Influence of neurological lesion level on heart rate variability and fatigue in adults with spinal cord injury.
Group cohort design. To determine the influence of spinal cord injury (SCI) and neurological level on heart rate variability (HRV) and associations with fatigue. SCI rehabilitation outpatient and community settings in New South Wales, Australia. Participants included 45 adults with SCI living in the community and 44 able-bodied controls. Socio-demographic, neurological injury, psychological, HRV and eye blink variables were assessed. Multivariate analysis of variance and post hoc protected t-tests were used to determine differences in HRV and fatigue as a function of the neurological level. Pearson's correlation analysis was used to determine the relationships between these factors. Participants with SCI had significantly reduced sympathetic activity. Those with tetraplegia had lowered sympathetic activity compared with those with paraplegia and able-bodied controls. Neither were differences in parasympathetic activity found between groups nor were there any significant differences found for the time domain or non-linear domains. Higher levels of fatigue were found in the SCI sample, and participants with tetraplegia had higher fatigue levels compared with those with paraplegia. Fatigued participants were more likely to have altered autonomic function-that is, reduced sympathetic activity. Higher levels of neurological impairment in people with SCI are more likely to result in disordered cardiovascular control involving reduced sympathetic activity, whereas elevated fatigue was found to be associated with increased sympathetic dysfunction. Findings highlight the need to address risks associated with this dysfunction, such as improved HRV and fatigue screening for people with SCI and improved education on cardiovascular risk factors.


Testicular teratoma with a unique mode of spread and spontaneous maturation.
Chylous ascites, a rare compliation following retroperitoneal lymph node dissection, has not been reported as a means of spread of testicular cancer. This report describes a unique path of spread of testicular cancer which also appears to spontaneously mature at the metastatic site.


A comparative analysis of Ki-67, p53, and p21WAF1CIP1 expression in tamoxifen associated endometrial carcinomas.
To determine clinicopathologic parameters, expression of proliferation markers, and immunohistochemical oncogene expression in endometrial cancers in patients with a history of breast cancer with and without tamoxifen use. Thirty endometrial carcinoma specimens were examined from patients with a previous history of breast cancer. Patients who had taken tamoxifen (15) were compared to non-users (15). Immunohistochemical staining was performed for p53, Ki-67, and p21WAF1/CIP1, overexpression was defined as greater than 10% positivity. Patient populations were statistically similar. P53 was overexpressed in 73% of tamoxifen users compared to 53% of non-users. Ki-67 was overexpressed in over 90% of user and non-user specimens. p21WAF1/CIP1 was overexpressed in 33% of users and 47% of non-users. Tamoxifen users had shorter time to diagnosis of endometrial cancer than non-users. In this small study, tamoxifen associated tumors expressed p53 more frequently than non-users, while the opposite was observed with p21WAF1/CIP1. This suggests that p53 mutations might play a role in development of tamoxifen associated tumors.


Antibacterial activity, bio-compatibility and osteogenic differentiation of graphene oxide coating on 3D-network poly-ether-ether-ketone for orthopaedic implants.
Poly-ether-ether-ketone (PEEK) has attracted increasing attention as a promising orthopaedic implant material owing to its excellent mechanical properties and biocompatibility. However, its antibacterial properties must be improved as an implant material. In this study, PEEK was sulfonated to obtain a porous surface, and graphene oxide (GO) was deposited to form a coating with antibacterial activity and biocompatibility. After PEEK was sulfonated for different durations, GO was deposited on the surface to prepare the coating (SPEEK-GO), which was then characterised using scanning electron microscopy (SEM), Raman spectroscopy, and contact angle measurements. The in vitro study included antimicrobial and cellular tests. The results showed that the PEEK sulfonated using a 10-min treatment exhibited a uniform porous structure and provided a better basal surface for the deposition of GO. The SPEEK-GO coating displayed strong antibacterial activity against two common dental pathogens. It exhibited good adhesion and proliferation of MC3T3-E1. Moreover, it showed osteogenic differentiation as bone implant material.


Acute bilateral posterior dislocation of the shoulder: one-stage reconstruction of both humeral heads with cancellous autograft and cartilage preservation.
We present a case of bilateral posterior shoulder dislocation after an epileptic seizure. The anterior humeral-head impression fracture was 60% of the articular surface on the right shoulder and 30% on the left shoulder. We performed an early one-stage reconstruction of both humeral heads. A cancellous autograft was used on the left side and an iliac cortico-cancellous autograft on the right side, with preservation of the patient's cartilage. Three years later, the clinical and morphological results were excellent. The discussion focuses on surgical options that range from conservative treatment with excision of the damaged cartilage to immediate hemiarthroplasty. This case is original because of the preservation of the patient's cartilage during reconstruction.


Nanog, Stat-3, and Sox-5 involvement in human fetal temporomandibular joint late development.
The temporomandibular joint (TMJ) is a synovial joint that allows the complex movements essential for life. It connects the jawbone to the skull, working as a sliding hinge. Moreover, pluripotent stem cells are a source of precursors and tissue-specific cells in developing organisms, however, their biodistribution in developing fetal tissues is weakly studied. The aim of our study was analyse immunohistochemical expression of Nanog, Oct-4, Sox-2 and Stat-3 and Sox-5, in TMJ tissue samples from human fetuses aged between the 12th and 20th weeks of intrauterine life. We fixed and processed TMJ tissue samples from human fetuses, histological sections and immunohistochemical procedures were carried out. TMJ histological studies examination did not reveal any difference in the tissue organization between the samples in the studied periods. Immunohistochemical analysis demonstrated that Oct-4 and Sox-2 lack their expression in TMJ. In contrast, Nanog was expressed in nucleous of proliferative layer of mandibular condyle, Stat-3 was expressed in nuclear cells of articular disc, Stat-3 and Sox-5 showed positive nuclear and cytoplasmic immunostaining in codrocyte layers and in ossification areas. Nanog acts in maintanence of pluripotency, Stat-3 in articular disc acts as a transcriptional factor. Stat-3 and Sox-2 act in chondrocyte and osteoblast diferentiation. Distribution of the cells, which express Nanog, Stat-3, and Sox-5 in TMJ tissue during fetal development, can help further understand its physiology, pathology, and repairing capacities.


