Impact of heavy resistance training on pregnancy and postpartum health outcomes.
Participation in Olympic weightlifting, the Valsalva maneuver, and acute or prolonged supine weightlifting during pregnancy are cautioned against; however, these recommendations are based on expert opinions as opposed to empirical evidence. The aim of this study was to examine the training and health outcomes of individuals who engaged in heavy resistance training during pregnancy. A total of 679 individuals who lifted at least 80% one-repetition maximum during pregnancy participated in an online survey. Participants were primarily recreational athletes (88%, 332 out of 675) engaged in CrossFit™ (61%, 410 out of 675) and/or weightlifting (49%, 332 out of 675) during pregnancy. Most participants reported no complications during pregnancy or delivery (66%, 388 out of 589), whereas 57% (241 out of 426) reported urinary incontinence following pregnancy. Participants who maintained pre-pregnancy training levels until delivery reported significantly less reproductive complications than those who ceased training levels prior to delivery (p = 0.006). Most respondents engaged in Olympic lifting (72%, 311 out of 432) and lifting in a supine position (71%, 306 out of 437), whereas fewer reported use of the Valsalva maneuver during pregnancy (34%, 142 out of 412). Most returned to weightlifting following delivery (89%, 400 out of 447, average: 3.2 ± 3.0 months), including Olympic lifting (81%, 300 out of 372, average: 4.0 ± 3.4 months) and Valsalva (62%, 213 out of 341, average: 4.5 ± 3.6 months). Individuals who engaged in heavy prenatal resistance training had typical perinatal and pelvic floor health outcomes that were not altered whether they engaged in, or avoided Olympic lifting, Valsalva or supine weightlifting.


Use of RNA-seq data to identify and validate RT-qPCR reference genes for studying the tomato-Pseudomonas pathosystem.
The agronomical relevant tomato-Pseudomonas syringae pv. tomato pathosystem is widely used to explore and understand the underlying mechanisms of the plant immune response. Transcript abundance estimation, mainly through reverse transcription-quantitative PCR (RT-qPCR), is a common approach employed to investigate the possible role of a candidate gene in certain biological process under study. The accuracy of this technique relies heavily on the selection of adequate reference genes. Initially, genes derived from other techniques (such as Northern blots) were used as reference genes in RT-qPCR experiments, but recent studies in different systems suggest that many of these genes are not stably expressed. The development of high throughput transcriptomic techniques, such as RNA-seq, provides an opportunity for the identification of transcriptionally stable genes that can be adopted as novel and robust reference genes. Here we take advantage of a large set of RNA-seq data originating from tomato leaves infiltrated with different immunity inducers and bacterial strains. We assessed and validated 9 genes that are much more stable than two traditional reference genes. Specifically, ARD2 and VIN3 were the most stably expressed genes and consequently we propose they be adopted for RT-qPCR experiments involving this pathosystem.


Preparation of Fragaceatoxin C (FraC) Nanopores.
Biological nanopores are an emerging class of biosensors with high-end precision owing to their reproducible fabrication at the nanometer scale. Most notably, nanopore-based DNA sequencing applications are currently being commercialized, while nanopore-based proteomics may become a reality in the near future.Although membrane proteins often prove to be difficult to purify, we describe a straightforward protocol for the preparation of Fragaceatoxin C (FraC) nanopores, which may have applications for DNA analysis and nanopore-based proteomics. Recombinantly expressed FraC nanopores are purified via two rounds of Ni-NTA affinity chromatography before and after oligomerization on sphingomyelin-containing liposomes. Starting from a plasmid vector containing the FraC gene, our method allows the production of purified nanopores within a week. Afterward, the FraC nanopores can be stored at +4 °C for several months, or frozen.


Synchrotron X-ray microtransections: a non invasive approach for epileptic seizures arising from eloquent cortical areas.
Synchrotron-generated X-ray (SRX) microbeams deposit high radiation doses to submillimetric targets whilst minimizing irradiation of neighboring healthy tissue. We developed a new radiosurgical method which demonstrably transects cortical brain tissue without affecting adjacent regions. We made such image-guided SRX microtransections in the left somatosensory cortex in a rat model of generalized epilepsy using high radiation doses (820 Gy) in thin (200 μm) parallel slices of tissue. This procedure, targeting the brain volume from which seizures arose, altered the abnormal neuronal activities for at least 9 weeks, as evidenced by a decrease of seizure power and coherence between tissue slices in comparison to the contralateral cortex. The brain tissue located between transections stayed histologically normal, while the irradiated micro-slices remained devoid of myelin and neurons two months after irradiation. This pre-clinical proof of concept highlights the translational potential of non-invasive SRX transections for treating epilepsies that are not eligible for resective surgery.


Risk and outcomes of coronavirus disease in patients with inflammatory bowel disease: A systematic review and meta-analysis.
The risk of severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) infection and clinical outcomes of coronavirus disease (COVID-19) in inflammatory bowel disease are unclear. We searched PubMed and Embase with the keywords: inflammatory bowel disease, Crohn's disease, ulcerative colitis and COVID-19, novel coronavirus and SARS-CoV-2. We included studies reporting the frequency of COVID-19 infection and outcomes (hospitalisation, need for intensive care unit care and mortality) in patients with inflammatory bowel disease. We estimated the pooled incidence of COVID-19 in inflammatory bowel disease and comparative risk vis-a-vis the general population. We also estimated the pooled frequency of outcomes and compared them in patients who received and did not receive drugs for inflammatory bowel disease. Twenty-four studies were included. The pooled incidence rate of COVID-19 per 1000 patients of inflammatory bowel disease and the general population were 4.02 (95% confidence interval [CI, 1.44-11.17]) and 6.59 [3.25-13.35], respectively, with no increase in relative risk (0.47, 0.18-1.26) in inflammatory bowel disease. The relative risk of the acquisition of COVID-19 was not different between ulcerative colitis and Crohn's disease (1.03, 0.62-1.71). The pooled proportion of COVID-19-positive inflammatory bowel disease patients requiring hospitalisation and intensive care unit care was 27.29% and 5.33% while pooled mortality was 4.27%. The risk of adverse outcomes was higher in ulcerative colitis compared to Crohn's disease. The relative risks of hospitalisation, intensive care unit admission and mortality were lower for patients on biological agents (0.34, 0.19-0.61; 0.49, 0.33-0.72 and 0.22, 0.13-0.38, respectively) but higher with steroids (1.99, 1.64-2.40; 3.41, 2.28-5.11 and 2.70, 1.61-4.55) or 5-aminosalicylate (1.59, 1.39-1.82; 2.38, 1.26-4.48 and 2.62, 1.67-4.11) use. SARS-CoV-2 infection risk in patients with inflammatory bowel disease is comparable to the general population. Outcomes of COVID-19-positive inflammatory bowel disease patients are worse in ulcerative colitis, those on steroids or 5-aminosalicylates but outcomes are better with biological agents.


Tardive dysphoria: antidepressant-induced chronic depression.
A significant number of depressed individuals experience inadequate benefit from long-term antidepressant use. This paper investigates the hypothesis that in some individuals persistent use of antidepressants may be prodepressant. Literature regarding the effect of long-term use of antidepressants was reviewed by searching PubMed and Ovid data bases with terms: antidepressant tachyphylaxis, treatment-resistant depression, chronic depression and antidepressant tolerance. Antidepressant treatment-resistant patients frequently had a positive initial response to antidepressants. When resistance appeared, initial increases in dose or medication changes usually resulted in transient improvement. Eventually, the episodic course of the original depressive illness was replaced with a continuous, unresponsive depressive syndrome. We propose the term tardive dysphoria to describe this phenomenon. The phenomenon of antidepressantinduced depression, or tardive dysphoria, needs to be experimentally examined in blinded, randomised antidepressant discontinuation studies.


The use of dynamic positron emission tomography imaging for evaluating the carcinogenic progression of intestinal metaplasia to esophageal adenocarcinoma.
In this study, we investigate the use of PET scanning in the carcinogenic progression of reflux esophagitis to Barrett's esophagus to high grade dysplasia to esophageal adenocarcinoma, and correlate the uptake levels of 18F-FDG related to histological changes, and the rates of proliferation and apoptosis. An established esophagoduodenal anastomsis rat model in conjunction with micro-PET scanning at 1 week, 1 month, 3 month, and 6 month after procedure was performed. Increased uptake levels of 18F-FDG were observed in the esophagi after EDA procedure. The higher level of 18F-FDG uptake within esophageal epithelium was identified in intestinal metaplastic transformation and esophagoduodenal adenocarcinoma by histological examination. Dynamic PET scanning represents a powerful tool in analyzing morphological carcinogenic transformation non-invasively in the esophagus. 18F- FDG accumulation was a sensitive marker in reflux esophageal injury carcinogenic progression from intestinal metaplasia to EAC.


Orally active epoxyeicosatrienoic acid analogs in hypertension and renal injury.
Epoxyeicosatrienoic acids (EETs) are arachidonic acid metabolites synthesized by cytochrome P450 epoxygenases. Biological activities for EETs include vasodilation, decreasing inflammation, opposing apoptosis, and inhibiting renal sodium reabsorption. These actions are beneficial in lowering blood pressure and slowing kidney disease progression. Furthermore, evidence in human and experimental animal studies have found that decreased EET levels contribute to hypertension and kidney diseases. Consequently, EET mimics/analogs have been developed as a potential therapeutic for hypertension and acute and chronic kidney diseases. Their development has resulted in EET analogs that are orally active with favorable pharmacological profiles. Analogs for 8,9-EET, 11,12-EET, and 14,15-EET have been tested in several hypertension and kidney disease animal models. More recently, kidney targeted EET analogs have been synthesized and tested against drug-induced nephrotoxicity. Experimental evidence has demonstrated compelling therapeutic potential for EET analogs to oppose cardiovascular and kidney diseases. These EET analogs lower blood pressure, decrease kidney inflammation, improve vascular endothelial function, and decrease kidney fibrosis and apoptosis. Overall, these preclinical studies support the likelihood that EET analogs will advance to clinical trials for hypertension and associated comorbidities or acute and chronic kidney diseases.


