygroup from the alpha- to the beta-position in the DBTX molecule did not decrease its activity in spite of the fact that in the beta-position this group is sterically very hindered. The activity of 11 beta-DBTX is at variance with the prediction of Codding's (1983) "oxygen triad" hypothesis. 6. DBTX-A and compounds obtained from DBTX by oxidation of the 11 alpha-hydroxygroup (K-DBTX), acetylation (Ac-DBTX), or reduction of the hemiketal moiety (H2DBTX) even at a concentration as high as 10(-3) M were able to modify only a very small fraction of the Na channels. However, a clear-cut reversible blocking action on both normal and modified Na channels was observed. 7. These results led us to conclude that BTX modifies the Na channels only in the charged form and hemiketal and 20 alpha-ester moieties provide adequate disposition of toxin on the receptor surface. The inability of H2DBTX, DBTX-A, and K-DBTX and Ac-DBTX to modify most of the Na channels can be explained by a low "probability of correct disposition" of these ligands on the receptor surface.


A designed fusion tag for soluble expression and selective separation of extracellular domains of fibroblast growth factor receptors.
Fibroblast growth factor receptors (FGFRs) generate various transduction signals by interaction with fibroblast growth factors (FGFs) and are involved in various biological functions such as cell proliferation, migration, and differentiation. Malfunction of these proteins may lead to the development of various diseases, including cancer. Accordingly, FGFRs are considered an alternative therapeutic target for protein and/or gene therapy. However, the screening of antagonists or agonists of FGFRs is challenging due to their complex structural features associated with protein expression. Herein, we conducted the development of a protease-free cleavable tag (PFCT) for enhancing the solubility of difficult-to express protein by combining maltose-binding protein (MBP) and the C-terminal region of Npu intein. To validate the availability of the resulting tag for the functional production of extracellular domains of FGFRs (Ec_FGFRs), we performed fusion of PFCT with the N-terminus of Ec_FGFRs and analyzed the expression patterns. Almost all PFCT-Ec_FGFR fusion proteins were mainly detected in the soluble fraction except for Ec_FGFR4. Upon addition of the N-terminal region of Npu intein, approximately 85% of the PFCT-Ec_FGFRs was separated into PFCT and Ec_FGFR via intein-mediated cleavage. Additionally, the structural integrity of Ec_FGFR was confirmed by affinity purification using heparin column. Taken together, our study demonstrated that the PFCT could be used for soluble expression and selective separation of Ec_FGFRs.


Metabolism of adenine nucleotides in the cultured fetal mouse heart.
Intact beating fetal mouse hearts in organ culture were deprived of oxygen and glucose for up to 4 h, resulting in loss of beating, an 80% fall in ATP, reduction of energy charge from 0.85 to 0.48, and doubling of total nucleoside concentration. Radiolabeled adenine nucleotides were degraded to hypoxanthine and inosine, which were lost from the hearts into the medium during the deprivation period. Adenosine and adenine also appeared in the medium when adenosine deaminase was inhibited. After 24 h of O2 and glucose resupply, ATP returned to 60% of control, and energy charge rose to 0.76. Labeled nucleosides and bases remaining in the heart or exogenous labeled adenine were utilized to resynthesize ATP. [14C]glycine was rapidly taken up by recovering hearts but was not used for de novo adenine nucleotide synthesis. Ability to recover ATP and spontaneous contraction appear related to residual nucleotide and nucleoside content rather than to energy charge.


Improvement of gluten-free steamed bread quality by partial substitution of rice flour with powder of Apios americana tuber.
This study investigated the effect of powder made from tubers of the legume Apios americana (Apios) as a rice flour substitute in the making of gluten-free steamed bread. The carbohydrates of Apios powder were mainly starch and sucrose, and included legume-specific raffinose and stachyose. Apios powder contained almost no α-amylase but had a high level of β-amylase activity. Substitution of rice flour with Apios powder delayed the hardening of bread on storage and helped to maintain cohesiveness. Apios powder-substituted bread had higher maltose content than unsubstituted control bread due to β-amylase activity in the Apios powder. Bread substituted with 10% Apios powder had a significantly higher degree of gelatinization than the control even after storage, most likely due to lower amounts of recrystallized amylose as determined by differential scanning calorimetry. These results demonstrate Apios powder as promising a new food ingredient for improving the quality of gluten-free rice bread.


Detection of Chronic Wasting Disease Prions in Fetal Tissues of Free-Ranging White-Tailed Deer.
The transmission of chronic wasting disease (CWD) has largely been attributed to contact with infectious prions shed in excretions (saliva, urine, feces, blood) by direct animal-to-animal exposure or indirect contact with the environment. Less-well studied has been the role that mother-to-offspring transmission may play in the facile transmission of CWD, and whether mother-to-offspring transmission before birth may contribute to the extensive spread of CWD. We thereby focused on a population of free-ranging white-tailed deer from West Virginia, USA, in which CWD has been detected. Fetal tissues, ranging from 113 to 158 days of gestation, were harvested from the uteri of CWD+ dams in the asymptomatic phase of infection. Using serial protein misfolding amplification (sPMCA), we detected evidence of prion seeds in 7 of 14 fetuses (50%) from 7 of 9 pregnancies (78%), with the earliest detection at 113 gestational days. This is the first report of CWD detection in free ranging white-tailed deer fetal tissues. Further investigation within cervid populations across North America will help define the role and impact of mother-to-offspring vertical transmission of CWD.