Menstrual hygiene among adolescent girls studying in a university of Gujarat.
Menstruation marks the beginning of the reproductive life of a girl. A poor menstrual understanding and its unhealthy practices may increase the risk of reproductive infections, urinary tract infections, and even cancer. Therefore, the main objective of this study was studying the knowledge and attitude regarding menstrual hygiene among adolescent girls so that it can be then taken as a background for designing necessary interventions in the community. 1) To study the knowledge and attitude towards menstruation among adolescent girls and 2) to study the menstrual hygiene practices and health-seeking behaviour of the adolescent girls. This was a university-based descriptive cross-sectional study carried out by directly interviewing the subjects using the interviewer-administered questionnaire - pre-tested and pre-designed proforma. The data were analysed statistically by simple proportions. Among the study subjects, 79% had proper knowledge about menstruation. 82% had a normal menstrual pattern. Nearly 96% used sanitary pads, and 2.36% used reusable cloths. Regarding restrictions, 74.80% restrained themselves from visiting the places of worship, 21% avoided physical exercise, and 7.87% had to remain isolated with minimal social contact. Nearly 19% made conscious efforts for dietary changes during menstruation. A majority of the subjects had a regular menstrual cycle, but menstrual problems were found to be more among those having irregular periods, those changing absorbents infrequently, with an inadequate frequency of cleaning, and those using plain water for cleaning. The study however does reflect the fact that menstrual hygiene was unsatisfactory among adolescent girls. Therefore, they need to be educated about the facts of menstruation and proper hygienic practices.


Chair-Based Exercise Interventions for Nursing Home Residents: A Systematic Review.
Despite mobility impairments, many residents have physical, cognitive, and psychosocial resources that should be promoted. The aim was to summarize the current evidence on chair-based exercise (CBE) interventions for nursing home residents. Systematic review registered with Prospero (registration number: CRD42018078196). Nursing home residents in long-term care. Five electronic databases were searched (MEDLINE, Embase, CINAHL, Cochrane Central, and PsycINFO) from inception until July 2020. Title, abstract, and full-text screening as well as quality assessment with the Downs and Black checklist was done by 2 independent reviewers. Studies were eligible if they (1) were conducted in nursing home residents, (2) included participants with a mean age of 65 years, (3) had at least 1 treatment arm with seated exercises only, (4) included active or inactive controls, (5) measured outcomes related to physical and/or cognitive functioning and/or well-being, and (6) controlled studies or single-group pre-post design. Because of a heterogeneity in characteristics of included studies, we refrained from conducting a meta-analysis. Ten studies met the inclusion criteria (n = 511, mean age 79 ± 7 years, 65% female). Studies differed in sample size (12-114) as well as in training type (multicomponent, Yoga/Qigong/breathing exercise, range of motion) and dose (frequency 2 sessions/week to daily, intensity low to moderate, time 20-60 minutes/session, 6 weeks to 6 months). Overall, CBE appears to be feasible and safe. Studies found task-specific improvements in physical and cognitive functions and enhanced well-being. Three studies demonstrated improved lower body performance following a multicomponent CBE program in mobile residents. Three studies only including residents unable to walk reported improved physical functions, indicating that immobile residents benefit from CBE programs. There was a lack of separating mobile and immobile residents in analyses. The results indicate that CBE interventions may improve physical and cognitive functions as well as well-being in nursing home residents. Task-specific multicomponent CBE appears to be best for improving different domains of physical and cognitive functioning. More high-quality trials are needed.


Hodgkin's disease: evaluation of staging procedures for abdominal involvement.
Results of preoperative assessment procedures and subsequent staging laparotomy are evaluated in 32 consecutive patients who presented with extra-abdominal Hodgkin's disease which was not already known to be in an advanced stage. Disease was found in the abdominal cavity of 10 of the 27 patients who presented with disease above the diaphragm, and of two of the five patients in whom disease presented in the inguinal nodes. In patients who initially presented with disease above the diaphragm, the spleen was involved in all 10 cases where abdominal disease was detected. Lymphography was not found to be reliable as a diagnostic aid in doubtful cases. Neither liver nor spleen scans were found to be helpful in assessment of these patients.


Evaluation of the larval migration inhibition assay for detecting macrocyclic lactone resistance in Dirofilaria immitis.
Anthelmintics of the macrocyclic lactone (ML) drug class are widely used as preventives against the canine heartworm (Dirofilaria immitis). Over the past several years, however, reports of ML lack of efficacy (LOE) have emerged, in which dogs develop mature heartworm infection despite the administration of monthly prophylactics. More recently, isolates from LOE cases have been used to infect laboratory dogs and the resistant phenotype has been confirmed by the establishment of adult worms in the face of ML treatment at normally preventive dosages. Testing for and monitoring resistance in D. immitis requires a validated biological or molecular diagnostic assay. In this study, we assessed a larval migration inhibition assay (LMIA) that we previously optimized for use with D. immitis third-stage larvae (L3). We used this assay to measure the in vitro ML susceptibilities of a known-susceptible laboratory strain of D. immitis and three highly suspected ML-resistant isolates originating from three separate LOE cases; progeny from two of these isolates have been confirmed ML-resistant by treatment of an infected dog in a controlled setting. A nonlinear regression model was fit to the dose-response data, from which IC50 values were calculated. The D. immitis LMIA yielded consistent and reproducible dose-response data; however, no statistically significant differences in drug susceptibility were observed between control and LOE parasites. Additionally, the drug concentrations needed to paralyze the L3 were much higher than those third- and fourth-stage larvae would experience in vivo. IC50 values ranged from 1.57 to 5.56μM (p≥0.19). These data could suggest that ML resistance in this parasite is not mediated through a reduced susceptibility of L3 to the paralytic effects of ML drugs, and therefore motility-based assays are likely not appropriate for measuring the effects of MLs against D. immitis in this target stage.


GSK-3beta-dependent destabilization of cyclin D1 mediates replicational stress-induced arrest of cell cycle.
Chemotherapeutic agents are well known to induce growth arrest of cancerous cells by inducing DNA damage/replicational stress and engaging cellular apoptotic machinery. Our studies on hydroxyurea (HU) recognized cyclin D1 destabilization as the initiator of growth arrest at G(1)/S-phase independent of other cell cycle regulators. Cyclin D1 degradation was associated with its phosphorylation at Thr286 by glycogen synthase kinase-3beta and inactivation of Akt kinase. Overexpression of the cyclin D1(T286A) mutant, or constitutively active Akt, conferred stability to cyclin D1 and helped bypass cell cycle arrest. Thus, growth arrest by HU seems to involve destabilization of cyclin D1 in addition to its well-established role as ribonucleotide reductase inhibitor.