A cohort investigation of the changes in vocational dental practitioners' views on repairing defective direct composite restorations.
To investigate changes in vocational dental practitioners (VDPs) views on repairing direct composite restorations (DCRs) during the vocational training (VT) year 2002-2003. A questionnaire-based cohort study. Data on the repair of DCRs, including indications and details of operative techniques were collected by a questionnaire sent at the beginning of the VT year (September 2002) to all 12 Postgraduate Dental Deans in England. A follow-up questionnaire was sent to the Deans near the end of the VT year (May 2003). They were asked, on both occasions, to distribute the questionnaires among their VDPs and to return the completed questionnaires. Ten deaneries participated in the study. The September 2002 and May 2003 findings (306 respondents [66%] and 313 respondents [67%] respectively) were analysed using Chi-square statistic and t-test to identify any significant differences (P<0.05). Significant changes in the attitudes of the VDPs during the VT year included a shift to viewing reduction in treatment time (30% to 48%), cost to patient (12% to 20%) and the management of the partial loss of a restoration (57% to 70%) as indications for repairs. In contrast, there was a significant change in viewing secondary caries as an indication for repair (46% to 29%). In addition, there was a significant, worrisome shift in relation to the use of bonding systems and finishing instruments, together with a reduction in the anticipated longevity of composite repairs. Aspects of VDPs' attitudes to the repair of DCRs were found to change significantly during the VT year. Not all aspects of the changes observed were considered to be positive. Knowledge and understanding of repairs to DCRs need to be strengthened.


Long-term developmental outcome of children with a fetal diagnosis of isolated inferior vermian hypoplasia.
Isolated inferior vermian hypoplasia (iiVH) is one of the most common fetal cerebellar anomalies presenting for fetal neurological counselling with controversial postnatal neurodevelopmental outcome. In the present study, we characterised the long-term neurodevelopmental outcome of prenatally diagnosed iiVH at school age. We prospectively followed 20 children with fetal MRI diagnosis of iiVH including their postnatal MRI result and developmental outcome at school age (mean 6.1 years±1.9 years SD) using a comprehensive age-appropriate developmental testing battery, which encompassed cognitive, language, social and behavioural domains. Parental stress level and socioeconomic status were also evaluated. All children with postnatally confirmed iiVH had a normal neurodevelopmental outcome. A subgroup of children (2/20) who demonstrated cognitive delays and behavioural impairments had more extensive cerebellar malformation. Despite a normal developmental outcome, the parents of children with postnatally confirmed iiVH had higher parental stress compared with those parents whose children had normal postnatal MRI. Children with postnatally confirmed iiVH show age appropriate functioning at school age. Postnatal MRI is important to confirm the diagnosis of iiVH and to exclude associated anomalies that impact neurodevelopmental outcome. A diagnosis of iiVH is associated with persistent elevated parental stress despite normal developmental outcomes in these children suggesting the need for ongoing parental support.


Osthole, a coumadin analog from Cnidium monnieri (L.) Cusson, stimulates corticosterone secretion by increasing steroidogenic enzyme expression in mouse Y1 adrenocortical tumor cells.
Osthole is an O-methylated coumadin, which was isolated and purified from the seeds of Cnidium monnieri (L.) Cusson. Osthole is a commonly used traditional Chinese medicine to treat patients with Kidney-Yang deficiency patients, who exhibit clinical signs similar to those of glucocorticoid withdrawal. However, the mechanism of action of osthole is not fully understood. This study was designed to reveal the effects of osthole on corticosterone production in mouse Y1 cell. Mouse Y1 adrenocortical cells were used to evaluate corticosterone production, which was quantified by enzyme-linked immunosorbent assay (ELISA) kits. Cell viability was tested using the MTT assay, and the mRNA and protein expression of genes encoding steroidogenic enzymes and transcription factors was monitored by quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) and western blotting, respectively. Osthole stimulated corticosterone secretion from mouse Y1 cells in a dose- and time-dependent manner, and osthole enhanced the effect of dibutyryl-cAMP (Bu2cAMP) on corticosterone production. Further, osthole also increased StAR and CYP11B1 mRNA expression in a dose-dependent manner and enhanced the expression of transcription factors such as HSD3B1, FDX1, POR and RXRα as well as immediate early genes such as NR4A1. Moreover, osthole significantly increased SCARB1(SRB1) mRNA and StAR protein expression in the presence or absence of Bu2cAMP; these proteins are an important for the transport of the corticosteroid precursor cholesterol transport into mitochondria. Our results show that the promotion of corticosterone biosynthesis and secretion is a novel effect of osthole, suggesting that this agent can be utilized for the prevention and treatment of Kidney-Yang deficiency syndrome.


Awareness of human papillomavirus and acceptability of the vaccine among women in Palestine: is it time for policy adjustment?
Progress has been made in the reduction of morbidity and mortality of cervical cancer by the implementation of human papillomavirus (HPV) vaccination programs. This study aimed to assess the awareness of Palestinian women about HPV as well as their knowledge and acceptability of the HPV vaccine and to examine the factors associated with good awareness. This was a national cross-sectional study. Adult women were recruited from hospitals, primary healthcare centers, and public spaces in 11 Palestinian governorates using convenience sampling. A structured questionnaire was used for data collection. For each correctly answered question, one point was given. The total score was calculated and categorized into poor (0-10) and good awareness (11-21). The questionnaire was completed by 7223 women out of 8086 who were approached (response rate = 89.3%). A total of 7058 questionnaires were included in the final analysis; 4403 from the West Bank and Jerusalem (WBJ) and 2655 from the Gaza Strip. Women recruited from the Gaza Strip were younger, getting lower monthly incomes, and with fewer chronic diseases than women recruited from the WBJ. Only 33 women (0.5%) displayed good awareness of HPV and its vaccine with 0.7% of women from WBJ and only 0.2% of women from the Gaza Strip. Completing post-secondary education, being employed or a student, and having a higher monthly income were associated with an increase in the likelihood of having good awareness. Among women who had heard of HPV (n = 571, 8.1%), only 46 women (8.1%) reported familiarity with its vaccine. Women from the WBJ were more likely than women from the Gaza Strip to have heard about the HPV vaccine (0.9% vs. 0.2%). Most women agreed to receive the HPV vaccine themselves or for their daughters if it was given without cost or with a co-payment. No differences were found in the likelihood of agreeing to receive the HPV vaccine among women in the WBJ versus the Gaza Strip. The overall awareness of HPV and its vaccine was extremely low. Inclusion of the HPV vaccine in the national immunization program could change this, especially as the HPV vaccine appeared to be acceptable.


Vascular Endothelial Growth Factor in Plasma and Pleural Effusion Is a Biomarker for Outcome After Bevacizumab plus Carboplatin-Paclitaxel Treatment for Non-small Cell Lung Cancer with Malignant Pleural Effusion.
Malignant effusion is associated with high serum and plasma levels of vascular endothelial growth factor (VEGF). There are no biomarkers of outcome for bevacizumab treatment in patients with malignant pleural effusion (MPE). We previously reported that carboplatin-paclitaxel plus bevacizumab was effective for patients with advanced non-squamous non-small cell lung cancer (NSCLC) and MPE, although we did not evaluate the relationship between treatment outcomes and plasma or pleural effusion levels of VEGF. Therefore, this study evaluated whether plasma or pleural effusion VEGF might predict bevacizumab treatment outcome. We enrolled 23 patients with NSCLC and MPE between September 2010 and June 2012. Plasma VEGF levels were measured in 19 patients and pleural VEGF levels were measured in 22 patients. Compared to patients with a low plasma VEGF level, patients with a high level exhibited significantly shorter overall survival (OS: 13.8 vs. 6.5 months, p=0.04), progression-free survival (PFS: 8.7 vs. 4.8 months, p<0.01), and period to re-accumulation of MPE (pPFS: 9.7 vs. 6.2 months, p=0.02). Compared to patients with a low VEGF level in pleural effusion, patients with a high VEGF level exhibited significantly shorter OS (19.6 vs. 6.9 months, p<0.01) and pPFS (9.6 vs. 6.7 months, p=0.04), although there was no significant difference in their PFS (6.6 vs. 5.9 months, p=0.18). VEGF levels in the plasma and pleural effusion may predict the outcome of bevacizumab treatment in patients with NSCLC and MPE.


A novel enhanced intensity-based automatic registration: Augmented reality for visualization and localization cancer tumors.
The purpose of this study is to replace the manual process (selecting the landmarks on mesh and anchor points on the video) by Intensity-based Automatic Registration method to reach registration accuracy and low processing time. The proposed system consists of an Enhanced Intensity-based Automatic Registration (EIbAR) using Modified Zero Normalized Cross Correlation (MZNCC) algorithm. The proposed system was implemented on videos of breast cancer tumors. Results showed that the proposed algorithm-as compared to a reference-improved registration accuracy by an average of 2 mm. In addition, the proposed algorithm-as compared to a reference-reduced the number of pixel matching, thereby reducing processing time on the video by an average of 22 ms/frame. The proposed system can, thus, provide an acceptable accuracy and processing time during scene augmentation of videos, which provides a seamless use of augmented-reality for surgeons in visualizing cancer tumors.


Combining individual and aggregated data to investigate the role of socioeconomic disparities on cancer burden in Italy.
Quantifying socioeconomic disparities and understanding the roots of inequalities are growing topics in cancer research. However, socioeconomic differences are challenging to investigate mainly due to the lack of accurate data at individual-level, while aggregate indicators are only partially informative. We implemented a multiple imputation algorithm within a statistical matching framework that combines diverse sources of data to estimate individual-level associations between income and risk of breast and lung cancer, adjusting for potential confounding factors in Italy. The framework is computationally flexible and can be adapted to similar contexts.


Reporting the methodology of height and weight acquisition in studies of body mass index-based prognosis in critically ill patients.
Conflicting findings were reported on the body mass index (BMI)-based prognosis of critically patients. Errors in source weight and height data can confound BMI group allocation. The aim of the present work was to examine investigators' reporting on the methods of height and weight acquisition (HWA). PubMed and Embase databases were searched for studies describing BMI group-based risk of death in critically ill patients. Eligible studies were examined for reporting on (1) the use of measured and/or estimated HWA, (2) details of measuring devices, (3) device accuracy, and (4) methods of adjustment for acute and chronic fluid-related weight changes. Thirty studies met the eligibility criteria, including 159,565 patients. No data were provided in 13 studies (52% of reported patients) on whether estimates or measurements were used for HWA. Measured HWA was used exclusively in 6 studies (3% of patients), and an unspecified combination of estimated and/or measured HWA was reported for the remainder. Only 1 study reported the specific devices used. None of the studies provided data on the bias and precision of measuring devices. Adjustment for chronic and/or acute fluid-related weight changes was addressed in 2 studies for each. These findings demonstrate the prevalent risk for BMI group misallocation in the reviewed studies, which may confound BMI-based prognosis, raising concerns about the validity of reported BMI-related prognostic impact.