Concurrent BRAFV600E and BRCA Mutations in MSS Metastatic Colorectal Cancer: Prevalence and Case Series of mCRC patients with prolonged OS.
BRAF V600E+ microsatellite stable (MSS) metastatic colorectal cancer (mCRC) patients comprise up to 10% of advanced CRC. They have a poor prognosis with a median survival typically <1 year. Despite use of multi-agent 1st line chemotherapy regimens and combination targeted therapies, outcomes are still poor. In our Institutional Molecular Tumor Board (MTB) database, we identified 3 mCRC patients with MSS/BRAF V600E who also had a BRCA1 or BRCA2 co-mutation and had relatively long overall survivals. Prior studies suggested that BRCA mutations are uncommon in CRC and we queried the Foundation Medicine (FM) genomic database to evaluate the prevalence of these cases as well as those with co-mutations in other homologous recombination genes. 36,966 CRC pts were sequenced by FMI using hybrid capture comprehensive genomic profiling (CGP) to evaluate all classes of genomic alterations (GA) for pathogenic BRAF mutations and/or a mutation in BRCA1/2 or a co-mutation in other homologous recombination (HR) genes (BARD1, CDK12, FANCL, PALB2, ATM, RAD54L, CHEK2, BRAF, BRIP1, RAD51D, RAD51C, RAD51B, CHEK1). Selected cohort analysis of BRAF V600E co-mutated with BRCA1 and BRCA2 were separated into MSI-H and MSS cohorts. The clinicopathological features and genomic loss of heterozygosity (gLOH) of those with a BRAF V600E and a BRCA1/BRCA2 mutation were collected and analyzed. We also describe 3 consecutive cases of mCRC patients, identified through the Inova Schar Cancer Institute (ISCI) MTB registry, whom had prolonged OS. Of 36,966 colorectal cancer pts, 6.6% were BRAF V600E+ and 1.5% had any co-occurring HR gene mutation(s) with 0.6% of the total mCRC population having co-ocurring BRAF V600E and BRCA1/2 alterations. BRCA co-mutations were higher in MSI-High BRAF V600E, however 24.1% of co-occurrences were observed in MSS samples. BRCA1 co-mutation was more commonly associated with MSS BRAF V600E and was associated with a higher gLOH than MSI-H BRAF V600E (18.7% vs 2.8%; p <0.001). In our institutional MTB database, (3/241;1.2%) CRC patients were MSS, BRAF V600E+ with BRCA1 or BRCA2 co-mutations, all somatic in origin, with an average gLOH of 21.4% and overall survivals of 72+(alive), 17+(alive), and 30 months, respectively. Co-existence of BRAF V600E/BRCA1/2 may represent a unique subset of advanced MSS CRC that may have a better prognosis and represent an opportunity to test novel targeted therapies. The elevated gLOH in these cases may also be a valuable biomarker for these pts. Larger prospective clinical validation trials in this subset is warranted.


Renal clear cancer cells regulate the differentiation of urine-derived stem cells to carcinoma-associated fibroblasts via RUNX3/TGF-β1 signaling axis.
Clear cell renal cell carcinoma (ccRCC), the most common pathological subtype of renal cancer, is one of the significant health concerns due to limited clinically effective treatments. Nevertheless, targeting carcinoma-associated fibroblasts in the tumor microenvironment has emerged as a promising innovative strategy for renal cancer therapy. Thus, this study is aimed to explore the role and molecular mechanism of urine-derived stem cells (USCs) in the progression and metastasis of ccRCC. Initially, wound-healing and transwell experiments were used to assess the migration and invasion abilities of the cells. Then, western blot analysis (WB) and quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) analyses were used to demonstrate the relevant protein and messenger RNA expression levels. Finally, hematoxylin-eosin and immunohistochemical stainings were performed to evaluate metastasis and protein expression in lung tumors. The coculture of USCs with the ccRCC cell lines significantly enhanced their migratory and invasive abilities. WB and qRT-PCR analyses exhibited that ccRCC cell lines significantly increased cell mobility markers transcriptional and protein levels in USCs. Finally, the in vivo investigations in nude mice showed that USCs promoted the proliferation and migration of ccRCC-based xenograft tumors. In summary, these findings demonstrated that USCs promoted ccRCC tumorigenesis and development in vivo and in vitro by regulating the Runt-related transcription factor 3/transforming growth factor-β1 signaling axis.


Bringing door-to-needle times within the European benchmarks results in better stroke patients outcomes in a spoke hospital from the Apulian Region.
Door-to-needle time (DNT) is a key factor in acute stroke treatment success. We retrospectively analysed the effects of a new protocol aimed at reducing treatment delays in our single-centre observational series over a 1-year period (from October 1st 2021 to September 30th 2022). The time frame was divided into two semesters as a new protocol was started at the beginning of the second semester to ensure a rapid evaluation, imaging, and intravenous thrombolysis in all stroke patients attending our spoke-hospital serving 200,000 inhabitants. Logistics and outcome measures were obtained for each patient and compared before and after implementation of the new protocol. A total of 215 patients with ischemic stroke attended our hospital within a 1-year period (109 in the first semester, 96 in the second semester). Seventeen percent and 21% of all patients underwent acute stroke thrombolysis in the first and second semesters, respectively. DNTs were strongly reduced in the second semester (from 90 to 55 min), bringing this value below the Italian and European benchmarks. This resulted in better short-term outcomes (an average of 20%) as measured by both Δ NIHSS scores at 24 h and at discharge with respect to baseline.


Post-operative guidelines following hip arthroscopy.
Rehabilitation following hip arthroscopy can vary significantly. Existing programs have been developed as a collaborative effort between physicians and rehabilitation specialists. The evolution of protocol advancement has relied upon feedback from patients, therapists and observable outcomes. Although reports of the first femoroacetabular impingement (FAI) surgeries were reported in the 1930's, it was not until recently that more structured, physiologically based guidelines have been developed and executed. Four phases have been developed in this guideline based on functional and healing milestones achieved which allow the patient to progress to the next level of activity. The goal of Phase I, the protective phase, is to progressively regain 75% of full range of motion (ROM) and normalize gait while respecting the healing process. The primary goal of Phase II is for the patient to gain function and independence in daily activities without discomfort. Rehabilitation goals include uncompensated step up/down on an 8 inch box, as well as, adequate pelvic control during low demand exercises. Phase III goals strive to accomplish pain free, non-compensated recreational activities and higher demand work functions. Manual muscle testing (MMT) grading of 5/5 should be achieved for all hip girdle musculature and an ability to dynamically control body weight in space. Phase IV requires the patient be independent with home and gym programs and be asymptomatic and pain free following workouts. Return to running may be commenced at the 12 week mark, but the proceeding requirements must be achieved. Athletes undergoing the procedure may have an accelerated timetable, based on the underlying pathology. Recognizing the patient's pre-operative health status and post-operative physical demands will direct both the program design and the program timetable.


Effects of d,l-propranolol on apomorphine induced stereotyped behavior in rats.
The effects of d,l propranolol on stereotyped behavior induced by apomorphine administration was studied quantitatively. d,l Propranolol caused a 2.5 leftward displacement of the control dose-response curve constructed to apomorphine-induced stereotypy; the ED50 for apomorphine was reduced from 1.87 +/- 0.38 to 0.74 +/- 0.17. The potentiation was dependent on both d,l-propranolol and apomorphine doses. Results are discussed in the light of a possible interference of the central noradrenergic systems with the expression of dopaminergic-induced stereotyped behavior.