Exploring the Efficacy of a Set of Smart Devices for Postural Awareness for Workers in an Industrial Context: Protocol for a Single-Subject Experimental Design.
In manufacturing industries, tasks requiring poor posture, high repetition, and long duration commonly induce fatigue and lead to an increased risk of work-related musculoskeletal disorders. Smart devices assessing biomechanics and providing feedback to the worker for correction may be a successful way to increase postural awareness, reducing fatigue, and work-related musculoskeletal disorders. However, evidence in industrial settings is lacking. This study protocol aims to explore the efficacy of a set of smart devices to detect malposture and increase postural awareness, reducing fatigue, and musculoskeletal disorders. A longitudinal single-subject experimental design following the ABAB sequence will be developed in a manufacturing industry real context with 5 workers. A repetitive task of screw tightening of 5 screws in a standing position into a piece placed horizontally was selected. Workers will be assessed in 4 moments per shift (10 minutes after the beginning of the shift, 10 minutes before and after the break, and 10 minutes before the end of the shift) in 5 nonconsecutive days. The primary outcomes are fatigue, assessed by electromyography, and musculoskeletal symptoms assessed by the Nordic Musculoskeletal Questionnaire. Secondary outcomes include perceived effort (Borg perceived exertion scale); range of motion of the main joints in the upper body, speed, acceleration, and deceleration assessed by motion analysis; risk stratification of range of motion; and cycle duration in minutes. Structured visual analysis techniques will be conducted to observe the effects of the intervention. Results for each variable of interest will be compared among the different time points of the work shift and longitudinally considering each assessment day as a time point. Enrollment for the study will start in April 2023. Results are expected to be available still in the first semester of 2023. It is expected that the use of the smart system will reduce malposture, fatigue, and consequently, work-related musculoskeletal pain and disorders. This proposed study will explore a strategy to increase postural awareness in industrial manufacturing workers who do repetitive tasks, using smart wearables that provide real-time feedback about biomechanics. Results would showcase a novel approach for improving self-awareness of risk for work-related musculoskeletal disorders for these workers providing an evidence base support for the use of such devices. PRR1-10.2196/43637.


Dietary whey protein downregulates fatty acid synthesis in the liver, but upregulates it in skeletal muscle of exercise-trained rats.
This study compared the effects of casein and whey protein as the source of dietary protein on the activity of lipogenic enzymes and mRNA levels in the liver and skeletal muscle of exercise-trained rats. Twenty-eight male Sprague-Dawley rats were randomly assigned to one of four groups (n = 7/group). Rats were assigned to sedentary or exercise-trained groups and were fed the casein or whey protein diet. Rats in the exercise groups were trained for 2 wk using a swimming exercise for 120 min/d and 6 d/wk. A significant decrease in the activity of the hepatic lipogenic enzymes, glucose-6-phosphate dehydrogenase, malic enzyme, adenosine triphosphate citrate lyase, acetyl-coenzyme A carboxylase, and fatty acid synthase (FASN) was observed in rats fed whey protein compared with animals fed casein. Compared with the casein diet, the whey protein diet also lowered mRNA expression of these enzymes, except for FASN. In contrast to the findings in liver, whey protein, as compared with casein, increased skeletal muscle FASN activity and mRNA. Further, exercise training resulted in increased skeletal muscle glucose-6-phosphate dehydrogenase and FASN activity and adenosine triphosphate citrate lyase, acetyl-coenzyme A carboxylase-1, and FASN mRNA expression. Exercise training or whey protein may play an important role in suppressing hepatic fatty acid synthesis, thereby decreasing accumulation of body fat and stimulating the skeletal muscle to increase energy substrate as fat during prolonged exercise.


Effects of epidural block anesthesia combined with general anesthesia on inflammatory factors, cognitive function and postoperative pain in patients with lung cancer after thoracoscopic surgery.
To investigate the effects of epidural block anesthesia combined with general anesthesia on inflammatory factors, cognitive function and postoperative pain in patients with lung cancer after thoracoscopic surgery. A total of 144 lung cancer patients admitted to the Department of Cardiothoracic Surgery of Ganzi Tibetan Autonomous Prefecture People's Hospital from October 2017 to October 2019 were included in this retrospective cohort study. The patients were divided into an observation group and a control group, with 72 cases in each group. Observation group was treated with epidural block anesthesia plus general anesthesia under thoracoscopic surgery, while control group was treated with general anesthesia alone. General information of both groups was compared. Tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels were measured by enzyme-linked immunosorbent assay before anesthesia (T0), before the end of operation (T1), 12 h after operation (T2) and 24 h after operation (T3). Calcium-binding protein (S-100β) content, mini-mental state examination (MMSE) and Montreal cognitive assessment (MoCA) at T0, T3, the 3rd day after operation (T4), and the 5th day after operation (T5) were measured. And postoperative pain was recorded. There were no significant differences in TNF-α, IL-6, MMSE and MoCA between the two groups at T0, and no significant differences were seen in S-100β between the two groups at T0 and T1 (all P>0.05). Compared with control group, observation group had lower TNF-α and IL-6 at T1, T2, T3, T4 and T5, and lower S-100β at T3, T4 and T5 (all P<0.001). Lower pain scores and higher MMSE and MoCA were found in the observation group at T3, T4 and T5 (all P<0.05). Epidural block anesthesia combined with general anesthesia can effectively reduce levels of inflammatory factors, cognitive disorder and postoperative pain in patients with lung cancer after thoracoscopic surgery.


Hematological and biochemical effects of sub-chronic artesunate exposure in rats.
Artesunate is a potent and rapidly acting blood schizontocide used to treat chloroquine resistant malaria. Artesunate has been reported to cause embryo, reduced reproductive capacity, hepatotoxicity, neurotoxicity and hematological abnormalities. Previously toxicity studies on artesunate have been done in 2-10 mg/kg dose range mostly for 7 days, scientific studies on sub-chronic exposure of artesunate is not been reported so for. The present study evaluates sub-chronic safety profile of artesunate on 45 days oral administration at 2, 4 and 8 mg/kg/day. Authentication of artesunate has been done by color test, pH, melting point, loss on drying, UVmax, TLC and HPLC study. Artesunate has non-significant effect on liver and kidney weight. Serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), serum alkaline phosphate (ALP), cholesterol (TC), triglyceride (TG), total protein, albumin, bilirubin, creatinine, urea and glucose content were estimated after 45 days treatment along with hematological screening. Artesunate treatment for 45 days significantly increased (p < 0.05-0.001) SGOT, SGPT, ALP, TC, TG, total bilirubin, glucose level at 8 mg/kg/day dose. It has non-significant effect on serum total protein, albumin, creatinine and urea. Hemoglobin, total RBC, platelet, lymphocytes, basophil, mean cell volume and mean corpuscular hemoglobin concentration have not changed but total WBC, neutrophil, eosinophil, packed cell volume and mean cell hemoglobin were increased significantly (p < 0.01) at 8 mg/kg/day dose. Artesunate treatment at 4 and 8 mg/kg/day showed sinusoidal dilation, cytoplasmic vaculation, focal necrosis, sinusoidal congestion and extensive inflammatory changes, whereas kidney was free of any deleterious effect. Sub-chronic exposure of artesunate at 8 mg/kg/day dose for 45 days period cause hepatic damage along with hematological abnormalities signifying safety concern.


Development of an electronic database for quality assurance of radiotherapy in the International Society of Paediatric Oncology (Europe) high risk neuroblastoma study.
Quality assurance of radiotherapy is an important determinant of outcome in some cancers. SIOPEN-R-NET developed a computerised remote data entry system for recording imaging and treatment parameters for its multimodality high risk neuroblastoma study. This will enable investigation of the relationship between radiotherapy quality and local control.