Neuroprotective effect of the new thiadiazolidinone NP00111 against oxygen-glucose deprivation in rat hippocampal slices: implication of ERK1/2 and PPARgamma receptors.
Thiadiazolidinones (TDZDs) are small molecules that inhibit glycogen synthase kinase 3-beta (GSK3-beta) activity in a non competitive manner to ATP. NP00111, a new TDZD, besides causing inhibition of GSK-3beta, has also shown to be an agonist of PPARgamma . Since phosphorylation and consequent inhibition of GSK-3beta by PI-3K/Akt and agonism of PPARgamma have shown to afford neuroprotection in several in vitro and in vivo models, we have studied the potential neuroprotective effect of NP00111 in an "in vitro" model of ischemia-reperfusion. NP00111, at the concentration of 10 microM, significantly protected adult rat hippocampal slices subjected to oxygen and glucose deprivation (OGD) for 1 h followed by 3 h re-oxygenation, measured as lactic dehydrogenase (LDH) released to the extracellular media. The protective effects of NP00111 were more pronounced during the re-oxygenation period in comparison to the OGD period. Other GSK-3beta inhibitors like lithium or AR-A014418 did not afford protection in this model. However, the PPARgamma agonist rosiglitazone was protective at 3 microM. Protection afforded by NP00111 and rosiglitazone were prevented by the PPARgamma antagonist GW9662, suggesting that both NP00111 and rosiglitazone were preventing cell death caused by oxygen-glucose deprivation via activation of PPARgamma. NP00111 increased by two fold phosphorylation of ERK1/2 and its protective effects were lost when the hippocampal slices were co-incubated with the mitogen-activated protein kinase (MAPK) inhibitor PD98059. In conclusion, the novel TDZD NP00111 was protective against OGD in rat hippocampal slices by a mechanism related to phosphorylation of ERK1/2 via activation of PPARgamma.


Generation and subcellular distribution of histamine in human blood monocytes and monocyte subsets.
This study was designed to establish the sites of formation and storage of histamine and histidine decarboxylase (HDC) in human monocytes and two of their subsets. The experiments were carried out using monocytes from buffy coats of healthy blood donors. Histamine was quantitated by RIA, HDC activity by the formation of histamine. The monocyte subtype RM3/1 contained significantly more histamine than the subset 27E10 (0.041+/-0.025 vs. 0.005+/-0.004 pg/cell, p < 0.05) and also more HDC activity and HDC mRNA. After fractionation of monocyte homogenates in a discontinuous Percoll gradient or by differential centrifugation more than 80% of both, HDC activity and histamine, were recovered from the cytosolic fractions. About 50% of this histamine was found to be bound to proteins. In monocytes histamine and HDC are colocalized in the cytoplasm indicating a subcellular distribution different from mast cells or basophils. The data also show that histamine is synthesized by the monocytes themselves.


Detection, management, and prevention of diabetes-related foot disease in the Australian context.
Diabetes-related foot disease (DFD) is a widely feared complication among people who live with diabetes. In Australia and globally, rates of disability, cardio-vascular disease, lower extremity amputation, and mortality are significantly increased in patients with DFD. In order to understand and prevent these outcomes, we analyse the common pathogenetic processes of neuropathy, arterial disease, and infection. The review then summarises important management considerations through the interdisciplinary lens. Using Australian and international guidelines, we offer a stepwise, evidence-based practical approach to the care of patients with DFD.


A non-linear mathematical model for the in vivo determination of Kupffer cells number and rate of phagocytosis of radiocolloids in rats.
In order to perform a quantification of the hepatic RES function, a mathematical model of the colloids phagocytosis was constructed and validated in normal and partially hepatectomised rats. The experimental design consisted of the collection of successive blood samples for the measurement of radiocolloids time courses after the injection of different doses of gelatin colloids (0.075--5 mg of gelatin/100 g body wt). The unknown parameters were estimated by method of maximum likelihood using a second order algorithm. A good fit between experimental and simulated data was obtained for a large range of injected doses using a single set of parameters. Comparisons of parameters values between normal and hepatectomised rats were found to be -onsistent with the hepatectomy ratio. This computerised estimation of parameters provides a determination of both the total number of Kupffer cells and the mean time of a complete phagocytosis cycle which cannot be obtained by classical approaches.


Resveratrol alleviates hypoxia/reoxygenation injury‑induced mitochondrial oxidative stress in cardiomyocytes.
Resveratrol (RES) is a naturally occurring antioxidant compound found in red wine. Although it has been demonstrated to have a cardioprotective effect, the mechanism underlying this effect remains to be fully elucidated. The aim of the present study was to determine whether RES exerts a protective effect against mitochondrial oxidative stress and apoptosis in neonatal rat cardiomyocytes (NRCMs) induced by hypoxia/reoxygenation (H/R) injury. Primary cultured NRCMs were used as a model system and were divided into four experimental groups: Control, H/R, H/R + DMSO (H/R with 0.2% DMSO) and H/R + RES (H/R with 100 µM RES) groups. Mitochondrial oxidative stress was determined by measuring the alteration in the mitochondrial membrane potential (ΔΨm) of NRCMs, the release of lactate dehydrogenase (LDH) and the ratio of B‑cell lymphoma 2 (Bcl‑2)/Bcl‑2‑associated X protein (Bax) from NRCMs. Cell apoptosis was assessed by measuring cell apoptotic rates and the activity of caspase 3. In the H/R+RES group, RES significantly alleviated structural impairment, including disordered α‑actin and F‑actin, in the NRCMs induced by H/R injury. RES attenuated H/R injury‑induced mitochondria oxidative stress. RES also attenuated H/R injury‑induced cell apoptosis; it decreased the NRCM apoptotic rate from 84.25±7.41% (H/R) to 46.39±5.43% (H/R+RES) (P<0.05, n=4), rescued the decrease in the Bcl2/Bax ratio induced by H/R from 0.53±0.08‑fold (H/R) to 0.86±0.06‑fold (H/R+RES) (P<0.05, n=5) and alleviated the increased activity of caspase 3 induced by H/R from 1.32±0.06‑fold to 1.02±0.04‑fold (P<0.05, n=5). Furthermore, RES significantly attenuated the increment of LDH release induced by H/R injury in NRCMs from 1.41±0.03‑fold (H/R) to 1.02±0.06‑fold (H/R+RES) (P<0.01, n=4) and alleviated the depolarization of ΔΨm induced by H/R, shifting the ratio of JC‑1 monomer from 62.39±1.82% (H/R) to 35.31±8.63% (H/R+RES) (P<0.05, n=4). RES alleviated the decrease in sirtuin 1 induced by H/R injury from 0.61±0.06‑fold (H/R) to 1.01±0.05‑fold (H/R+RES) (P<0.05, n=5). In conclusion, the present study is the first, to the best of our knowledge, to demonstrate that RES provides cardioprotection against H/R injury through decreasing mitochondria‑mediated oxidative stress injury and structural impairment in NRCMs. These results provide scientific evidence for the clinical application of RES in the treatment of cardiac conditions.


Sorafenib treatment in children with relapsed and refractory neuroblastoma: an experience of four cases.
Metastatic neuroblastoma is an aggressive malignancy with a poor prognosis. Recent findings have shown that sorafenib decreases cell viability and increases apoptosis in human neuroblastoma cell lines. We report an experience of compassionate use of sorafenib in children with treatment-refractory neuroblastoma. Sorafenib showed transient anti-tumor activity in all four patients without adverse effects. However, progression was observed after a short stabilization phase. While sorafenib showed minimal anti-tumor activity in our patients, it might still be effective in patients with neuroblastoma in an earlier stage.


General population screening for paediatric type 1 diabetes-A qualitative study of UK professional stakeholders.
Identifying children at risk of type 1 diabetes allows education for symptom recognition and monitoring to reduce the risk of diabetic ketoacidosis at presentation. We aimed to explore stakeholder views towards paediatric general population screening for type 1 diabetes in the United Kingdom (UK). Qualitative interviews were undertaken with 25 stakeholders, including diabetes specialists, policymakers and community stakeholders who could be involved in a future type 1 diabetes screening programme in the UK. A thematic framework analysis was performed using the National Screening Committee's evaluative criteria as the overarching framework. Diabetic ketoacidosis prevention was felt to be a priority and proposed benefits of screening included education, monitoring and helping the family to better prepare for a future with type 1 diabetes. However, diabetes specialists were cautious about general population screening because of lack of evidence for public acceptability. Concerns were raised about the harms of living with risk, provoking health anxiety and threatening the child's right to an 'open future'. Support systems that met the clinical and psychological needs of the family living with risk were considered essential. Stakeholders were supportive of research into general population screening and acknowledged this would be a priority if an immunoprevention agent were licensed in the UK. Although stakeholders suggested the harms of UK paediatric general population screening currently outweigh the benefits, this view would potentially be altered if prevention therapies were licensed. In this case, an evidence-based screening strategy would need to be formulated and public acceptability explored.


Synthesis of N-o-methylphenyl-N'-(sodium p-aminobenzene-sulfonate) thiourea and its chromogenic reaction with palladium(II).
A new chromogenic reagent, N-o-methylphenyl-N'-(sodium p-aminobenzenesulfonate)thiourea (MSAT), has been synthesized and characterized by elemental analysis, (1)H-NMR, FT-IR and UV-Vis spectra. Based on the absorption spectrum of the colored complex of MSAT with palladium(II), a novel spectrophotometric method for the determination of palladium has been developed. In a pH 4.0 - 5.5 HAc-NaAc buffer solution, palladium(II) reacted with MSAT to form a stable yellow water-soluble complex with an apparent molar absorptivity of epsilon = 2.04 x 10(5) L mol(-1) cm(-1) at the maximum absorption of 318.0 nm. Beer's law was obeyed in the concentration range of 1.2 - 11.8 microg per 25 mL for palladium(II) with a correlation coefficient of 0.9997. The probable interfering ions and their tolerable limits have also been investigated in detail. The proposed method is simple, rapid, and sensitive, and has been applied to the determination of palladium in anode mud and ore samples with satisfactory results.


Monitoring lower limb biomechanical asymmetry and psychological measures in athletic populations-A scoping review.
Lower limb biomechanics, including asymmetry, are frequently monitored to determine sport performance level and injury risk. However, contributing factors extend beyond biomechanical and asymmetry measures to include psychological, sociological, and environmental factors. Unfortunately, inadequate research has been conducted using holistic biopsychosocial models to characterize sport performance and injury risk. Therefore, this scoping review summarized the research landscape of studies concurrently assessing measures of lower limb biomechanics, asymmetry, and introspective psychological state (e.g., pain, fatigue, perceived exertion, stress, etc.) in healthy, competitive athletes. A systematic search of MEDLINE, Embase, CINAHL, SPORTDiscus, and Web of Science Core Collections was designed and conducted in accordance with PRISMA guidelines. Fifty-one articles were included in this review. Significant relationships between biomechanics (k = 22 studies) or asymmetry (k = 20 studies) and introspective state were found. Increased self-reported pain was associated with decreased range of motion, strength, and increased lower limb asymmetry. Higher ratings of perceived exertion were related to increased lower limb asymmetry, self-reported muscle soreness, and worse jump performance. Few studies (k = 4) monitored athletes longitudinally throughout one or more competitive season(s). This review highlights the need for concurrent analysis of introspective, psychological state, and biomechanical asymmetry measures along with longitudinal research to understand the contributing factors to sport performance and injury risk from biopsychosocial modeling. In doing so, this framework of biopsychosocial preventive and prognostic patient-centered practices may provide an actionable means of optimizing health, well-being, and sport performance in competitive athletes.