Peptide specific expansion of CD8(+) T cells by recombinant plate bound MHC/peptide complexes.
Development of methods for efficient in vitro stimulation and expansion of peptide specific CD8(+) T cells is compelling not only with respect to adoptive T cell therapy but also regarding analysis of T cell responses and search for new immunogenic peptides. In the present study, a new approach to in vitro T cell stimulation was investigated. By use of an antigenic peptide derived from the cytomegalovirus (CMVp) we tested the stimulatory efficacy of recombinant plate bound MHC molecules (PB-MHC), being immobilized in culture plates. A single stimulation of non-adherent peripheral blood mononuclear cells (NA-PBMCs) with PB-MHC/CMVp resulted in significant expansion of CMVp specific CD8(+) T cells, which was comparable to that achieved by CMVp pulsed mature dendritic cells (DCs). By repeated exposure of NA-PBMCs to PB-MHC/CMVp more than 60% CMVp specific CD8(+) T cells, representing a 240-fold expansion, were reached after only two stimulations. Although stimulation with PB-MHC/CMVp clearly demonstrated efficient peptide specific expansion of CD8(+) T cells, there was a tendency to proliferative exhaustion of the cells after 3-4 stimulations. Thus, it will be of interest to examine the effect of new stimulatory cocktails, e.g. cytokines and co-stimulatory molecules, by use of the present rapid and easy-to-use method of expanding peptide specific T cells.


A temporary decrease in twitch response following reversal of rocuronium-induced neuromuscular block with a small dose of sugammadex in a pediatric patient.
We report a temporary decrease in twitch response following reversal of rocuronium-induced neuromuscular block with a small dose of sugammadex in our dose-finding study in pediatric patients. A 19-month-old female infant (9.6 kg, 80 cm) was scheduled for elective cheiloplasty surgery. Anesthesia was induced with nitrous oxide 50% and sevoflurane 5% and maintained with air, oxygen, sevoflurane 3%, and fentanyl (total, 3 μg/kg). Neuromuscular monitoring was performed at the adductor pollicis muscle after induction of anesthesia but before the administration of rocuronium. Total dose of rocuronium during the surgery was 0.9 mg/kg. Neuromuscular block was reversed with 0.5 mg/kg sugammadex when one response was observed with post-tetanic count stimulation. Twitch responses after sugammadex administration showed a temporary decrease after its initial recovery. Maximum decreases in twitch responses were observed 17 min after initial dose of sugammadex. Twitch responses recovered to their control values after additional doses of 3.5 mg/kg sugammadex (4 mg/kg in total). Time from sugammadex administration to maximum decreases in twitch responses is earlier than has been reported in adults (20-70 min). It is demonstrated that following neuromuscular block reversal with insufficient dose of sugammadex, there is a possibility of the recurrence of residual paralysis within less than 20 min in pediatric patients.


Developmental stages of Hepatozoon seurati (Laveran and Pettit 1911) comb. nov., a parasite of the corned viper Cerastes cerastes and the mosquito Culex pipiens from Egypt.
Developmental stages of Hepatozoon seurati (Laveran and Pettit 1911) comb. nov. are described from the tissues of the corned viper Cerastes cerastes, and from the vector Culex pipiens. The parasite described in the present study is firstly recorded as Haemogregarina seurati (Laveran and Pettit 1911) in the same host. After demonstration of the sporogonous development in the mosquito vector (C. pipiens) which showed all characteristics of the genus Hepatozoon (large oocysts containing many sporocysts producing numerous sporozoites), the parasite should be transferred into the genus Hepatozoon. The infected erythrocytes measured 20 ± 0.95 × 7.3 ± 0.85 μm; while uninfected cells measured 13.3 ± 1.04 × 7.5 ± 0.16 μm. Hypertrophy and faintly stained cytoplasm are mostly occurred in infected erythrocytes. Blood stages of the parasite were found exclusively in the erythrocytes in two forms: (1) small trophozoites (10.0 ± 0.52 × 3.0 ± 0.4 μm) and (2) long (mature) sausage-shaped (16.5 ± 1.5 × 3.5 ± 0.4 μm). Merogony occurred in the endothelial cells of the blood capillaries of lung, liver, and spleen. Mature meronts was 27.6 ± 0.7 × 17.5 ± 0.5 μm in diameter and contained 20-35 merozoites (averaged in 26). These merozoites measured 16.5 ± 1.5 × 3.5 ± 0.4 μm. Syzygy and gamogony occurred in the mosquito myxocoel till the 5th day post-infection (p.i.) while sporogony took place after 15 days p.i. On the third day p.i., a large spherical macrogamete of 29.0 ± 0.8 × 20.5 ± 0.6 μm containing a distinct nucleus in association with a single microgamete were observed. The microgamete was pyriform measured 8 ± 02 μm in length. It had a prominent nucleus and a long flagellum of at least 20.4 ± 1.3 μm in length. Fertilization occurred on the 3rd to the 4th days p.i. and the formed zygote developed into an oocyst in which repeated mitotic divisions with centripetal invaginations occurred producing sporoblasts. After sporulation, each sporoblast termed as sporocyst, and contained 18 banana-shaped sporozoites measured 14.0 ± 1.6 × 3.2 ± 0.6 μm. Experimental transmission was successful by intraperitoneal inoculation of the infective stages (sporozoites) to uninfected vipers and led to the appearance of blood stages after 5-6 weeks.


Association of p53 mutations with short survival in colorectal cancer.
Mutations in p53, a tumor suppressor gene located on chromosome 17p, are the most frequent genetic alterations found in human cancers. Increased intracellular concentration of p53, which is frequently but not systematically related to p53 mutation, has been proposed to be associated with poor prognosis in some tumor types. In colorectal cancer, this significance is still a matter of debate. To directly investigate the relationship between prognosis and p53 mutation, this study screened a series of 85 colorectal carcinomas for mutations in exons 5-8 of this gene. Polymerase chain reaction-amplified products from tumor DNA were analyzed by denaturing gradient gel electrophoresis and direct DNA sequencing. Forty-four tumors were found to be mutated (52%). A strong correlation between the presence of a mutation and short survival was observed (P = 0.003). When tumors were classified according to their histological stage, a multivariate Cox model analysis showed that p53 mutation, rather than 17p allelic loss (previously proposed to convey prognostic information), was retained as the only independent prognostic factor (relative risk, 2.25; 95% confidence interval, 1.06-4.80; P < 0.029). Combined with staging, direct monitoring of p53 mutation improves prognostic accuracy for colorectal cancer.