Risk factors for revision due to prosthetic joint infection following total knee arthroplasty based on 62,087 knees in the Finnish Arthroplasty Register from 2014 to 2020.
Periprosthetic joint infection (PJI) is the commonest reason for revision after total knee arthroplasty (TKA). We assessed the risk factors for revision due to PJI following TKA based on the Finnish Arthroplasty Register (FAR). We analyzed 62,087 primary condylar TKAs registered between June 2014 and February 2020 with revision for PJI as the endpoint. Cox proportional hazards regression was used to estimate hazard ratios (HR) with 95% confidence intervals (CI) for the first PJI revision using 25 potential patient- and surgical-related risk factors as covariates. 484 knees were revised for the first time during the first postoperative year because of PJI. The HRs for revision due to PJI in unadjusted analysis were 0.5 (0.4-0.6) for female sex, 0.7 (0.6-1.0) for BMI 25-29, and 1.6 (1.1-2.5) for BMI > 40 compared with BMI < 25, 4.0 (1.3-12) for preoperative fracture diagnosis compared with osteoarthritis, and 0.7 (0.5-0.9) for use of an antimicrobial incise drape. In adjusted analysis the HRs were 2.2 (1.4-3.5) for ASA class III-IV compared with class I, 1.7 (1.4-2.1) for intraoperative bleeding ≥ 100 mL, 1.4 (1.2-1.8) for use of a drain, 0.7 (0.5-1.0) for short duration of operation of 45-59 minutes, and 1.7 (1.3-2.3) for long operation duration > 120 min compared with 60-89 minutes, and 1.3 (1.0-1.8) for use of general anesthesia. We found increased risk for revision due to PJI when no incise drape was used. The use of drainage also increased the risk. Specializing in performing TKA reduces operative time and thereby also the PJI rate.


Targeting lactate dehydrogenase A (LDHA) exerts antileukemic effects on T-cell acute lymphoblastic leukemia.
T-cell acute lymphoblastic leukemia (T-ALL) is an uncommon and aggressive subtype of acute lymphoblastic leukemia (ALL). In the serum of T-ALL patients, the activity of lactate dehydrogenase A (LDHA) is increased. We proposed that targeting LDHA may be a potential strategy to improve T-ALL outcomes. The current study was conducted to investigate the antileukemic effect of LDHA gene-targeting treatment on T-ALL and the underlying molecular mechanism. Primary T-ALL cell lines Jurkat and DU528 were treated with the LDH inhibitor oxamate. MTT, colony formation, apoptosis, and cell cycle assays were performed to investigate the effects of oxamate on T-ALL cells. Quantitative real-time PCR (qPCR) and Western blotting analyses were applied to determine the related signaling pathways. A mitochondrial reactive oxygen species (ROS) assay was performed to evaluate ROS production after T-ALL cells were treated with oxamate. A T-ALL transgenic zebrafish model with LDHA gene knockdown was established using CRISPR/Cas9 gene-editing technology, and then TUNEL, Western blotting, and T-ALL tumor progression analyses were conducted to investigate the effects of LDHA gene knockdown on T-ALL transgenic zebrafish. Oxamate significantly inhibited proliferation and induced apoptosis of Jurkat and DU528 cells. It also arrested Jurkat and DU528 cells in G0/G1 phase and stimulated ROS production (all P < 0.001). Blocking LDHA significantly decreased the gene and protein expression of c-Myc, as well as the levels of phosphorylated serine/threonine kinase (AKT) and glycogen synthase kinase 3 beta (GSK-3β) in the phosphatidylinositol 3'-kinase (PI3K) signaling pathway. LDHA gene knockdown delayed disease progression and down-regulated c-Myc mRNA and protein expression in T-ALL transgenic zebrafish. Targeting LDHA exerted an antileukemic effect on T-ALL, representing a potential strategy for T-ALL treatment.


Automatic segmentation of human brain sulci.
The neocortical surface has a rich and complex structure comprised of folds (gyri) and fissures (sulci). Sulci are important macroscopic landmarks for orientation on the cortex. A precise segmentation and labeling of sulci is helpful in human brain mapping studies relating brain anatomy and function. Due to their structural complexity and inter-subject variability, this is considered as a non-trivial task. An automatic algorithm is proposed to accurately segment neocortical sulci: vertices of a white/gray matter interface mesh are classified under a Bayesian framework as belonging to gyral and sulcal compartments using information about their geodesic depth and local curvature. Then, vertices are collected into sulcal regions by a watershed-like growing method. Experimental results demonstrate that the method is accurate and robust.


Preparation of bifunctional monomer molecularly imprinted polymer filled solid-phase extraction for sensitivity improvement of quantitative analysis of sulfonamide in milk.
To solve the problems of unstable adsorption performance and high organic solvent consumption of traditional molecular imprinting materials, we developed a water-resistant and highly adsorptive molecularly imprinted sulfamethazine polymer (MIP) through a novel synthetic strategy consisting of the application of mixed functional monomers combined with hydrophobic crosslinkers. The results showed that the imprinting factor of the prepared MIP was increased from 0.93 to 4.38, and that the relative selection coefficient k' was 5.59. With the application of the material to be developed as a solid-phase extraction (SPE) filler for sulfonamides (SAs) in milk combined with UPLC-MS/MS, the validated method showed a sensitive quantification limit (0.89 µg/kg), a steady recovery (95.55%-97.97%) and an excellent precision (0.08%-2.92% RSD). Moreover, after 5 times usage, the recovery rate of MIP-SPE was still above 90%. Therefore, the prepared materials showed high performance of molecular adsorption and were water-resistant, which was also considered an excellent filler of SPE for SAs extraction in food or other fields.


Benefit-cost analysis of becoming certified pollinator friendly.
Despite significant drop in pollinator abundance, no studies exist on the benefits and costs of pollinator conservation in the public domain. An in-person survey was conducted at three large, public US universities to estimate benefits to become Bee Campus USA certified. We test whether different types of reminders on existing student sustainability fees affect Willingness to Pay. Costs of achieving this certification per university were obtained. Net Present Value demonstrates that the net benefits to each school are largely positive, except under the most restrictive assumption. Information reminders of existing fees lead to little change in support of pollinator conservation.


Assessment of elastin maturation by radioimmunoassay of desmosine in the developing human lung.
Desmosine has been quantitated in the normally grown fetal and early infant lung by radioimmunoassay. Desmosine could first be detected at 22 weeks gestation: the concentration of desmosine expressed per milligram lung DNA increased in approximately linear form up to about 55 weeks postconceptional age. The concentration in peripheral lung was approximately half that in whole lung homogenates. Lungs of infants dying with acute HMD and lungs of growth retarded infants showed no significant differences from the normals, although there was a tendency for higher desmosine concentrations in prematurely born growth retarded infants.