Delphinium uncinatum mediated biosynthesis of zinc oxide nanoparticles and in-vitro evaluation of their antioxidant, cytotoxic, antimicrobial, anti-diabetic, anti-inflammatory, and anti-aging activities.
Nanotechnology is perhaps the most widely explored scientific domain in the current era. With the advent of NPs, revolutionary changes have been observed in various scientific disciplines. Among the NPs, ZnO-NPs are the center of contemplation owing to their biocompatible nature. These nanoparticles have been prepared using a number of techniques; however, biological methods are among the most popular synthesis approaches. The current research therefore reports the phyto-fabrication of ZnO-NPs mediated by Delphinium uncinatum root extract. The resulting NPs were subjected to standard characterization methods such fourier transformed infrared spectroscopy, X-ray diffraction and transmission electron microscopy. The resulting NPs are exploited to their possible antioxidant, antimicrobial, antidiabetic, cytotoxic, anti-inflammatory and anti-ageing potency. FTIR confirmed the capping of ZnO-NPs by a variety of phytochemicals. ZnO-NPs average size was approximately 30 nm. ZnO-NPs exhibited substantial bio-potency and proved to be highly biocompatible even at higher concentrations. ZnO-NPs revealed strong antimicrobial potency for Pseudomonas aeruginosa proving to be the most susceptible strain showing inhibition of 16 ± 0.98. ZnO-NPs also showed dose dependent antidiabetic and cytotoxic potential. COX-1, COX-2, 15-LOX and sPLA2 were efficiently inhibited upon exposure to ZnO-NPs confirming the anti-inflammatory potential of ZnO-NPs. Similarly, ZnO-NPs also revealed considerable anti-aging potential. With such diverse biological potentials, ZnO-NPs can prove to be a potent weapon against a plethora of diseases; however, further study is necessary in order to discover the precise mechanism that is responsible for the biological potency of these NPs.


The relevance of subtyping children with mathematical learning disabilities.
Profiles of mathematical learning disability (MLD) have been conceptualized in the literature, but empirical evidence to support them based on academic and cognitive characteristics is lacking. We examined whether profiles of mathematics performance can empirically be identified and whether the identified profiles also differ in underlying cognitive skills. Latent profile analysis in 281 fourth-graders. Basic arithmetic and advanced mathematics were used to identify profiles. Cognitive skills were then described for each profile of mathematics performance. Four profiles of mathematics performance were retrieved from the data, including one general low-achieving profile. Additional profiles of MLD were not found, possibly because individual variation was substantial. It is highly important to understand children's mathematics performance from an individual perspective, rather than by averaging these children over subgroups. These new insights can be used to better tend to the specific needs of children with mathematical difficulties.


Disruption of the copper efflux pump (CopA) of Serratia marcescens ATCC 274 pleiotropically affects copper sensitivity and production of the tripyrrole secondary metabolite, prodigiosin.
The prodigiosin biosynthetic gene cluster (pig cluster) of Serratia marcescens ATCC 274 (Sma 274) is flanked by cueR/copA homologues. Inactivation of the copA homologue resulted in an increased sensitivity to copper, confirming that CopA is involved in copper homeostasis in Sma 274. The effect of copper on the biosynthesis of prodigiosin in Sma 274 and the copA mutant strain was investigated. Increased levels of copper were found to reduce prodigiosin production in the wild type Sma 274, but increase production in the copA mutant strain. The physiological implications for CopA mediated prodigiosin production are discussed. We also demonstrate that the gene products of pigB-pigE of Sma 274 are sufficient for the biosynthesis of 2-methyl-3-n-amyl-pyrrole and condensation with 4-methoxy-2,2'-bipyrrole-5-carboxyaldehyde to form prodigiosin, as we have shown for Serratia sp. ATCC 39006.


A tale of two tails: cytosolic termini and K(+) channel function.
The enormous variety of neuronal action potential waveforms can be ascribed, in large part, to the sculpting of their falling phases by currents through voltage-gated potassium channels. These proteins play several additional roles in other tissues such as the regulation of heartbeat and of insulin release from pancreatic cells as well as auditory signal processing in the cochlea. The functional channel is a tetramer with either six or two transmembrane segments per monomer. Selectivity filters, voltage sensors and gating elements have been mapped to residues within the transmembrane region. Cytoplasmic residues, which are accessible targets for signal transduction cascades and provide attractive means of regulation of channel activity, are now seen to be capable of modulating various aspects of channel function. Here we review structural studies on segments of the cytoplasmic tails of K(+) channels, as well as the range of modulatory activities of these tails.


Genotype-environment interactions in determining fitness in dense, artificial populations of Phlox drummondii.
Survivorship and growth of individuals of the annual plant Phlox drummondii were examined in artificial, density-stressed populations composed of five genetically differentiated varieties sown in monocultures and in five pairwise mixtures. All experiments were replicated under two treatments of nutrient availability.Varieties differed significantly in competitive ability, and relative competitive success was shown to be highly habitat (treatment) dependent. In 19 of 20 mixed populations, the relative rankings of pairs of competitors were consistent with a priori expectations based on the morphological and/or historical differences between seed sources, but were not predictable from relative tolerances to density stress in monoculture.Estimated selection coefficients based on relative competitive abilities ranged from 0.04 to 0.94. Despite these large fitness differences among competing varieties, most of the variance in the absolute fitness of individuals resided within varieties. This variation may be mostly environmental.In the high nutrient treatment, where competition for light was most intense, variation in individual plant size was lower in mixed culture as compared to monocultures. Genetic variation for competitive ability may actually lead to greater size uniformity among survivors by promoting an early initiation of thinning, thereby reducing the density stress on survivors.


Hemorrhagic disease in dogs infected with an unclassified intraendothelial piroplasm in southern Brazil.
A hemorrhagic disease affecting dogs in Brazil, referred to popularly as "nambiuvú" (bloody ears) and believed to be transmitted by ticks, has been observed in animals infected with an organism described originally in 1910 as a piroplasm, and known locally as Rangelia vitalii. In this series of 10 cases, the disease was characterized by anaemia, jaundice, fever, spleno- and lymphadenomegaly, hemorrhage in the gastrointestinal tract, and persistent bleeding from the nose, oral cavity and tips, margins and outer surface of the pinnae. The ixodid ticks Rhipicephalus sanguineus and Amblyomma aureolatum infested affected dogs from suburban and rural areas, respectively. Laboratory findings included regenerative anaemia, spherocytosis, icteric plasma and bilirubinuria. Those intracellular organisms were found in bone marrow smears but not in blood smears. Microscopically, zoites were seen within the cytoplasm of blood capillary endothelial cells. Parasitized and non-parasitized endothelial cells were positive immunohistochemically for von Willebrand factor (vWF). Langhans-type multinucleate giant cells were observed in the lymph nodes and choroid plexus. There was prominent erythrophagocytosis by macrophages in the lymph node sinuses and infiltration of the medullary cords by numerous plasma cells. Ultrastructurally, this organism had an apical complex that included a polar ring and rhoptries but no conoid. This parasite was contained within a parasitophorous vacuole that had a trilaminar membrane with villar protrusions and was situated in the cytoplasm of capillary endothelial cells. This organism tested positive by immunohistochemistry for Babesia microti. This pathogen was also positive by in situ hybridization for B. microti. Tentative clinical diagnosis in these cases was based on the history, clinical picture, haemogram and favorable response to therapy, and confirmed through microscopic examination of smears from the bone marrow or histological sections of multiple tissues, especially lymph nodes where zoites were most frequently found. The disease was reproduced by intravenous inoculation of blood from a naturally infected dog into an experimental dog. The authors demonstrate in this study that this organism is a protozoa of the phylum Apicomplexa, order Piroplasmorida. This piroplasm seems to be different from Babesia since it has an intraendothelial stage. Molecular phylogenetic analysis is necessary to better characterize this parasite and clarify its taxonomic status.


Pharmacokinetics of 5-fluorouracil: inter-relationship with biochemical kinetics in monitoring therapy.
The major difficulty in using pharmacokinetic concepts for monitoring individual therapy with 5-fluorouracil is based on the mostly intracellular location of the active nucleotide metabolites of 5-fluorouracil with no clear correlation of plasma levels of the drug. In addition, the basic biochemical mechanism of 'thymine-less cell death' following inhibition of de novo thymidylate synthesis by 2'-deoxy-5-flurouridine 5'-monophosphate (FdUMP) is poorly understood, and only some of the biochemical determinants of therapeutic response to 5-fluorouracil are known. Individualised therapy with 5-fluorouracil requires an intergrated approach which should include methods of pharmacokinetics and biochemical kinetics. 5-Fluorouracil stands as an example for most of the pyrimidine and purine metabolites to which similar consideration apply in monitoring of cancer chemotherapy.


Evaluation of honey-baited FTA cards in combination with different mosquito traps in an area of low arbovirus prevalence.
The threat of mosquito-borne diseases is increasing in continental Europe as demonstrated by several autochthonous chikungunya, dengue and West Nile virus outbreaks. In Switzerland, despite the presence of competent vectors, routine surveillance of arboviruses in mosquitoes is not being carried out, mainly due to the high costs associated with the need of a constant cold chain and laborious processing of thousands of mosquitoes. An alternative approach is using honey-baited nucleic acid preserving cards (FTA cards) to collect mosquito saliva that may be analysed for arboviruses. Here, we evaluate whether FTA cards could be used to detect potentially emerging viruses in an area of low virus prevalence in combination with an effective mosquito trap. In a field trial in southern Switzerland we measured side-by-side the efficacy of the BG-Sentinel 2, the BG-GAT and the Box gravid trap to catch Aedes and Culex mosquitoes in combination with honey-baited FTA cards during 80 trapping sessions of 48 hours. We then screened both the mosquitoes and the FTA cards for the presence of arboviruses using reverse-transcription PCR. The efficacy of the compared trap types was evaluated using generalized linear mixed models. The Box gravid trap collected over 11 times more mosquitoes than the BG-GAT and BG-Sentinel 2 trap. On average 75.9% of the specimens fed on the honey-bait with no significant difference in feeding rates between the three trap types. From the total of 1401 collected mosquitoes, we screened 507 Aedes and 500 Culex females for the presence of arboviruses. A pool of six Cx. pipiens/Cx. torrentium mosquitoes and also the FTA card from the same Box gravid trap were positive for Usutu virus. Remarkably, only two of the six Culex mosquitoes fed on the honey-bait, emphasising the high sensitivity of the method. In addition, two Ae. albopictus collections but no FTA cards were positive for mosquito-only flaviviruses. Based on our results we conclude that honey-baited FTA cards, in combination with the Box gravid trap, are an effective method for arbovirus surveillance in areas of low prevalence, particularly where resources are limited for preservation and screening of individual mosquitoes.