The effect of HIV-1 Gag myristoylation on membrane binding.
During the viral life cycle, an HIV protein, Gag, assembles at the host membrane, specifically at lipid raft regions, at very high concentrations leading to viral particle budding. Gag is post-translationally modified with an N-terminal myristate group which is thought to target Gag to lipid rafts thus aiding in assembly. Here we have analyzed the membrane binding of myristoylated HIV-1 Gag and a non-myristoylated form of HIV-1 Gag to various membrane models. After assessing the extent of myristoylation by HPLC and radiometric assays, we compared membrane binding using fluorescence methods. We found that myristoylated Gag shows a greater than twofold increase in binding affinity to model rafts. A structural model to explain these results is presented.


NAMPT (visfatin) and AKT1 genetic variants associate with myocardial infarction.
High plasma levels of the adipokine NAMPT (or visfatin) have been associated with cardiovascular disease. However experimental data suggest that NAMPT, via Akt signaling, protects the myocardium against hypoxic insults. We studied whether the NAMPT rs1319501, AKT1 rs3730358, p53 rs1042522, Mdm2 rs2279744 or eNOS rs1799983 SNP:s linked to NAMPT and Akt signaling associate with risk of myocardial infarction (MI). Cases were 828 men and 346 women aged 45-70 who had suffered a first MI. Control individuals, 1062 men and 513 women, were randomly chosen from the study base. We employed unconditional logistic regression analysis. The rs1319501 minor allele associated with MI among women aged 45-60; odds ratio (OR) under a recessive model of inheritance: 2.96 (95% confidence interval [CI] 1.06-8.29). Replication analysis in an independent material yielded OR point estimates in the same direction. The rs3730358 minor allele associated with low MI risk in men aged 45-60 (OR dominant model: 0.72, 95% CI 0.53-0.97), an association completely attenuated by adjusting for inflammatory markers. The NAMPT rs1319501 minor allele associates with increased MI risk in young women. In young men a protective effect of the AKT1 rs3730358 minor allele was suggested, possibly related to an attenuated inflammation.


The depolarization-induced outflow of D-[3H]aspartate from rat brain slices is modulated by metabotropic glutamate receptors.
Rat brain slices were used to study the effects of different metabotropic glutamate receptor ligands on (i) the depolarization (30 mM KCl)-induced outflow of previously taken up D-[3H]aspartate; (ii) the inhibition of forskolin (30 microM)-induced cyclic AMP accumulation; and (iii) the hydrolysis of phosphoinositides. In addition, the localization of mRNAs coding for different metabotropic glutamate receptor subtypes was detected using in situ hybridization. (1S-3R)-1-Aminocyclopentane-1,3-dicarboxylic acid (30-300 microM), a non selective metabotropic glutamate receptor agonist, significantly increased the KCl-induced output of radioactivity from cortical slices, whereas it inhibited the output from striatal slices. Conversely, (1S,3S,4S)-carboxycyclopropylglycine (0.1-1 microM), a relatively selective agonist of the mGluR2 metabotropic glutamate receptor subtype, had an inhibitory effect on the output of D-[3H]aspartate from both cortical and striatal slices and proved to be the most potent metabotropic glutamate receptor agonist in inhibiting cyclic AMP accumulation, but not in stimulating phosphoinositide hydrolysis. Since 2-amino-4-phosphonobutyrate (a mGluR4, mGluR6 and mGluR7 agonist) was not active in any of the assays tested, we hypothesized that the mGluR2 subtype could be involved in these events. Accordingly, mGluR2 mRNA expression was abundant in cortical neurons projecting to the striatum. Our experiments suggest that the stimulation of metabotropic glutamate receptors may either decrease or increase transmitter release depending on the subtype that prevails in the region under study.


Automobile air bags: friend or foe? A case of air bag-associated ocular trauma and a related literature review.
Although air bags are placed in automobiles to act as safety devices, they have been shown to carry a risk of injury themselves. Ocular injury, in particular, can often be a direct consequence of air bag deployment. A case of ocular air bag injury is presented. A discussion and review of the current literature on this issue follows. A 63-year-old man was transferred to our clinic after sustaining injuries related to a motor vehicle accident, during which the automobile's air bag was deployed. Initial examination revealed many signs of blunt ocular trauma of the O.D., including iridodialysis, dislocated lens with traumatic cataract, and traumatic/inflammatory glaucoma. Initial B-scan showed an attached retina O.D. One month later, the patient underwent an attempted pars plana vitrectomy with lensectomy, iris repair, and insertion of an anterior chamber intraocular lens. Complications arose during the procedure, and a total retinal detachment developed. Resultant acuity is no light perception O.D. Although ocular morbidity can be a direct consequence of air bag deployment, most eye injuries are minimal, and seem to be outweighed by the benefits of air bags. Drivers, as well as passengers, can minimize associated injuries by adhering to specific safety guidelines. This, as well as continual modification and improvement in air bag design, will maximize the safety of air bags and decrease the incidence of vision-threatening ocular injury caused by air bag deployment.


RNA-Based next generation sequencing complements but does not replace fluorescence in situ hybridization studies for the classification of aggressive B-Cell lymphomas.
Aggressive B-cell lymphomas are currently classified based in part upon the presence or absence of translocations involving BCL2, BCL6, and MYC. Most clinical laboratories employ fluorescence in situ hybridization (FISH) analysis for the detection of these rearrangements. The potential role of RNA-based sequencing approaches in the evaluation of malignant lymphoma is currently unclear. In this study, we performed RNA sequencing (RNAseq) in 37 cases of aggressive B-cell lymphomas using a commercially available next generation sequencing assay and compared results to previously performed FISH studies. RNAseq detected 1/7 MYC (14%), 3/8 BCL2 (38%) and 4/8 BCL6 (50%) translocations identified by FISH. RNAseq also detected 1 MYC/IGH fusion in a case not initially tested by FISH due to low MYC protein expression and 2 BCL6 translocations that were not detected by FISH. RNAseq identified the partner gene in each detected rearrangement, including a novel EIF4G1-BCL6 rearrangement. In summary, RNAseq complements FISH for the detection of rearrangements of BCL2, BCL6 and MYC in the evaluation and classification of aggressive B-cell lymphomas by detecting rearrangements that may be cryptic by FISH methods and by identifying the rearrangement partner genes. Detection of these clinically important translocations may be optimized by combined use of FISH and RNAseq.