Detection of tau-fluvalinate resistance in the mite Varroa destructor based on the comparison of vial test and PCR-RFLP of kdr mutation in sodium channel gene.
Varroa destructor is the major cause of honey bee (Apis mellifera) colony losses. Mite control is limited to several miticides. The overuse of tau-fluvalinate has resulted in resistance via a knockdown resistance (kdr) mutation in the sodium channel gene NaVChs (L925V/I/M). In this study, we used the discriminating concentration of tau-fluvalinate (0.25 µg/mL) to detect the resistance of mites in a bioassay. Further, we verified the presence of the kdr mutation in mites from the bioassay via PCR amplification of a fragment of the voltage-gated sodium channel gene (NaVCh), restriction fragment length polymorphisms (RFLPs), and densitometry analyses in pools of surviving or dead mites. Resistance values corresponding to the densitometry of the resistant allele were related to mite survival. In the vial test, the survival of the control group was significantly higher (70.4%) than that of the tau-fluvalinate-treated group (34.3%). Mite survival in the vial test was significantly correlated with the mean proportion of resistance values. Individuals that died after tau-fluvalinate application exhibited an average resistance value of 0.0783, whereas individuals that survived exhibited an average resistance of 0.400. The concentration of tau-fluvalinate in the vials was checked using high performance liquid chromatography under different temperatures and exposure times, and indicates that the stability of tau-fluvalinate stored in the refrigerator (4 ± 1 °C) is at least 14 days. PCR-RFLP of the NaVCh gene fragment verified that the vial test is a suitable, rapid, and cost-effective method for the identification of tau-fluvalinate resistance based on kdr mutation in V. destructor in apiaries.


IL-3-dependent early erythropoiesis is stimulated by autocrine transforming growth factor beta.
Autocrine/paracrine transforming growth factor beta (TGF-beta) is an important regulator of stem cell quiescence and generally suppresses stem cell proliferation. However, we show here that during the first few days of an erythroid cell culture from adult blood stem cells, the presence of neutralizing antibodies against TGF-beta had a suppressive effect on subsequent erythropoiesis, indicating a stimulatory action of autocrine TGF-beta. The suppression occured in the form of a delay in erythroblast proliferation rather than a reduction in final erythroid colony numbers. The inhibitory effect of anti-TGF-beta occured in the presence of interleukin-3 (IL-3) but not in cultures with only stem cell factor and erythropoietin. Erythroblasts expressing gamma-globin (gamma+) were more strongly suppressed than erythroblasts expressing only beta-globin (gamma-beta+), so that stem cell treatment with anti-TGF-beta caused a decrease in the proportion of gamma+ cells. Anti-TGF-beta had an inhibitory effect on erythropoiesis only when administered during the first 4 days of culture, that is, before the onset of globin expression and dependence on erythropoietin. The decreasing effect of anti-TGF-beta with delayed addition coincided with a decreasing dependence on IL-3. CD133+ stem cells were more strongly suppressed by anti-TGF-beta than the complementary CD133-CD34+ stem cells, and the latter were also much less dependent on IL-3. The treatment of very early stem cell cultures with a pulse of added TGF-beta1 in the presence of IL-3 increased the subsequent proliferation of erythroblasts. Taken together, the data suggest that IL-3-driven early erythropoiesis from immature peripheral blood stem cells is stimulated by autocrine TGF-beta.


AZD4547 and calcitriol synergistically inhibited BT-474 cell proliferation while modified stemness and tumorsphere formation.
Fibroblast growth factor receptor (FGFR) overamplification/activation in cancer leads to increased cell proliferation. AZD4547, a FGFR selective inhibitor, hinders breast cancer cells growth. Although luminal B breast tumors may respond to chemotherapy and endocrine therapy, this subtype is associated with poor prognosis, inadequate response and/or acquired drug resistance. Calcitriol, the vitamin D most active metabolite, exerts anti-neoplastic effects and enhances chemotherapeutic drugs activity. In this study, we sought to decrease the concentration of AZD4547 needed to inhibit the luminal-B breast cancer cell line BT-474 proliferation by its combination with calcitriol. Anti-proliferative inhibitory concentrations, combination index and dose-reduction index were analyzed from Sulforhodamine B assays. Western blot and qPCR were used to study FGFR molecular targets. The compound's ability to inhibit BT-474 cells tumorigenic capacity was assessed by tumorspheres formation. Results: BT-474 cells were dose-dependently growth-inhibited by calcitriol and AZD4547 (IC50 = 2.9 nM and 3.08 μM, respectively). Calcitriol at 1 nM synergistically improved AZD4547 antiproliferative effects, allowing a 2-fold AZD4547 dose-reduction. Mechanistically, AZD4547 downregulated p-FGFR1, p-Akt and tumorsphere formation. Calcitriol also decreased tumorspheres, while induced cell differentiation. Both compounds inhibited MYC and CCND1 expression, as well as ALDH, a stemness marker that positively correlated with FGFR1 and negatively with VDR expression in breast cancer transcriptomic data. In conclusion, the drugs impaired self-aggregation capacity, reduced stemness features, induced cell-differentiation and when combined, synergistically inhibited cell proliferation. Overall, our results suggest that calcitriol, at low pharmacological doses, may be a suitable candidate to synergize AZD4547 effects in luminal B breast tumors, allowing to reduce dose and adverse effects.


The gut microbiota and diabetic cardiomyopathy in humans.
Type 2 diabetes (DT2) increases the risk of cardiovascular events and cardiac insufficiency. This insufficiency is mostly post-ischaemic in nature, but other aetiologies are possible in this high-risk population. In patients with DT2, diabetic cardiomyopathy is a recognized cause of cardiac insufficiency secondary to chronic hyperglycaemia and myocardial lipotoxicity, which promotes cardiomyocyte hypertrophy (and, frequently, apoptosis of these cells), interstitial fibrosis and a decrease in myocardial contractile performance. Several studies have shown that diabetic cardiomyopathy is associated with modifications to the intestinal microbiota, and changes in the synthesis of bacterial metabolites and their diffusion into the host, some of which appear to have direct deleterious effects on cardiac contractility. These findings open up new perspectives for pathophysiological studies by establishing the presence of a 'microbiota-myocardium' axis and raising the possibility of innovative new treatments. Correction of intestinal dysbiosis in patients with cardiac insufficiency could, therefore, constitute an innovative therapeutic approach to cases of this disease with a poor prognosis.


[Diagnosis and treatment of cervical disc calcification in children].
To investigate the diagnosis and treatment cervical disc calcification in children. 13 children with cervical disc calcification were analysed retrospectively. Cervical pain was observed after influenza in 5 children, suddenly cervical pain in 3, sudden wryneck in 3 cases. Two children with this disease were accidentally found after injury. 14 calcifications of cervical disks were found in 13 children. Clinical symptoms disappeared after symptomatic management. Calcification disappeared in 5 months. Cervical disc calcification in children is a kind of benign self-limited disease. Correst understanding of pathophysiologic course of this disease helps to avoid unnecessary surgery.