Investigation of response factors for monotherapy with immune checkpoint inhibitors in non-small cell lung cancer patients with PD-L1 expression <50.
Immune checkpoint inhibitor (ICI) monotherapy is currently approved for the treatment of advanced non-small cell lung cancer (NSCLC) patients with programmed death ligand-1 (PD-L1) expression ≥50%. However, the efficacy of ICI monotherapy in patients with PD-L1 expression <50% has not yet been fully elucidated. The aim of this study was to identify the clinical characteristics of NSCLC patients with PD-L1 expression <50% who respond to single-agent ICIs and factors that predict response. Patients with advanced or recurrent NSCLC with a PD-L1 tumor proportion score (TPS) of 50% or less who received new monotherapy with an ICI between July 2012 and December 2022 were retrospectively analyzed. Patients with response were compared with those without response in the post-treatment response assessment. Among the 37 patients, six (16.2%) NSCLC patients in the response group responded to ICI monotherapy and had a significantly lower body mass index (BMI) (p = 0.003). Significantly more patients in the response group developed immune-related adverse events (irAEs) than in the nonresponse group (p < 0.001). Multivariate analysis identified high BMI as a significant independent risk factor predicting nonresponse to ICI monotherapy in NSCLC patients with PD-L1 < 50%. Among NSCLC patients with PD-L1 < 50%, those with a higher BMI were more likely to be nonresponders to ICI monotherapy. In addition, the group that responded to ICI monotherapy may have been at higher risk of developing irAEs, suggesting that careful follow-up is warranted.


The role of sleep electroencephalography in patients with new onset epilepsy.
An increased propensity for seizures is associated with different stages of the sleep-wake cycle. In this study, we prospectively analyzed patients with new-onset epilepsy and investigated the clinical correlates of the yield obtained from sleep electroencephalography (EEG) recordings in patients with a normal wakefulness EEG. All patients admitted to our epilepsy unit due to unprovoked epileptic seizures and not yet treated with antiepileptic drugs were recruited consecutively for the last three years. All had a routine EEG at wakefulness (WEEG), and those with no epileptiform activity had a video-EEG recording during sleep (SEEG). We investigated a total of 241 patients; 129 patients (53.5%) had both wakefulness and sleep EEG recordings. The patients with abnormal WEEG were older than those with normal WEEG (p = 0.005). Abnormal WEEG was detected in only 31.2% of patients with focal seizures, but in 77.3% of patients with generalized seizures (p < 0.001). WEEG was abnormal in 44.0% of patients with diurnal seizures, but in 27.5% of nocturnal seizures (p = 0.007). Abnormal WEEG was present in 75.5% of patients with a presumed genetic origin and in 59.3% of patients with structural etiology (p < 0.001). Sleep EEG detected an abnormality in 41.8% of patients with normal WEEG; of these, 82.8% were focal abnormalities. In contrast, the majority of abnormalities detected in WEEG were generalized (55.8%, p < 0.001). Our results showed a greater likelihood of abnormal WEEG in older patients and in those with generalized epilepsy, diurnally precipitating seizures, and epilepsy of presumed genetic origin.


Implementation of lung cancer screening in Europe: challenges and potential solutions: summary of a multidisciplinary roundtable discussion.
Recent randomised trials on screening with low-dose CT have shown important reductions in lung cancer (LC) mortality and have triggered international efforts to implement LC screening. Detection rates of stage I LC with volume CT approaching 70% have been demonstrated. In April 2019 'ESMO Open - Cancer Horizons' convened a roundtable discussion on the challenges and potential solutions regarding the implementation of LC screening in Europe. The expert panel reviewed the current evidence for LC screening with low-dose CT and discussed the next steps, which are covered in this article. The panel concluded that national health policy groups in Europe should start to implement CT screening as adequate evidence is available. It was recognised that there are opportunities to improve the screening process through 'Implementation Research Programmes'.


Gorham's syndrome: a case report and review of the literature.
Gorham's syndrome is a rare disorder involving a proliferation of vascular channels associated with extensive loss of bony matrix. A case report is presented with a review of the 97 previously reported cases. The age of patients at presentation has ranged from less than one to 75 years (mean: 27 years). Sixty-four percent have been men. Fifty-seven percent have had a history of prior trauma. Laboratory values for systemic measures have usually been normal. The disease usually arrests spontaneously, but this is unpredictable. Sixteen patients (16 percent) have died of the disorder, with 10 deaths due to chest wall involvement, three to spinal cord transection, two to sepsis, and one to asphyxia and aspiration. Although the mechanism of bone loss is unknown, osteoclasts were focally increased in the case described herein. Further information and investigation are needed to better understand this unusual disorder.


Escherichia coli recombinant sperm immobilizing factor RecX as a potential vaginal contraceptive.
To control the overpopulation and unintended pregnancies, vaginal contraceptives have gained recent surge of interest because of its topical application with possible avoidance of systemic effects. However non-specific cytotoxicity associated with detergent-based synthetic vaginal contraceptive agents limits their use and generates considerable interest in the development of vaginal contraceptives of biological origin for controlling reproduction and ultimately growing population. In this study, we have cloned, over-expressed an Escherichia coli gene encoding a sperm immobilizing factor (SIF) that inhibits sperm motility for the development of vaginal contraceptive from a biological source i.e. E. coli. The contraceptive efficacy of the Escherichia coli recombinant sperm immobilizing factor (r-SIF) was also determined. Genomic DNA library of an E. coli strain isolated from semen sample of an infertile male was constructed for the identification and cloning of E. coli SIF coding gene. This gene was sub-cloned in pBADmycHisB for over-expression and the r-SIF was purified using Ni-NTA affinity chromatography. Effect of r-SIF on mouse sperm motility, viability and on morphology was evaluated. Binding of r-SIF to mouse sperm was demonstrated by fluorescent labeling. Contraceptive efficacy of r-SIF was checked in murine model. Genomic library resulted in five hundred transformants; five clones were found positive for sperm immobilizing activity. The protein product of the insert DNA sequence in one of the transformants showed maximum sperm immobilizing activity. Sequence analysis of ORFs in the insert revealed homology to recX on both nucleotide and protein level. 40 μg of the purified r-SIF showed immediate spermicidal activity in vitro for mouse sperm. Scanning electron micrograph of the r-SIF treated sperm showed intense morphological damage to sperm. FITC labeled r-SIF showed highest fluorescence at the head region of the sperm. 5 μg of purified r-SIF exhibited a complete contraceptive effect in mouse model. r-SIF could be seen as potential target to be developed as potent and safe vaginal contraceptive in future.


Promoter-proximal CCCTC-factor binding is associated with an increase in the transcriptional pausing index.
It has been known for more than 2 decades that after RNA polymerase II (RNAPII) initiates transcription, it can enter into a paused or stalled state immediately downstream of the transcription start site before productive elongation. Recent advances in high-throughput genomic technologies facilitated the discovery that RNAPII pausing at promoters is a widespread physiologically regulated phenomenon. The molecular underpinnings of pausing are incompletely understood. The CCCTC-factor (CTCF) is a ubiquitous nuclear factor that has diverse regulatory functions, including a recently discovered role in promoting RNAPII pausing at splice sites. In this study, we analyzed CTCF binding sites and nascent transcriptomic data from three different cell types, and found that promoter-proximal CTCF binding is significantly associated with RNAPII pausing.


Orbital-angular-momentum crosstalk and temporal fading in a terrestrial laser link using single-mode fiber coupling.
Using a mobile experimental testbed, we perform a series of measurements on the detection of laser beams carrying orbital angular momentum (OAM) to evaluate turbulent channel distortions and crosstalk among receive states in an 84-m roofed optical link. We find that a receiver assembly using single-mode fiber coupling serves as a good signal selector in terms of crosstalk rejection. From the recorded temporal channel waveforms, we estimate average crosstalk profiles and propose an appropriate probability density function for the fluctuations of the detected OAM signal. Further measurements of OAM crosstalk are described for a horizontal 400-m link established over our campus.


Criteria for Applying the Lucas-Washburn Law.
Spontaneous imbibition happens in many natural and chemical engineering processes in which the mean advancing front usually follows Lucas-Washburn's law. However it has been found that the scaling law does not apply in many cases. There have been few criteria to determine under what conditions the Washburn law works. The effect of gravity on spontaneous imbibition in porous media was investigated both theoretically and experimentally. The mathematical model derived analytically was used to calculate the imbibition rates in porous media with different permeabilities. The results demonstrated that the effect of gravity on spontaneous imbibition was governed by the hydraulic conductivity of the porous media (permeability of the imbibition systems). The criteria for applying the Lucas-Washburn law have been proposed. The effect of gravity becomes more apparent with the increase in permeability or with the decrease in CGR number (the ratio of capillary pressure to gravity forces) and may be ignored when the CGR number is less than a specific value N(*)(cg) ≅ 3.0. The effect of gravity on imbibition in porous media can be modeled theoretically. It may not be necessary to conduct spontaneous imbibition experiments horizontally in order to exclude the effect of gravity, as has been done previously.


Analysis of heterochromatin by combination of C-banding and CMA3 and DAPI staining in two fish species (Pimelodidae, Siluriformes).
The chromosomes of Steindachneridion sp. (2n = 56) and Rhamdia quelen (2n = 58) were analyzed by C-banding (CB) and Chromomycin A3 (CMA3) and 4,6-diamidino-2-phenylindole (DAPI) staining, separately and consecutively, in order to understand the role of base-specific fluorochrome treatment after CB. Both species' chromosomes shared common staining profiles as follows. CB with Giemsa (CBG) revealed weak heterochromatic blocks in the telomeric regions of some chromosomes and conspicuous bands on the short arms of one chromosome pair, where nucleolar organizer regions (NORs) were evidenced by silver-staining. Without CB pretreatment, the NORs were stained conspicuously with CMA3, but not with DAPI. The latter uniformly stained all chromosomes, but leaving the NORs pale. Combination of CMA3 or DAPI staining with CB showed distinctive fluorescent blocks in the NOR-bearing short arms of the single chromosome pair along with several bright fluorescent signals on other chromosomes, which were not evidenced by single CMA3 or DAPI staining. These results suggest a modification of chromatin structure by CB treatment, which may increase the stainability of CMA3 and DAPI.