Influenza presentations and use of neuraminidase inhibitors by Australian general practice registrars: a cross-sectional analysis from the ReCEnT study.
This study aims to establish prevalence and associations of (1) influenza and influenza-like illness (IILI) presentations to Australian general practice (GP) registrars (trainees) and (2) the use of neuraminidase inhibitors (NAIs) by GP registrars for new presentations of IILI, for the 10 years leading up to the COVID-19 pandemic in Australia (2010-2019). This was a cross-sectional analysis of the Registrar Clinical Encounters in Training ongoing inception cohort study of the in-consultation experience and clinical behaviours of GP registrars. Data are collected by individual registrars three times (from 60 consecutive consultations each time) at 6 monthly intervals. Data include diagnoses/problems managed and medicines prescribed, along with multiple other variables. Univariate and multivariable logistic regression was used to establish associations of registrars seeing patients with IILI and of prescribing NAIs for IILI. Teaching practices within the Australian general practitioner specialist vocational training programme. Practices were located in five of the six Australian states (plus one territory). GP registrars in each of their three compulsory 6-month GP training terms. From 2010 to 2019, 0.2% of diagnoses/problems seen by registrars were IILI. 15.4% of new IILI presentations were prescribed an NAI. IILI diagnoses were less likely in younger (0-14) and older (65+) age groups, and more likely in an area of higher socioeconomic advantage. There was considerable variation in NAI prescribing between regions. There was no significant association of prescribing NAIs with age or Aboriginal and/or Torres Strait Islander patients. IILI presentations were more likely among working-age adults and not among those groups at higher risk. Similarly, high-risk patient groups who would benefit most were not more likely to receive NAIs. The epidemiology and management of IILI has been distorted by the COVID-19 pandemic, but the burden of influenza in vulnerable populations must not be overlooked. Appropriately targeted antiviral therapy with NAIs influences outcomes for vulnerable patients. General practitioners manage the majority of IILI in Australia, and understanding GP IILI presentation and NAI prescribing patterns is a key first step to enabling sound and rational prescribing decisions for better patient outcomes.


Synthesis, antitumor activity and mechanism of action of novel 1,3-thiazole derivatives containing hydrazide-hydrazone and carboxamide moiety.
A series of novel 2,4,5-trisubstituted 1,3-thiazole derivatives containing hydrazide-hydrazine, and carboxamide moiety including 46 compounds T were synthesized, and evaluated for their antitumor activity in vitro against a panel of five human cancer cell lines. Eighteen title compounds T displayed higher inhibitory activity than that of 5-Fu against MCF-7, HepG2, BGC-823, Hela, and A549 cell lines. Especially, T1, T26 and T38 exhibit best cytotoxic activity with IC50 values of 2.21μg/mL, 1.67μg/mL and 1.11μg/mL, against MCF-7, BCG-823, and HepG2 cell lines, respectively. These results suggested that the combination of 1,3-thiazole, hydrazide-hydrazone, and carboxamide moiety was much favorable to cytotoxicity activity. Furthermore, the flow cytometry analysis revealed that compounds T1 and T38 could induce apoptosis in HepG2 cells, and it was confirmed T38 led the induction of cell apoptosis by S cell-cycle arrest.


[Our criteria for surgical treatment of acute obstructions of the large gut of neoplastic origin (author's transl)].
The authors review a total of 132 cases of acute neoplastic obstruction of the large gut seen in their clinical material over the last twenty years. In the light of the results obtained they discuss the indications and limitations of the various surgical strategies adopted (radical surgery, derivative surgery, and palliative resection); they conclude that radical procedures are best for the majority of cases.


Improving quality of life assessments for high-need adult Medicaid service users with mental health conditions.
There is a lack of consensus on how to evaluate health and social service programs for people with mental health (MH) conditions. Having service users be the primary decision makers in selecting outcome measures can inform a meaningful evaluation strategy. We sought to identify the quality of life (QoL) survey preferences of high-need adult service users with MH conditions. A systematic review identified generic, self-reported QoL surveys with evidence of validity in MH populations of interest. An advisory panel selected the most promising surveys to assess the success of programs like Medicaid for MH service users. Three groups of high-need, adult service users with MH conditions and one group of direct care staff ranked the surveys from the advisory panel, and generated and ranked characteristics that were desirable or undesirable in a QoL survey. Twenty-two surveys met the inclusion criteria. Of the six surveys selected by the advisory panel, groups of service users and direct care staff most preferred the Warwick-Edinburg Mental Well-being Scale (WEMWBS). The WEMWBS best embodied the features prioritized by the groups: to have a user-friendly format and positive focus, to be clearly worded and brief, and to avoid presumptive or unrealistic items. Service user groups appreciated survey topics most amenable to self-report, such as satisfaction with relationships. Using QoL surveys that service users prefer can reduce the chance that deteriorating QoL is going unchecked, and increase the chance that decisions based on survey findings are meaningful to service users.


Working status is related to post stroke/TIA cognitive decline: data from the TABASCO study.
Occupational status may influence physical and mental post-stroke outcomes. We aimed to evaluate the association between occupational status and type, or engagement in social and family activities, neuroimaging measures and cognitive decline (CD) in a prospective cohort of stroke patients. We included 273 first-ever stroke survivors at working age. All patients underwent 3T MRI at admission, as well as clinical and cognitive assessments at admission, 6, 12 and 24 months thereafter. Ninty nine (36.3%) of the participants were unemployed prior to the stroke. Age, sex, work type, other comorbidities, stroke severity or location were not associated with return to work. Patients who returned to work (87.4%) had better cognitive results and less depressive symptoms than those who retired after the event. Pre-stroke unemployment was associated with diabetes mellitus, hypertension, dyslipidemia, depression, poorer cognitive scores and brain atrophy. During the follow-up, 11% developed CD. CD was more common among previously unemployed than employed participants (19.2% vs. 6.3%, p = 0.001). Multiple regression adjusted for risk factors, revealed that pre-stroke unemployment was an independent predictor of CD (HR, 3.0; 95% CI: 1.06-8.44). Furthermore, engagement in mentally stimulating jobs decreased the risk for CD. Pre-stroke unemployment and post-stroke work disruption were each associated with depression and poorer cognitive performance up to two years post-stroke, as well as with brain atrophy at admission. Retirement after the stroke may increase the risk of developing CD. These results highlight the importance of continued employment in preserving cognitive abilities among stroke survivors.