Clinical associations with uterine tachysystole.
To determine the incidence of uterine tachysystole (UT) and its association with neonatal depression or metabolic acidemia (DEP). This retrospective study comprised all 6234 women at ≥ 37 weeks' gestation who were monitored during the last 4 hours of tracings before birth in an academic community hospital. DEP was defined by an umbilical artery base deficit value ≥ 10 mmol/L or a 5-minute Apgar ≤ 6 and included 77 births. UT was defined by >15 contractions in 30 minutes. The overall incidence of UT was 18.3% (1139/6234). In 4.2% (260/6234) UT persisted for >60 min. The rate of UT was similar in births with DEP (14.3%, 11/77) compared to those without DEP (18.3%, 1128/6157; p=0.45). In births with UT, only 1.0% (11/1139) developed DEP. The DEP group had more decelerations at almost every level of contractions and a higher cesarean rate of 49.4% (38/77) compared to 24.0% (1468/6124); p=<0.001 in the group without DEP. UT was common, occasionally prolonged and almost always benign. Fetuses with DEP had no more UT than those without DEP. Many babies with DEP declared their vulnerability with decelerations at contraction rates below UT levels and the great majority of them never experienced UT.


The Stigma subscale of the Consumer Experiences of Stigma Questionnaire (CESQ): a psychometric evaluation in Polish psychiatric patients.
There is a scarcity of well validated measures of the subjective experience of the stigma of mental illness. The aim of this paper was to evaluate the psychometric properties of the Stigma subscale of the Consumer Experiences of Stigma Questionnaire (CESQ). A secondary analysis of the data from two cross-sectional studies using the CESQ was conducted in order to assess the factor structure, internal consistency and concurrent validity of the Stigma subscale. Sample 1 included 373 diagnostically heterogeneous patients (65% had psychotic disorders) and Sample 2 included 136 patients with schizophrenia. Since none of the factor models of the original 9-item scale was satisfactory, it was shortened by removing two psychometrically weakest items. The abbreviated 7-item scale proved to be a one-dimensional instrument, with good estimates of internal consistency. Its concurrent validity was partly confirmed by demonstrating negative correlation with quality of life and positive correlations with self-rated psychopathological symptoms. However, contrary to theoretical expectations, the stigma score was not associated with global functioning and clinician-rated symptoms. The psychometric properties of the Stigma subscale of the CESQ may be improved by shortening the instrument. Although the reliability and validity of the abbreviated version were partly established, further research is needed to explore in particular its test-retest reliability and concurrent validity.


Effects of resveratrol on the proliferation and osteogenic differentiation of deciduous dental pulp stem cells from neurofibromatosis type 1 patient.
We aimed at verifying whether resveratrol can decrease cell proliferation and change osteogenic differentiation of cells obtained from patients with type 1 neurofibromatosis (NF1). Deciduous dental pulp derived stem cells were isolated from NF1 patient and healthy volunteer. These cells were subjected to increasing concentrations of resveratrol and evaluated for proliferation and mineralization of osteogenic differentiation. The results showed that resveratrol reduced the difference in proliferation between CNT and NF1 cells in a dose-dependent manner and this property was more prominent in affected cells than in healthy cells. Resveratrol showed no statistically significant changes in mineralization in osteogenic differentiation of NF1 cells, at low doses tested. In conclusion, in a dose-dependent manner, resveratrol displays interesting properties that could be applied in a possible treatment aimed at decreasing cellular proliferation in neurofibromatosis. Furthermore, it is selective concerning healthy cells and not affecting cell differentiation. Further research to cell selectivity, differentiation to other tissue types, and cell cytotoxicity are needed.


The influence of green finance availability to retailers on purchase intention: a consumer perspective with the moderating role of consciousness.
As the global warming crisis is increasing daily, it is crucial to find ways to reduce the carbon footprint generated by activities like the production, consumption, and distribution of goods and services. This empirical study has looked at one approach through which environment-friendly production and consumption can be encouraged. The developed model has studied the relationship between retailers' access to green finance and consumer purchase intention of green products by incorporating the role of environmental, status, and future consciousness. Theoretical foundations for this model have been taken from the theory of planned behaviour (TPB) and theory of reasoned action (TRA), which have extensively discussed the role of consciousness and societal norms while making purchase intentions. To gain insights about the purchasing behaviour of consumers, this study collected data from the Jiangsu province of China, where a non-probability convenience sampling technique was used to distribute a questionnaire to 400 respondents between February 2022 and August 2022. The collected data was analysed using Structural Equation Modeling (SEM) in SmartPLS in order to study the relationship between independent and dependent variables. Results of this study show that retailers' access to green finance positively impacts consumer purchase intention towards green products, and adding a consciousness perspective in the model strengthens this relationship. Moreover, the theory of planned behaviour and the theory of reasoned action were validated through this study, providing insights for policymakers on the importance of promoting green finance to influence green product purchase intention. Overall, this study shows that policymakers should give green financing to retailers and environmental and future awareness to consumers to encourage environment-friendly behaviour.


Effect of mercuric chloride on the digestive system of a teleost fish, Channa punctatus.
The effect of 1.8 mg/liter (LC50) of mercuric chloride exposure on the activities of alkaline phosphatase, acid phosphatase, glucose-6-phosphatase, amylase, pepsin, trypsin, tripeptidase glycyl-glycine dipeptidase and carnosinase has been examined in Channa punctatus. The three phosphatases have been inhibited in the liver but showed an increase in activity in the intestine and pyloric caeca. Amylase, pepsin and trypsin have also shown a slight increase in activity. There has been no significant alteration in the activites of the peptidases. The results show that mercury inhibits the activites of phosphatases in the liver but has no significant effect on the digestive enzymes within the experimental period of 96 hours.


Risk factors for lamb mortality on UK sheep farms.
Data on neonatal-lamb mortality and flock history were recorded by veterinary students working on 108 sheep farms in the UK in Spring 1997. The mean reported mortality incidence risk was 10.0% (median 9%; inter-quartile range 5.9-12.3%). The outcomes were incidence risks of stillbirth, perinatal (within 24h of birth) mortality and postnatal (>24h after birth) mortality. Exposures were screened for univariable associations with mortality risk. Logistic binomial multiple regressions adjusted for confounding, with farm as a random effect. This study has raised hypotheses for management factors associated with lamb mortality that warrant further study. Intensive rearing systems appear to be associated with increased perinatal and postnatal mortality, although housing ewes at lambing was associated with a decreased risk of stillbirth. High perinatal mortality also was associated with poor mothering-pen hygiene, flocks that foster more lambs, and failure to provide appropriate nursing for sick lambs. Larger flocks, poor ewe condition at breeding, and flocks with higher ewe-replacement rates were associated with higher postnatal mortality.