Benign vulvar tumors.
Benign tumors and "dark lesions" accounted for 22% of vulvar disease seen in the Vulvar Clinic at the Milwaukee County Medical Complex over an 8-year period. Biopsy confirmation was obtained for 269 lesions. The order of frequency of lesions in this study was as follows: epidermal inclusion cyst, lentigo, Bartholin's duct obstruction, carcinoma in situ, melanocytic nevi, acrochordon, mucous cyst, hemangiomas, postinflammatory hyperpigmentation, seborrheic keratoses, varicosities, hidradenomas, verruca, basal cell carcinoma, and, last, unusual tumors such as neurofibromas, ectopic tissue, syringomas, and abscesses. The variability in clinical appearance of vulvar tumors suggests that biopsy confirmation should be obtained on all lesions for which there is the least doubt regarding the diagnosis.


Internal Transcribed Spacer Sequence-Based Phylogeny and Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Differentiation of Tilletia walkeri and T. indica.
ABSTRACT A polymerase chain reaction-restriction fragment length polymorphism assay to distinguish Tilleita walkeri, a rye grass bunt fungus that occurs in the southeastern United States and Oregon, from T. indica, the Karnal bunt fungus, is described. The internal transcribed spacer (ITS) region of the ribosomal DNA repeat unit was amplified and sequenced for isolates of T. indica, T. walkeri, T. horrida, and a number of other taxa in the genus Tilletia. A unique restriction digest site in the ITS1 region of T. walkeri was identified that distinguishes it from the other taxa in the genus. Phylogenetic analysis of the taxa based on ITS sequence data revealed a close relationship between T. indica and T. walkeri, but more distant relationships between these two species and other morphologically similar taxa.


FeS combined ozonation to remove p-aminobenzenesulfonamide from water: Density functional theory insights into the mechanism.
The applicability and performance of FeS in ozonation process to remove p-aminobenzenesulfonamide (SN) from water was assessed, and the working mechanism of FeS was comprehensively explored by both experimental means and density functional theory (DFT) simulation. FeS combined ozonation achieved 74% of SN removal in 60 min under the optimal condition, which was 37% higher than by ozonation alone, and 12% higher than FeO combined ozonation. Highly active species of •OH, •SO4-, 1O2 and •O2- were detected in the FeS combined ozonation system, the evolution pathway of the involved species was expounded with the aid of DFT calculation. The results revealed that •O2-, H2O2 and SO42- were originally formed via interface reactions on FeS surface, then gradually transformed into •OH, 1O2 and •SO4- through subsequent chain reactions. Moreover, FeS had a lower energy barrier of 0.16 eV than FeO with a value of 0.83 eV for the transformation of ozone to active atomic oxygen. The presented study provided a significant insight into the role of Fe-based materials in ozonation, and was of great importance to guide the route for ozonation process improvement.


Ground-state decay rate for the Zener breakdown in band and Mott insulators.
Nonlinear transport of electrons in strong electric fields, as typified by dielectric breakdown, is reformulated in terms of the ground-state decay rate originally studied by Schwinger in nonlinear QED. We discuss the effect of electron interaction on Zener tunneling by comparing the dielectric breakdown of the band insulator and the Mott insulator, where the latter is studied by the time-dependent density-matrix renormalization group. The relation with the Berry's phase theory of polarization is also established.


Natural photosystems from an engineer's perspective: length, time, and energy scales of charge and energy transfer.
The vast structural and functional information database of photosynthetic enzymes includes, in addition to detailed kinetic records from decades of research on physical processes and chemical reaction-pathways, a variety of high and medium resolution crystal structures of key photosynthetic enzymes. Here, it is examined from an engineer's point of view with the long-term goal of reproducing the key features of natural photosystems in novel biological and non-biological solar-energy conversion systems. This survey reveals that the basic physics of the transfer processes, namely, the time constraints imposed by the rates of incoming photon flux and the various decay processes allow for a large degree of tolerance in the engineering parameters. Furthermore, the requirements to guarantee energy and electron transfer rates that yield high efficiency in natural photosystems are largely met by control of distance between chromophores and redox cofactors. This underlines a critical challenge for projected de novo designed constructions, that is, the control of spatial organization of cofactor molecules within dense array of different cofactors, some well within 1 nm from each other.


Similarities and differences between behavioral control by drug-produced stimuli and by sensory stimuli.
Drug-induced state-dependent learning and drug discriminations may be based on sensory stimuli induced by drug actions, or at least on events in the brain that have properties analogous to those of sensory stimuli. A variety of comparisons between behavioral control by drug-induced stimuli and by classically defined interoceptive and exteroceptive stimuli are possible. These allow inferences to be made about the nature of drug stimuli, the properties of behavioral paradigms that we use to investigate them, and the mechanisms by which drugs achieve behavioral control in drug discrimination and state-dependent learning paradigms. Each of these topics is selectively reviewed in this paper.


The infant with nystagmus, normal appearing fundi, but an abnormal ERG.
Many retinal disorders present during infancy with nystagmus, decreased vision, and normal-appearing fundi, but an abnormal ERG. The most common of these disorders are Leber's congenital amaurosis, achromatopsia, and congenital stationary night-blindness. Other disorders with similar ocular manifestations may be associated with a variety of life-threatening systemic abnormalities. This review describes the clinical, electrophysiological, and laboratory findings that can be used to distinguish among these conditions.


An extended approach for the generalized powered uniform distribution.
A new uniform distribution model, generalized powered uniform distribution (GPUD), which is based on incorporating the parameter k into the probability density function (pdf) associated with the power of random variable values and includes a powered mean operator, is introduced in this paper. From this new model, the shape properties of the pdf as well as the higher-order moments, the moment generating function, the model that simulates the GPUD and other important statistics can be derived. This approach allows the generalization of the distribution presented by Jayakumar and Sankaran (2016) through the new GPUD ( J - S ) distribution. Two sets of real data related to COVID-19 and bladder cancer were tested to demonstrate the proposed model's potential. The maximum likelihood method was used to calculate the parameter estimators by applying the maxLik package in R. The results showed that this new model is more flexible and useful than other comparable models.


Ecosystem service deficits of European cities.
Climate change and biodiversity loss are two pressing global environmental challenges that are tightly coupled to urban processes. Cities emit greenhouse gases through the consumption of materials and energy. Urban expansion encroaches on local habitats, while urban land teleconnections simultaneously degrade distant ecosystems. These processes decrease the supply of and increase the demand for ecosystem services inside and outside urban areas. Most cities are in a state of ecosystem services deficit, whereby demand exceeds local supply of ecosystem services. Methods to quantify this deficit by capturing multi-scale and multi-level ecological exchanges are incipient, leaving scholars with a partial understanding of the environmental impacts of cities. This paper deploys a novel method to simulate future urban supplies and demands of two key ecosystem services needed to combat climate change and biodiversity loss - global climate regulation and global habitat maintenance. Applying our model to eight representative European cities, we project growing ecosystems deficits (demand exceeds supply) between 8% and 214% in global climate regulation and 11% and 431% in global habitat maintenance between 2020 and 2050. Variation between cities stems from differing dietary patterns and electricity mixes, which have large implications for ecosystems outside the city. To combat these losses, urban sustainability strategies should complement local restoration with changes to local consumption alongside promoting remote ecological restoration to tackle the multi-level environmental impacts of cities.


Exocytosis and Spreading of Normal and Aberrant α-Synuclein.
It is now established that α-synuclein can be physiologically secreted to the extracellular space. In this sense, mechanisms that govern the secretion of the protein may be of importance in the initiation and progress of synucleinopathies. It is possible that increased secretion may aid the formation of toxic seeds extracellularly. Alternatively, reduced presence of extracellular α-synuclein due to impaired secretion may increase the intracellular load and trigger intracellular seeding. Once outside, α-synuclein can exert various paracrine actions on neighboring cells again by mechanisms that have not been fully elucidated. It has been demonstrated that, when applied extracellularly, α-synuclein species can induce multiple neurotoxic and inflammatory responses, and aid the transmission of pathology between neurons. Still, the exact mechanism(s) by which secreted α-synuclein affects the homeostasis of other neurons is still not well understood. A portion of α-synuclein has been shown to be associated with the surface and lumen of exosomes which can transfer it to the surrounding cells, and potentially trigger seeding. Interestingly, increased exosome release has been linked to pathological situations of lysosomal dysfunction as observed in Parkinson's disease (PD). However, the possibility that the observed α-synuclein pathology spread is attributable to the passive diffusion of the initial injected α-synuclein strains cannot be excluded. Importantly, most of the studies that have so far addressed the role of extracellular α-synuclein have not employed naturally secreted forms of the protein. It is plausible that deregulation in the normal processing of secreted α-synuclein may aid the formation of "toxic" species and as such it may also be a causative risk factor for PD. In this capacity, elucidation of the underlying mechanisms that regulate the protein-levels of extracellular α-synuclein becomes essential. Such mechanisms could involve its proteolytic clearance from the extracellular milieu.


Electronic imaging system for direct and rapid quantitation of fluorescence from electrophoretic gels: application to ethidium bromide-stained DNA.
We have built an electronic imaging system based on a modified charge-coupled-device television camera that directly quantitates the distribution of fluorescence from electrophoretic gels, chromatograms, and other stationary sources. Exposure times can exceed 1 min. Unlike the photographic system that it replaces, the response of the camera is directly proportional to the intensity of incident fluorescence, and image data are digitized and stored in computer memory ready for analysis immediately upon completion of an exposure. We describe procedures for the display, normalization, and archival storage of image data and programs that use images of ethidium bromide-stained DNA in alkaline agarose gels to quantitate single-strand breaks in DNA.