Clinical scale isolation of T cell-depleted CD56+ donor lymphocytes in children.
We present a clinical scale method for immunomagnetic separation of CD56+ donor natural killer cells for adoptive immunotherapy of pediatric leukemias after allogeneic transplantation. This time-saving and partially automated procedure employed CD56+ selection followed by CD3+ depletion, resulting in a median purity of 98.6% NK cells and a four-log depletion of T cells. The enriched NK cells demonstrated high cytotoxic activity against K562 target cells and fresh leukemic blasts with low HLA class I expression, which could be further enhanced by IL-2 stimulation. Lysis of NK-insensitive leukemic cells with high HLA class I expression could also be demonstrated via ADCC. Due to the high degree of T cell depletion, alloreactive proliferation in mixed lymphocyte cultures and response to T cell-specific mitogen stimulation was profoundly decreased. Our results suggest that, even in the case of mismatched donors, infusions of donor NK cells with extremely low T cell content may be a promising treatment option for leukemic minimal residual disease after allogeneic transplantation without risk of inducing severe GVHD.


TFT sensor array for real-time cellular characterization, stimulation, impedance measurement and optical imaging of in-vitro neural cells.
Real-time in-vitro multi-modality characterization of neuronal cell ensemble involves highly complex interdependent phenomena and processes. Although a variety of microelectrode arrays (MEAs) have been reported, diagnosis techniques are limited in term of sensing area, optical transparency, resolution and number of modalities. This paper presents an optically transparent thin-film-transistor (TFT) array biosensor chip for neuronal ensemble investigation, in which TFT electrodes are used for six modalities including extracellular voltage recording of both action potential (AP) and local field potential (LFP), current or voltage stimulation, chemical stimulation, electrical impedance measurement, and optical imaging. The sensor incorporates a large sensing area (15.6 mm × 15.6 mm) with a 200 × 150 array of indium-tin-oxide (ITO) electrodes placed at a 50 μm or 100 μm pixel pitch and with 10 ms temporal resolution; these performances are comparable to the state-of-the-art MEA devices. The TFT electrode array is designed based on the switch matrix architecture. The reliability and stability of TFTs are examined by measuring their electrical characteristics. Impedance spectroscopy function is verified by mapping the neuron position and the status (cells alive or dead, contamination) on the electrodes, which facilitates the biochemical studies in electrical domain that adds quantitative views to visual observation of cells through the optical microscopy. An in-vitro neuron culture is studied using electrophysiological, electrochemical, and optical characterization. Detailed signal analysis is demonstrated to prove the capability of bioassay.


N-[11C]methylpiperidine esters as acetylcholinesterase substrates: an in vivo structure-reactivity study.
A series of simple esters incorporating the N-[11C]methylpiperidine structure were examined as in vivo substrates for acetylcholinesterase in mouse brain. 4-N-[11C]Methylpiperidinyl esters, including the acetate, propionate and isobutyrate esters, are good in vivo substrates for mammalian cholinesterases. Introduction of a methyl group at the 4-position of the 4-piperidinol esters, to form the ester of a teritary alcohol, effectively blocks enzymatic action. Methylation of 4-N-[11C]methylpiperidinyl propionate at the 3-position gives a derivative with increased in vivo reactivity toward acetylcholinesterase. Esters of piperidinecarboxylic acids (nipecotic, isonipecotic and pipecolinic acid ethyl esters) are not hydrolyzed by acetylcholinesterase in vivo, nor do they act as in vivo inhibitors of the enzyme. This study has identified simple methods to both increase and decrease the in vivo reactivity of piperidinyl esters toward acetylcholinesterase.


Diagnostic Workup for Patients with Solid Renal Masses: A Cost-Effectiveness Analysis.
For patients with solid renal masses, a precise differentiation between malignant and benign tumors is crucial for forward treatment management. Even though MRI and CT are often deemed as the gold standard in the diagnosis of solid renal masses, CEUS may also offer very high sensitivity in detection. The aim of this study therefore was to evaluate the effectiveness of CEUS from an economical point of view. A decision-making model based on a Markov model assessed expenses and utilities (in QALYs) associated with CEUS, MRI and CT. The utilized parameters were acquired from published research. Further, a Monte Carlo simulation-based deterministic sensitivity analysis of utilized variables with 30,000 repetitions was executed. The willingness-to-pay (WTP) is at USD 100,000/QALY. In the baseline, CT caused overall expenses of USD 10,285.58 and an efficacy of 11.95 QALYs, whereas MRI caused overall expenses of USD 7407.70 and an efficacy of 12.25. Further, CEUS caused overall expenses of USD 5539.78, with an efficacy of 12.44. Consequently, CT and MRI were dominated by CEUS, and CEUS remained cost-effective in the sensitivity analyses. CEUS should be considered as a cost-effective imaging strategy for the initial diagnostic workup and assessment of solid renal masses compared to CT and MRI.


Brain insulin signaling as a potential mediator of early life adversity effects on physical and mental health.
In numerous brain structures, insulin signaling modulates the homeostatic processes, sensitivity to reward pathways, executive function, memory, and cognition. Through human studies and animal models, mounting evidence implicates central insulin signaling in the metabolic, physiological, and psychological consequences of early life adversity. In this review, we describe the consequences of early life adversity in the brain where insulin signaling is a key factor and how insulin may moderate the effects of adversity on psychiatric and cardio-metabolic health outcomes. Further understanding of how early life adversity and insulin signaling impact specific brain regions and mental and physical health outcomes will assist in prevention, diagnosis, and potential intervention following early life adversity.


Immunotherapy for osteosarcoma: Fundamental mechanism, rationale, and recent breakthroughs.
Osteosarcoma (OS) is the most common primary malignancy of the bone and has a high propensity for local invasion and metastasis. Although combining surgery with chemotherapy has immensely improved the outcomes of osteosarcoma patients, the prognosis of metastatic or recurrent osteosarcomas is still unsatisfactory. Immunotherapy has proven to be a promising therapeutic strategy against human malignancies and improved understanding of the immune response to OS, and biomarker development has increased the number of patients who benefit from immunotherapies in recent years. Here, we review recent advances in immunotherapy in osteosarcoma and discuss the mechanisms and status of immunotherapies in both preclinical and clinical trials as well as future therapies on the horizon. These advances may pave the way for novel treatments requisite for patients with osteosarcoma in need of new therapies.


Usefulness and tolerability of hirulog, a direct thrombin-inhibitor, in unstable angina pectoris.
In an open-label pilot study of 20 patients with unstable angina (Braunwald class I-IIIB), hirulog was administered as a continuous intravenous infusion for 5 days in a dose of 0.2 mg/kg/hour to produce an activated partial thromboplastin time of approximately 200% of control. The primary end points of the study were: death, development of a transmural myocardial infarction, and intractable angina needing interventions such as an intraaortic balloon pump insertion, angioplasty and surgery. The secondary end points were the presence of an intracoronary thrombus detected on angiography and hemorrhagic complications during therapy. There was no death or transmural infarction in this study cohort; however, 1 patient developed intractable angina. Intracoronary thrombus was documented in 2 patients. Infusion of hirulog resulted in a steady prolongation of the activated partial thromboplastin time without any hemorrhagic or other adverse effect. Hirulog appears to be an effective antithrombotic agent that is tolerated well and may have advantages over heparin in the management of patients with unstable angina.