Musculoskeletal Pain, Physical Function, and Quality of Life After Bariatric Surgery.
To evaluate the longitudinal effects of metabolic and bariatric surgery (MBS) on the prevalence of musculoskeletal and lower extremity (LE) pain, physical function, and health-related quality of life. The Teen Longitudinal Assessment of Bariatric Surgery study (NCT00474318) prospectively collected data on 242 adolescents undergoing MBS at 5 centers over a 3-year follow-up. Joint pain and physical function outcomes were assessed by using the Health Assessment Questionnaire Disability Index, Impact of Weight on Quality of Life - Kids, and the Short Form 36 Health Survey. Adolescents with Blount disease (n = 9) were excluded. Prevalent musculoskeletal and LE pain were reduced by 40% within 12 months and persisted over 3 years. Adjusted models revealed a 6% lower odds of having musculoskeletal pain (odds ratio = 0.94, 95% confidence interval: 0.92-0.99) and a 10% lower odds of having LE pain (odds ratio = 0.90, 95% confidence interval: 0.86-0.95) per 10% reduction of BMI. The prevalence of poor physical function (Health Assessment Questionnaire Disability Index score >0) declined from 49% to <20% at 6 months (P < .05), Physical comfort and the physical component scores, measured by the Impact of Weight on Quality of Life - Kids and the Short Form 36 Health Survey, improved at 6 months postsurgery and beyond (P < .01). Poor physical function predicted persistent joint pain after MBS. Joint pain, impaired physical function, and impaired health-related quality of life significantly improve after MBS. These benefits in patient-reported outcomes support the use of MBS in adolescents with severe obesity and musculoskeletal pain and suggest that MBS in adolescence may reverse and reduce multiple risk factors for future joint disease.


Nanotribological and Nanomechanical Properties Changes of Tooth After Bleaching and Remineralization in Wet Environment.
Teeth bleaching cases had increased with people's desire for oral aesthetic; however, bleached teeth would still undertake chewing actions and remineralizing process in saliva. Nanotribological and nanomechanical properties are proper displays for dental performance of bleached teeth. The purpose of the research was to reveal the effect of bleaching and remineralization on the nanotribological and nanomechanical properties of teeth in wet environment. The specimens were divided into four groups according to the bleaching products used: 12 % hydrogen peroxide (HP) (12HP group); 15 % carbamide peroxide (CP) (15CP group); 35 % CP (35CP group); and artificial saliva (control group). The nanotribological and nanomechanical property changes of tooth enamel after bleaching and remineralization were evaluated respectively by nanoscratch and nanoindentation tests in wet environment, imitating the wet oral environment. The morphology changes were evaluated by statistical parametric mapping (SPM) and scanning electron microscopy (SEM). After bleaching, 12HP group and 15CP group showed increased scratch depth with more pile ups on the scratch edges, decreased nanohardness, and corroded surface appearance. While the 35CP group showed an increase in nanoscratch depth, no change in nanohardness and surface appearance was observed. The control group showed no change in these measurements. After remineralization, the three bleaching groups showed decreased nanoscratch depth and no change of nanohardness compared with the bleached teeth. And the control group showed no changes in nanotribological and nanomechanical properties. The nanotribological and nanomechanical properties of the 12HP group and 15CP group were affected by bleaching, but the nanotribological properties recovered partly and the nanomechanical properties got no change after 1 week of remineralization. As for the 35CP group, the nanotribological properties were influenced and the nanomechanical properties were not affected. These results remind us of taking actions to protect our teeth during bleaching.


Transition-Metal-Doped RuIr Bifunctional Nanocrystals for Overall Water Splitting in Acidic Environments.
The establishment of electrocatalysts with bifunctionality for efficient oxygen evolution reaction (OER) and hydrogen evolution reaction (HER) in acidic environments is necessary for the development of proton exchange membrane (PEM) water electrolyzers for the production of clean hydrogen fuel. RuIr alloy is considered to be a promising electrocatalyst because of its favorable OER performance and potential for HER. Here, the design of a bifunctional electrocatalyst with greatly boosted water-splitting performance from doping RuIr alloy nanocrystals with transition metals that modify electronic structure and binding strength of reaction intermediates is reported. Significantly, Co-RuIr results in small overpotentials of 235 mV for OER and 14 mV for HER (@ 10 mA cm-2 current density) in 0.1 m HClO4 media. Therefore a cell voltage of just 1.52 V is needed for overall water splitting to produce hydrogen and oxygen. More importantly, for a series of M-RuIr (M = Co, Ni, Fe), the catalytic activity dependence at fundamental level on the chemical/valence states is used to establish a novel composition-activity relationship. This permits new design principles for bifunctional electrocatalysts.


β-Galactosidase from Lactobacillus helveticus DSM 20075: Biochemical Characterization and Recombinant Expression for Applications in Dairy Industry.
β-Galactosidase encoding genes lacLM from Lactobacillus helveticus DSM 20075 were cloned and successfully overexpressed in Escherichia coli and Lactobacillus plantarum using different expression systems. The highest recombinant β-galactosidase activity of ∼26 kU per L of medium was obtained when using an expression system based on the T7 RNA polymerase promoter in E. coli, which is more than 1000-fold or 28-fold higher than the production of native β-galactosidase from L. helveticus DSM 20075 when grown on glucose or lactose, respectively. The overexpression in L. plantarum using lactobacillal food-grade gene expression system resulted in ∼2.3 kU per L of medium, which is approximately 10-fold lower compared to the expression in E. coli. The recombinant β-galactosidase from L. helveticus overexpressed in E. coli was purified to apparent homogeneity and subsequently characterized. The Km and vmax values for lactose and o-nitrophenyl-β-d-galactopyranoside (oNPG) were 15.7 ± 1.3 mM, 11.1 ± 0.2 µmol D-glucose released per min per mg protein, and 1.4 ± 0.3 mM, 476 ± 66 µmol o-nitrophenol released per min per mg protein, respectively. The enzyme was inhibited by high concentrations of oNPG with Ki,s = 3.6 ± 0.8 mM. The optimum pH for hydrolysis of both substrates, lactose and oNPG, is pH 6.5 and optimum temperatures for these reactions are 60 and 55 °C, respectively. The formation of galacto-oligosaccharides (GOS) in discontinuous mode using both crude recombinant enzyme from L. plantarum and purified recombinant enzyme from E. coli revealed high transgalactosylation activity of β-galactosidases from L. helveticus; hence, this enzyme is an interesting candidate for applications in lactose conversion and GOS formation processes.


Short-term safety and efficacy of peroral endoscopic myotomy for the treatment of achalasia in children.
Peroral endoscopic myotomy (POEM) has shown excellent results for the treatment of achalasia in adults, but studies for children are limited. The study was aimed to analyze outcomes of peroral endoscopic myotomy (POEM) in children and compared with those in adults in a large multi-center study. Records of consecutive patients with achalasia who underwent POEM at three tertiary centers were reviewed. A total of 130 children were included in this study. The primary outcomes of perioperative outcomes and clinical follow-up data were analyzed. One child (0.8%) experienced technical failure. Five children (3.8%) had major adverse events, including one with pneumothorax requiring drainage, two with delayed mucosa barrier failure, one with readmission, and one with vital-sign instability. Both post-POEM Eckardt score and median LES pressure were significantly lower than their pre-POEM reference values in children (0.7 vs 7.4; 7.0 vs 27.1 mmHg; both P < 0.001). During a median follow-up time of 40 months, clinical reflux rate was 27.0% and clinical failure rates at 1, 3, and 5 years were 1.8%, 3.5%, and 4.4% for children. The technical failure, major adverse events, and postoperative clinical reflux were comparable between children and adults (all P > 0.05). Kaplan-Meier analysis showed that the risk of clinical failure was lower in children than adults (log-rank test, hazard ratio = 0.37, 95% confidence interval 0.15-0.91, P = 0.023). POEM can be safely performed in children with achalasia, and produce a better clinical response during long-term follow-up compared with that in adults.