Metastability and polymorphism in the gel phase of 1,2-dipalmitoyl-3-SN-phosphatidylcholine. A Fourier transform infrared study of the subtransition.
Fourier transform infrared spectroscopy was used to study the metastability of 1,2-dipalmitoyl-3-sn-phosphatidylcholine (DPPC) at temperatures near 0 degrees C. It was found that when DPPC is incubated at 2 degrees C for three days the two-dimensional acyl chain packing changes from one resulting in spectra typical of an orthorhombic subcell to one resembling that found in triclinically packed acyl systems. This transition proceeds in two stages. The first step, requiring less than one day, approximates first-order kinetics; the second stage proceeds with second- or higher-order kinetics. Comparison of spectra recorded at -36 degrees C with and without prior incubation at 2 degrees C shows that there are two stable low temperature forms of DPPC; that is, DPPC is metastable only within a narrow temperature range. A study of the thermotropic behavior in the range 0-45 degrees C shows that the subtransition near 15 degrees C is a transition from the alternate form to one with orthorhombic characteristics. Spectral changes at the pretransition and the main phase transition demonstrate that there are differences in behavior that are related to the thermal history of the sample.


α-1,6-Fucosyltransferase (FUT8) inhibits hemoglobin production during differentiation of murine and K562 human erythroleukemia cells.
Erythropoiesis results from a complex combination of the expression of several transcription factor genes and cytokine signaling. However, the overall view of erythroid differentiation remains unclear. First, we screened for erythroid differentiation-related genes by comparing the expression profiles of high differentiation-inducible and low differentiation-inducible murine erythroleukemia cells. We identified that overexpression of α-1,6-fucosyltransferase (Fut8) inhibits hemoglobin production. FUT8 catalyzes the transfer of a fucose residue to N-linked oligosaccharides on glycoproteins via an α-1,6 linkage, leading to core fucosylation in mammals. Expression of Fut8 was down-regulated during chemically induced differentiation of murine erythroleukemia cells. Additionally, expression of Fut8 was positively regulated by c-Myc and c-Myb, which are known as suppressors of erythroid differentiation. Second, we found that FUT8 is the only fucosyltransferase family member that inhibits hemoglobin production. Functional analysis of FUT8 revealed that the donor substrate-binding domain and a flexible loop play essential roles in inhibition of hemoglobin production. This result clearly demonstrates that core fucosylation inhibits hemoglobin production. Third, FUT8 also inhibited hemoglobin production of human erythroleukemia K562 cells. Finally, a short hairpin RNA study showed that FUT8 down-regulation induced hemoglobin production and increase of transferrin receptor/glycophorin A-positive cells in human erythroleukemia K562 cells. Our findings define FUT8 as a novel factor for hemoglobin production and demonstrate that core fucosylation plays an important role in erythroid differentiation.


Faster improvement in outcome scores in posterior stabilised total knee arthroplasty compared to medial congruent system with posterior cruciate ligament retained.
Most studies comparing medial pivot to the posterior stabilised (PS) systems sacrifice the PCL. It is unknown whether retaining the PCL in the Medial Congruent (MC) system may provide further benefit compared to the more commonly used PS system. A retrospective review of a single-surgeon's registry data comparing 44 PS and 26 MC with PCL retained (MC-PCLR) TKAs was performed. Both groups had similar baseline demographics. The PS and MC-PCLR groups had similar pre-operative range of motion (ROM) (PS:104º ± 20º vs. MC-PCLR: 101º ± 19º, p = 0.70), Oxford Knee Score (OKS) (PS: 27 ± 6 vs. MC-PCLR: 26 ± 7, p = 0.62), and Knee Society Scoring System (KS) Function Score (KS-FS) (PS: 52 ± 24 vs. MC-PCLR: 56 ± 23, p = 0.49). The pre-operative KS Knee Score (KS-KS) was significantly lower in the PS group (PS: 44 ± 14 vs. MC-PLR: 53 ± 18, p < 0.05). At 12-months post-operation, there was significant improvement in all parameters (p < 0.01). Both groups had similar ROM (PS: 115º ± 13º vs. MC-PCLR: 114º ± 10º, p = 0.98), OKS (PS: 41 ± 5 vs. MC-PCLR: 40 ± 4, p = 0.50), KS-FS (PS: 74 ± 22 vs. MC-PCLR: 77 ± 16, p = 0.78), and KS-KS (PS: 89 ± 10 vs. MC-PCLR: 89 ± 10, p = 0.89). The PS group had significant improvement in all parameters from preoperation to 3-month postoperation (p < 0.05), but not from 3-month to 1-year postoperation (p ≥ 0.05). The MC-PCLR group continued to have significant improvement from 3-month to 1-year postoperation (p < 0.05). Preserving the PCL when using MC may paradoxically cause an undesired additional restrain that slows the recovery process of the patients after TKA. Compared to MC-PCLR, a PS TKA may expect significantly faster improvement at 3 months post operation, although they will achieve similar outcomes at 1-year post operation.


Big Data/AI in Neurocritical Care: Maybe/Summary.
Big data (BD) and artificial intelligence (AI) have increasingly been used in neurocritical care. "BD" can be operationally defined as extremely large datasets that are so large and complex that they cannot be analyzed by using traditional statistical modeling. "AI" means the ability of machines to perform tasks similar to those performed by human intelligence. We present a brief overview of the most commonly applied AI techniques to perform BD analytics and discuss some of the recent promising examples in the field of neurocritical care. The latter include the following: cognitive motor dissociation in disorders of consciousness, hypoxic-ischemic injury following cardiac arrest, delayed cerebral ischemia and vasospasm after subarachnoid hemorrhage, and monitoring of intracranial pressure. It is imperative that we develop multicenter collaborations to tackle BD. These collaborations will allow us to share data, combine predictive algorithms, and analyze multiple and cumulative sources of data retrospectively and prospectively. Once AI algorithms are validated at multiple centers, they should be tested in randomized controlled trials investigating their impact on clinical outcome. The neurocritical care community must work to ensure that AI incorporates standards to ensure fairness and health equity rather than reflect our biases present in our collective conscience.


In vitro comparison of antimicrobial effectiveness of QMix and other final irrigants in human root canals.
Final root canal irrigation stands as an effective strategy for eliminating the dentin infection. This study aimed to investigate and compare the antibacterial efficacy of QMix and other four final irrigation regimens in reducing Enterococcus faecalis within human root canals. Single-canal human teeth contaminated with E. faecalis for 4 weeks were prepared chemomechanically with sodium hypochlorite (NaOCl). Then, the teeth were randomly assigned into six groups according to the final irrigation protocols: (1) EDTA/NaOCl, 17% EDTA followed by 5.25% NaOCl; (2) EDTA/chlorhexidine (CHX), 17% EDTA followed by 2% CHX; (3) EDTA/cetrimide (CTR), 17% EDTA followed by 2% CTR; (4) MTAD; (5) QMix; and (6) control, 0.9% saline. Bacterial samples collected before instrumentation and after final irrigation were cultured and the colony-forming units (CFUs) were counted. The CFUs in the QMix, EDTA/CHX, and EDTA/CTR groups were significantly lower than those in the EDTA/NaOCl group. No significant differences were observed between the QMix, EDTA/CHX, and EDTA/CTR groups. MTAD showed weaker ability than QMix and EDTA/CHX to eliminate E. faecalis, but it caused a greater reduction in CFU than EDTA/NaOCl. Hence, the antimicrobial activity of QMix was comparable to that of EDTA/CHX and EDTA/CTR and more effective than that of EDTA/NaOCl against intracanal E. faecalis.


Extraction of arsenate and arsenite species from soils and sediments.
The primary objective of this study was to develop a simple method that can be used to extract the more readily mobilizable and bioavailable arsenic species from soil and sediment while at the same time minimizing the transformation between (AsIII) and (AsV), the two most commonly found arsenic species in the environment. Several extraction strategies were evaluated using phosphate as extractant in combination with either ethylenediaminetetraacetic acid (EDTA), hydroxylamine hydrochloride (NH2OH.HCl), or sodium diethyldithiocarbamate trihydrate (NaDDC). The addition of EDTA in the phosphate solution did not prevent AsIII from oxidation. While promising results were shown when 1% NH2OH.HCl was added, conversion of AsIII began to occur with extended extraction time (> 12 h). Good results were achieved using 10 mM phosphate and 0.5% NaDDC where AsIII oxidation was clearly minimized. The combined phosphate and NaDDC solution was applied to several soil and sediment samples. AsIII spiked was quantitatively recovered in all soil types tested.


The JAK2V617F tyrosine kinase mutation has no impact on overall survival and the risk of leukemic transformation in myelofibrosis.
The prevalence of JAK2V617F tyrosine kinase mutation differs between various variants of myelofibrosis with the higher detection rate for patients with post-polycythemia vera myelofibrosis (post-PV MF; 91%) if compared to primary myelofibrosis (PMF; 45%) and post-essential thrombocythemia myelofibrosis (post-ET MF; 39%). The impact of V617F point mutation and its allele burden on overall survival (OS) and the risk of leukemic transformation (LT) has been the subject of several studies, but the results were ambiguous. Our study included 77 patients with the following variants: 42 patients with PMF (55%), 16 with post-ET MF (21%) and 19 with post-PV MF (24%). Median age at diagnosis for the entire cohort was 61 years (range 19-81), with 53% of female. A total of 42 patients were JAK2V617F positive, giving an overall frequency of 55%; the median allele burden was 22% (range 2-96%). The JAK2V617F point mutation was detected in 21 patients with PMF (50%), 14 with post-PV MF (88%) and 7 with post-ET MF (37%). Lower JAK2V617F allele burden was more frequently detected in PMF patients, whereas higher allele burden was predominantly seen in post-PV/ET MF group. There was no significant difference between V617F-positive and V617F-negative patients in terms of studied parameters in PMF as well as in post-PV/ET MF subgroup. No significant difference was also demonstrated when the above-mentioned subpopulations were analyzed according to JAK2V617F allele burden, except higher leukocyte count in post-PV/ET MF patients with higher allele burden (14.3×10(9)/L vs. 6.2×10(9)/L; p=.03). Median follow-ups for V617F-positive and V617F-negative patients were 16.6 months (range 3.6-206.4) and 36.4 months (range 2.5-142.1), respectively. The presence of JAK2V617F mutation did not affect OS and the risk of LT development.


Prevalence of arsenic-induced skin lesions and associated factors in Ethiopia: Community-based study.
The study aimed to assess the prevalence of arsenic-induced skin lesions and associated factors among the population in the Adami Tulu Jido Kombolcha district, Ethiopia. A community-based cross-sectional study design was employed among 403 participants from June 02-20, 2022. A two-stage cluster sampling method was conducted to select study subjects. A pretested structured and semi-structured interview questionnaire and observation using a WHO flow chart were used for data collection. Data analysis was performed using SPSS version 24 statistical software for Windows. A multivariable binary logistic regression model was applied to examine the relationship between predictor variables and an outcome variable. The degrees of association between outcomes and predictor variables were assessed using ORs and 95% CIs, and P-values < 0.05 were considered significant. The prevalence of arsenic-induced skin lesions (arsenicosis) in the study area was 2.2% [95% CI: 1.0-3.7]. The most common arsenic-induced manifestation was keratosis (55.6%), followed by hyperpigmentation (33.3%) and hyperkeratosis (11.1%). Consumption of well water, smoking cigarettes, and chewing khat were significantly associated with arsenic-induced skin lesions. Therefore, the findings of this study should trigger further research on arsenic exposure and health risks.