Digitalis and the autonomic nervous system.
Digitalis produces many of its effects in intact animals and human beings by modifying the properties of the autonomic nervous system. The parasympathetic limb of the autonomic nervous system is most sensitive to these effects of digitalis, and its properties are significantly altered with therapeutic concentrations of the drug. These actions are particularly important in mediating the electrophysiologic effects of digitalis. With toxic concentrations of digitalis, stimulation of sympathetic nerve activity may also occur. This latter action may be involved in the arrhythmogenic effects of digitalis. These effects of digitalis on the autonomic nervous system play a major role in determining the pharmacodynamic actions of the drug in patients. The effects of digitalis on the autonomic nervous system also provide a setting for important interactions with other drugs that modify the properties of the sympathetic and parasympathetic nervous systems.


Silencing of ALOX15 reduces ferroptosis and inflammation induced by cerebral ischemia-reperfusion by regulating PHD2/HIF2α signaling pathway.
To investigate the potential mechanism of arachidonic acid deoxyribozyme 15 (ALOX15) in ferroptosis and inflammation induced by cerebral ischemia reperfusion injury. The mice and cell models of cerebral ischemia-reperfusion injury were constructed. Western Blot was used to detect the protein expression levels of ALOX15, glutathione peroxidase (GPX4), hypoxia-inducible factor-2α (HIF-2α), prolyl hydroxylase (PHD) and inflammatory factors (NLRP3, IL-1β, IL-18) in brain tissues and cells. Cell proliferation activity was detected by CCK-8 method. LDH assay was used to detect the release of lactate dehydrogenase. TTC staining was used to observe cerebral infarction. In cerebral ischemia-reperfusion mice and cell models, the expression of ALOX15 protein was increased, the expression of GPX4, a key marker of ferroptosis was decreased, and silencing of ALOX15 down-regulated the GPX4 expression. HIF-2α expression was down-regulated in animal and cell models of cerebral ischemia reperfusion, and silencing of ALOX15 increased the HIF-2α expression by inhibiting PHD2 expression. Inhibition of ALOX15 expression reduced inflammatory factors levels (NLRP3, IL-1β, and IL-18) in cerebral ischemia. Inhibitor of PHD2 (IXOC-4) alleviating brain injury and cell death induced by cerebral ischemia reperfusion and stabilize HIF-2α expression in vivo. The expression of ALOX15 was up-regulated in cerebral ischemia-reperfusion animals and cells model. Inhibition of ALOX15 up-regulated the GPX4 expression, and promoted HIF-2α expression by inhibiting PHD2, thus alleviating ferroptosis and inflammation caused by cerebral ischemia-reperfusion injury.


Clinically meaningful efficacy of a non-estrogen agent: re-analysis of phase-2 data evaluating MF101.
To evaluate the level of reduction in hot flushes among a cohort of postmenopausal women randomized to a phase-2 clinical trial evaluating MF101, a selective estrogen receptor β modulator, for treatment of vasomotor symptoms to determine clinically meaningful efficacy. We performed a re-analysis of data from a double-blinded, placebo-controlled trial of 217 postmenopausal women randomized to treatment with MF101 or placebo for 12 weeks. At study termination, participants were asked if they were satisfied enough with medication to continue therapy. Of the women treated, 73% with ≥50% reduction in hot flush frequency and 77% with ≥60% reduction in hot flush frequency were willing to continue treatment. A 50-60% reduction in hot flushes is clinically meaningful among postmenopausal women who are being treated with a non-estrogen agent such as MF101.


Factors affecting the mortality of pediatric fulminant hepatic failure in relation to hepatitis B virus infection.
To investigate the factors affecting the outcome of fulminant hepatic failure (FHF) in children in relation to hepatitis B virus (HBV) infection. Retrospective review of a total of 94 cases (61 males and 33 females, aged from 1 month to 15 years) recruited from nine tertiary referral centers in Taiwan from 1985 to 1999. The overall mortality rate was 75%. Patients in the mortality group were of an older age, had higher peak total bilirubin levels, a longer prothrombin time, and a lower percentage of HBV positivity (P < 0.001, P = 0.003, P = 0.0027 and P = 0.042, respectively). Mortality was 65% in the HBV positive (n = 42) and 83% in the HBV negative (n = 52) group (P = 0.05). In the HBV positive group, the prothrombin time was noted to be the single factor affecting outcome (P = 0.036). In the HBV negative group, older age and higher peak value of total serum bilirubin were suggestive of poor survival rate (P < 0.001 and P = 0.006, respectively). Multivariate analysis revealed that total bilirubin was the single factor affecting outcome in the HBV-negative group. The mortality rate of HBV positive children in three consecutive time periods without liver transplantation (1985-1989, 1990-1994, 1995-1999) decreased gradually (91, 67 and 38%, respectively, with P = 0.027). This change was not observed in HBV-negative cases. Hepatitis B virus positive FHF had a lower mortality rate than HBV negative FHF, with each group having different factors affecting mortality.


c-fos and c-jun mRNA expression in a pig liver model of ischemia/reperfusion: effect of extended cold storage and the antioxidant idebenone.
Expression of immediate early genes has been reported during reperfusion after ischemia in rat livers due to oxygen radical formation. This study investigates in perfused pig livers the effect of the antioxidant idebenone and of cold ischemia time on the gene expression of c-fos and c-jun. Livers were perfused for 210 min after 0.5 h or 20 h ischemic storage (4 degrees C). One group of pigs was fed idebenone (280 mg/day/7days) prior to organ harvesting. C-fos and c-jun mRNA were determined by RT-PCR at 3, 30, 60, 120 180, 210 min during reperfusion. Lipid peroxidation increased in liver tissue form 0.54 +/- 0.21 to 1. 09 +/- 0.54 nmol MDA/mg protein during reperfusion after 20 h compared to 0.5 h cold storage. This was antagonized by idebenone (0. 68 +/- 0.20 nmol/MDA/mg protein). C-fos and c-jun were strongly induced in livers stored for 20 h, which was attenuated by idebenone (p < 0.05). These findings suggest that cold ischemia time and oxygen radicals are critical for immediate early gene expression and that application of an effective antioxidant can attenuate this early stress reaction of the pig liver.