Spatial statistical modeling of disease outbreaks with particular reference to the UK foot and mouth disease (FMD) epidemic of 2001.
In this paper we examine issues relating to the analysis of spatially-referenced disease data. Initially, we discuss the use of exploratory statistical tools such as density estimation and nonparametric regression. We then consider the need for descriptive epidemic models in space, time, and space-time models for epidemic dynamics. Implicitly space-time must be considered in any analysis of the spatial structure of epidemics. The use of Bayesian models for disease spread is discussed and applied to the recent foot and mouth outbreak in the UK.


Opposite beta2-glycoprotein I requirement for the binding of infectious and autoimmune antiphospholipid antibodies to cardiolipin liposomes is associated with antibody avidity.
The aim of this study was to investigate the interaction of antiphospholipid antibodies (aPL) from two different populations (patients with autoimmune or infectious disorders) with cardiolipin (CL) arranged in a defined bilayer. beta2-Glycoprotein I (beta2GPI), an apolipoprotein that plays a critical role in the aPL binding to phospholipids, was quantified by dot blot in purified IgG-aPL samples, further classified according to apparent avidity to CL. In solid-phase assays, beta2GPI increased, preferentially, the binding of low-avidity autoimmune aPL to CL but inhibited the binding of low-avidity syphilitic aPL. In the absence of beta2GPI, both autoimmune and infectious aPL induced the leakage of the entrapped fluorescent probe, carboxyfluorescein (CF), from small unilamellar vesicles containing CL. aPL-induced probe leakage was protein concentration-dependent and characterized by a lag-phase onset of 100-120 min. beta2GPI increased the leakage rate induced by low-avidity autoimmune aPL only and inhibited the leakage induced by all syphilitic aPL. The following conclusions were provided: (1) in the absence of beta2GPI, autoimmune and infectious aPL bind to CL in a bilayer, inducing liposome leakage; (2) the leakage mechanism induced by aPL is suggested to be intravesicular; (3) beta2GPI requirement for phospholipid binding in both solid and fluid phase is associated to aPL avidity; (4) CL alone or the CL-beta2GPI complex are the most likely epitopes for autoimmune aPL; (5) aPL from syphilis patients can only form the CL-aPL complex, supporting that beta2GPI is not (part of) the target epitope.


[Parosteal osteosarcomas: unusual findings].
To study and evaluate the unusual findings in the natural history of parosteal osteosarcomas. Parosteal osteosarcomas are well-differentiated tumours of low grade malignancy, but may dedifferentiate into a more aggressive lesion. We reviewed 7 parosteal osteosarcomas treated in La Paz Hospital between 2005 and 2009 (3 were dedifferentiated parosteal osteosarcomas). The authors analysed the clinical and radiological features, histology, treatment and outcomes in this patient group. There were 5 men and 2 women, with a mean age of 32.14 years, range 24-47 years. Three of seven tumours (42.8%) were dedifferentiated osteosarcomas. All three patients received chemotherapy. One patient presented with pulmonary metastases. No patient died. It is important to understand that the biological phenomenon of dedifferentiation can occur in parosteal osteosarcomas. The prognosis, treatment and survival changes radically.


Structural instability caused by a mutation at a conserved arginine in the alpha-crystallin domain of Chinese hamster heat shock protein 27.
Mutations in the alpha-crystallin domain of 4 of the small heat shock proteins (sHsp) (Hsp27/HspB1, alphaA-crystallin/ HspB4, alphaB-crystallin/HspB5, and HspB8) are responsible for dominant inherited diseases in humans. One such mutation at a highly conserved arginine residue was shown to cause major conformational defects and intracellular aggregation of alphaA- and alphaB-crystallins and HspB8. Here, we studied the effect of this Arg mutation on the structure and function of Hsp27. Chinese hamster Hsp27 with Arg148 replaced by Gly (Hsp27R148G) formed dimers in vitro and in vivo, which contrasted with the 12- or 24-subunit oligomers formed by the wild-type protein (Hsp27WT). Despite these alterations, Hsp27R148G had a chaperone activity almost as high as Hsp27WT. The dimers of Hsp27R148G did not further deoligomerize on phosphorylation and like the dimers formed by phosphorylated Hsp27WT were not affected by the deletion of the N-terminal WD/EPF (single letter amino acid code) motif, suggesting that mutation of Arg148, deletion of the N-terminal WD/EPF motif, and phosphorylation of Ser90 may produce similar structural perturbations. Nevertheless, the structure of Hsp27R148G appeared unstable, and the mutated protein accumulated as aggregates in many cells. Both a lower basal level of phosphorylation of Hsp27R148G and the coexpression of Hsp27WT could reduce the frequency of formation of these aggregates, suggesting possible mechanisms regulating the onset of the sHsp-mediated inherited diseases.


Preservation of KIT genotype in a novel pair of patient-derived orthotopic xenograft mouse models of metastatic pediatric CNS germinoma.
Metastatic intracranial germinoma is difficult to treat. Although the proto-oncogene KIT is recognized as one of the most frequent genetic abnormalities in CNS germinoma, the development of new target therapeutic agents for CNS germinoma is hampered by the lack of clinically-relevant animal models that replicate the mutated or over-expressed KIT. CNS germinoma tumor cells from five pediatric patients were directly implanted into the brains of Rag2/severe combined immune deficiency mice. Once established, the xenograft tumors were sub-transplanted in vivo in mouse brains. Characterization of xenograft tumors were performed through histologic and immunohistochemical staining, and KIT mutation analysed with quantitative pyro-sequencing. Expression of putative cancer stem cell markers (CD133, CD15, CD24, CD44, CD49f) was analyzed through flow cytometry. Two patient-derived orthotopic xenograft (PDOX) models (IC-6999GCT and IC-9302GCT) were established from metastatic germinoma and serially sub-transplanted five times in mouse brains. Similar to the original patient tumors, they both exhibited faint expression (+) of PLAP, no expression (-) of β-HCG and strong (+++) expression of KIT. KIT mutation (D816H), however, was only found in IC-9320GCT. This mutation was maintained during the five in vivo tumor passages with an increased mutant allele frequency compared to the patient tumor. Expression of putative cancer stem cell markers CD49f and CD15 was also detected in a small population of tumor cells in both models. This new pair of PDOX models replicated the key biological features of pediatric intracranial germinoma and should facilitate the biological and pre-clinical studies for metastatic intracranial germinomas.


Novel hexarelin analogs stimulate feeding in the rat through a mechanism not involving growth hormone release.
Growth hormone-releasing peptides (GHRPs) are a class of small peptides that stimulate growth hormone (GH) release in several animal species, including the human. Moreover, GHRPs injected into the brain ventricles stimulate feeding in the rat. The aim of this study was to evaluate the GH-releasing properties of a series of novel GHRP analogs and the possible existence of functional correlations between the GH-releasing activity and the effects on feeding behavior. Two well-known hexapeptides, GHRP-6 and hexarelin, given s.c., dose dependently stimulated both GH release and feeding behavior in satiated rats. However, in a series of tri-, penta- and hexapeptide analogs of hexarelin, some compounds were active either on GH release or on eating behavior. Interestingly, even minor structural modifications resulted in major changes of the pharmacological profile. We conclude that GHRPs have orexigenic properties after systemic administration which are largely independent from the effects they exert on GH release.


HLX22, an anti-HER-2 monoclonal antibody, in patients with advanced solid tumors overexpressing human epidermal growth factor receptor 2: an open-label, dose-escalation, phase 1 trial.
HLX22 is a novel monoclonal antibody targeting human epidermal growth factor receptor 2 (HER2). This first-in-human, phase 1 dose-escalation study aimed to evaluate the safety, pharmacokinetics, pharmacodynamics, and preliminary efficacy of HLX22 in patients with advanced solid tumors who had failed or were intolerant to standard therapies. Enrolled patients aged 18 to 75 years with histologically confirmed HER2-overexpressing advanced or metastatic solid tumors received intravenous HLX22 once every 3 weeks at 3, 10, and 25 mg/kg. Primary endpoints were safety and the maximum tolerated dose (MTD). Secondary endpoints included pharmacokinetics, pharmacodynamics, immunogenicity, and efficacy. Between July 31, 2019, and December 27, 2021, 11 patients were enrolled to receive HLX22 at 3 (n = 5), 10 (n = 3), and 25 (n = 3) mg/kg doses. The most common treatment-emergent adverse events were lymphocyte count decreased (45.5%), white blood cell count decreased (36.4%), and hypokalemia (36.4%). No serious adverse events or dose-limiting toxicities occurred during the treatment period, and the MTD was determined at 25 mg/kg once every 3 weeks. Systemic exposure of HLX22 increased with escalating dose levels. No patients achieved a complete or partial response, and four (36.4%) had stable disease. The disease control rate and median progression-free survival were 36.4% (95% confidence interval [CI], 7.9-64.8) and 44.0 days (95% CI, 41.0-170.0), respectively. HLX22 was well tolerated in patients with advanced solid tumors overexpressing HER2 after failure of standard therapies. The study results support further investigation of HLX22 in combination with trastuzumab and chemotherapy.


Utilizing temperature-sensitive association of Pluronic F-127 with lipid bilayers to control liposome-cell adhesion.
The temperature sensitive properties of Pluronic F-127 (MW approximately 12600, PEO(98)-PPO(67)-PEO(98)), a block co-polymer or poloxamer, was used to control liposome-cell adhesion. When associated with liposomes, the PEO moiety of the block co-polymer is expected to inhibit liposome-cell adhesion. Liposomes were made using egg phosphatidylcholine and different mole% of Pluronic F-127. Size measurement of the liposomes at different temperatures, in the presence and absence of Pluronic F-127, shows significant reduction in the size of multilamellar vesicles, at higher temperatures, by the Pluronic molecules. Negative stain electron microscopy study showed the presence of individual molecules and micelles of Pluronic, respectively at temperatures below and above the critical micellar temperature (CMT). Measurement of the surface associated Pluronics indicated that they associated with liposomes when the sample was heated above the Pluronic CMT, and dissociated from liposomes when cooled below the CMT. Attachment of the Pluronic containing liposomes to CHO cells was inhibited at temperatures above the CMT, but not at temperatures below CMT, indicating that temperature-sensitive control of liposome-cell adhesion is achieved.