Cell membrane destabilizes progressively during repetitive mechanical rupture events.
The postfusion oscillation cycle method of electrofused cells was applied to red blood cell membranes to induce repetitive membrane ruptures and test the mechanical membrane resistance against sequential events of membrane strain and rupture. After producing doublets from pairs of electrofused cells, they entered the oscillation cycle, providing a sequence of at least four consecutive colloidosmotic-driven rupture events. Different gradations of colloidosmotic pressure loads between 3230 Pa and 8640 Pa were established with various buffer types. The independence of buffer type and geometrical and mechanical observations has been verified independently for both parts of the oscillation sequence. With decreasing colloidosmotic inducement, caused by repetitive oscillation cycles, an increasing susceptibility of the cell membrane against membrane rupture was measurable. Since side-effects had been eliminated, it could be concluded that the cell membrane resistance against repetitive mechanical ruptures decreases.


Fast acrosome reaction measure: a highly sensitive method for evaluating stimulus-induced acrosome reaction.
To test a new method for evaluation of stimulus-induced acrosome reaction (AR). To determine whether this method, based on the definition of a specific staining pattern of recently reacted spermatozoa, brings an advantage of increased sensitivity as compared with a standard procedure. The hypothesis that a specific staining pattern with fluorescein-labeled Pisum sativum agglutinin corresponds to recently acrosome-reacted spermatozoa was tested by analyzing the frequency of this pattern at different time points after addition of ionophore A23187, an artificial AR inducer. The AR results obtained with a new method based on this relationship were compared with those obtained with a standard method of AR evaluation. Private hospital, public research center, and a university-based research laboratory. Healthy sperm donors. Changes in the frequency of specific sperm-staining patterns. A specific sperm-staining pattern with P. sativum agglutinin was shown to be associated with a recently occurred AR, whereas the absence of staining was typical of reactions having occurred a longer time ago. This phenomenon was used to define a formula for fast AR measure after stimulus addition. An increased sensitivity provided by this formula as compared with a standard evaluation was demonstrated. Fast AR measure is a simple, easy-to-perform method for AR evaluation. It is particularly suitable for testing the effects of rapidly acting stimuli.


Tumor vasculature normalization by orally fed erlotinib to modulate the tumor microenvironment for enhanced cancer nanomedicine and immunotherapy.
The abnormal tumor vasculature is one of key reasons that lead to the limited tumor perfusion as well as hypoxic and immunosuppressive tumor microenvironment (TME). Herein, we uncover that by normalizing the tumor vasculature with erlotinib, a specific inhibitor of epidermal growth factor receptor (EGFR), the tumor perfusion and tumor oxygenation statuses in different types of tumors including murine breast tumors, colorectal tumors, and squamous cell carcinoma tumors, could be remarkably enhanced. As the results, the tumor uptake of drug-loaded nanoparticles as well as their interstitial penetration within the tumor would be greatly increased for mice pre-treated with erlotinib at the oral feeding dose of 50 mg/kg, leading to remarkably improved chemotherapeutic efficacy of nanomedicine. On the other hand, owing to the erlotinib-induced normalization of tumor vasculatures, the relieved hypoxic state in the three different types of tumors could alter the immunosuppressive TME into immunosupportive. Such an effect together with the increased tumor retention of anti-PDL1 antibody, a clinically approved checkpoint blockade agent, finally contributes to the greatly improved tumor inhibition effect in cancer immunotherapy. Therefore, our work presents a general yet effective strategy using a clinical drug to enhance the efficacies of cancer nanomedicine and immunotherapy by normalizing tumor vasculatures and modulating TME.


Cystoscopic ureteral stent placement: techniques and tips.
We present a video demonstrating technical considerations and tips for cystoscopic placement of external, lighted, and internal ureteral stents. Cystoscopic ureteral stent placement is useful in cases where difficult pelvic periureter dissection is expected or encountered. In this video, we review cystoscopy basics, our approach to various types of retrograde stent placement, and performing retrograde pyelograms. Traditional external ureteral stent and lighted stent placement for prophylactic purposes are discussed, with attention to understanding stent markings, appropriate resistance, and steps for externalization. Internal, double-J ureteral stent placement with the use of fluoroscopy is initiated with placement of a guidewire. An open-ended ureteral catheter is advanced over the wire in the pelvic portion of the ureter, and a retrograde pyelogram is performed. The wire is reintroduced and the stent advanced to the renal pelvis under fluoroscopy. The proximal curl is confirmed to be in the appropriate position with fluoroscopy. The string attached to the stent is then cut and removed, the guidewire is removed, and the stent is deployed with the distal curl in the bladder. This video reviews key steps for cystoscopic ureteral stent placement in a prophylactic setting, cases of challenging anatomy, or ureteral injury.


Endorphins in psychiatry: an overview and a hypothesis.
This article presents an overview of the biochemistry, pharmacology, and physiology of endogenous opioid peptides (endorphins). Clinical psychopharmacology of exogenous opiate agonists and antagonists is reviewed. The evidence presented in the review is compatible with a hypothesis that the level of functional endorphins may be related to psychological events, with a normal level needed for psychological homeostasis. One corollary of this hypothesis is that the level of opioids in the brains of the mentally ill may be disturbed. Therapeutic implications of this hypothesis are considered.


Clinical utility of HPV-DNA detection: triage of minor cervical lesions, follow-up of women treated for high-grade CIN: an update of pooled evidence.
Human papilloma virus (HPV) testing and repeat cytology are both proposed as methods to triage women with minor cytological cervical lesions. By triage, those women can be identified who need referral for diagnostic exploration with colposcopy and/or biopsy. We conducted meta-analyses of reported studies on the accuracy to detect high-grade cervical intra-epithelial neoplasia or worse disease (CIN2+) in women with ASCUS or LSIL. We also performed meta-analyses to examine the best predictor of recurrence of CIN after treatment for CIN2 or CIN3. We found that HPV testing using the Hybrid Capture II test is more effective (more sensitive, equally specific) than cytology for the triage of patients with ASCUS Pap smears. Because of the high rate of HPV positivity, this is not the case for patients with LSIL. Studies concerning post-treatment follow-up were heterogeneous. In general, HPV testing performed better than follow-up cytology to predict success or failure of treatment (significantly higher sensitivity, not significantly lower specificity). Overall, in comparison with follow-up cytology, HPV DNA testing is more sensitive and equally specific for triage of ASCUS cases and for predicting recurrence of CIN in women treated for high-grade CIN.