Outcomes of Gamma Knife Surgery retreatment for growing vestibular schwannoma and review of the literature.
Gamma Knife surgery has become an accepted treatment for small to medium‑size vestibular schwannoma with a high rate of tumor control and good clinical outcome. When GKS treatment fails to stop tumor growth, GKS retreatment can be proposed in selected cases. This retrospective study examines the clinical and tumor control outcome after the second GKS retreatment for the same vestibular schwannomas. A total of 14 consecutive vestibular schwannomas patients retreated with 2nd GKS were included: The median time interval between GKS treatments was 44 months, and the median follow‑up duration after last GKS retreatment was 60 months. The median marginal dose used for the first and second treatments was 12 Gy. The median tumor volume at the initial GKS was 2.4cc (range 0.27-3.8) and was 3.8cc (range 1.21-7.6) at the GKS retreatment. At the last follow‑up, 93% (13 patients) had tumor growth control, decreased in 4, remained unchanged in 9, and increased tumor size in one patient. New facial or severe trigeminal palsy did not occur after the second GKS retreatment. The hearing was not preserved except in one patient post-GKS retreatment. GKS retreatment after the failure of initial GKS to control vestibular schwannomas growth appears to be an effective strategy and can be proposed as an alternative to microsurgery when the tumor volume remains within the usual radiosurgical range.


MiR-375 promotes cisplatin sensitivity of lung adenocarcinoma.
Cisplatin-based chemotherapy has been widely used in the treatment of lung adenocarcinoma (LUAD). However, the development of cisplatin resistance becomes a major obstacle impeding the curative effect. It remains necessary to uncover the molecular mechanism of cisplatin resistance. Based on the CCLE database, lung cancer cell lines were divided into cisplatin-resistant and cisplatin-sensitive groups. The differentially expressed miRNAs were filtered and further identified by survival prognosis analysis. After transfection with miR-375 inhibitor or mimic, cell cytotoxicity assay, flow cytometry and western blot were conducted to validate the role of miR-375. The transcription factor (TF)-miRNA network was constructed based on TransmiR. The target genes of miR-375 were predicted by Starbase and further verified by RT-qPCR and immunohistochemistry results in the Human Protein Atlas. Functional enrichment analysis was performed with GO terms and KEGG. In this study, miR-375 showed the ability to promote cisplatin sensitivity and apoptosis of LUAD. Genes correlated with miR-375 in LUAD were analyzed and ABCC8 showed the strongest positive correlation. Moreover, transcription factors that regulate miR-375 expression were predicted. MBNL1, PTPN3, PRKD1 and RPN1 were identified as the target genes of miR-375. Enrichment analysis demonstrated that miR-375-related genes associated with promoting cell proliferation and anti-apoptosis were involved in the MAPK signaling pathway. Overall, this study provides new insights into the role of miR-375 in the cisplatin sensitivity of LUAD. Our present findings may serve as a theoretical basis for new therapeutic strategies and predictive models of cisplatin resistance in LUAD.


Determination of fexofenadine in human plasma and urine by liquid chromatography-mass spectrometry.
A sensitive method was developed to determine fexofenadine in human plasma and urine by HPLC-electrospray mass spectrometry with MDL 026042 as internal standard. Extraction was carried out on C18 solid-phase extraction cartridges. The mobile phases used for HPLC were: (A) 12 mM ammonium acetate in water and (B) acetonitrile. Chromatographic separation was achieved on a LUNA CN column (10 cm x 2.0 mm I.D., particle size 3 microm) using a linear gradient from 40% B to 60% B in 10 min. The mass spectrometer was operated in the selected ion monitoring mode using the respective MH+ ions, m/z 502.3 for fexofenadine and m/z 530.3 for the internal standard. The limit of quantification achieved with this method was 0.5 ng/ml in plasma and 1.0 ng in 50 microl of urine. The method described was successfully applied to the determination of fexofenadine in human plasma and urine in pharmacokinetic studies.


Biochemical characterization of interleukin 1 from a human monocytic cell line.
A protein with interleukin 1 (IL-1) activity was obtained from the human acute monocytic leukemia cell line (THP-1) and purified by ultrafiltration, Ultrogel AcA54 chromatography, isoelectric focusing, and discontinuous polyacrylamide gel electrophoresis. Like the pI 6.8 species of IL-1 from human peripheral blood monocytes (PBM), the cell line IL-1 has a molecular weight (MW) of 14,000, a pI of 6.8, is heat labile, and does not bind to concanavalin A-Sepharose. Chemical modification of arginine residues by phenylglyoxal or sulfhydryl groups by N-ethyl maleimide and iodoacetamide completely destroys the activity of IL-1 from THP-1 cells as well as that of the pI 6.8 component from PBM. In contrast, the pI 5.1 component of PBM IL-1 is resistant to heat denaturation and sulfhydryl reagents, although it is totally inactivated by phenylglyoxal. IL-1 from THP-1 cells enhanced the proliferative response of the same subpopulations of PNA- thymocytes as IL-1 from PBM. These observations suggest that IL-1 derived from this cell line is similar to the IL-1 pI 6.8 species produced by human monocytes, and distinct from the pI 5.1 species.


Hepatocyte nuclear factor 1 alpha influences pancreatic cancer growth and metastasis.
Hepatocyte nuclear factor 1 homeobox alpha (HNF1α) is a transcription factor involved in endodermal organogenesis and pancreatic precursor cell differentiation and development. Earlier studies have reported a role for HNF1α in pancreatic ductal adenocarcinoma (PDAC) but it is controversial. The mechanism by which it impacts PDAC is yet to be explored in depth. In this study, using the online databases we observed that HNF1α is upregulated in PDAC, which was also confirmed by our immunohistochemical analysis of PDAC tissue microarray. Silencing HNF1α reduced the proliferative, migratory, invasive and colony forming capabilities of pancreatic cancer cells. Key markers involved in these processes (pPI3K, pAKT, pERK, Bcl2, Zeb, Snail, Slug) were significantly changed in response to alterations in HNF1α expression. On the other hand, overexpression of HNF1α did not induce any significant change in the aggressiveness of pancreatic cancer cells. Our results demonstrate that reduced expression of HNF1α leads to inhibition of pancreatic cancer growth and progression, which indicates that it could be a potential oncogene and target for PDAC.


[Antibiotic sensitivity of important pathogens of bacterial respiratory tract infections in Northeast Germany].
There is still a lack of comprehensive study results about resistance of bacterial respiratory pathogens from the east of the Federal Republic of Germany. In Greifswald we isolated in 1995 and 1996 320 strains of typical pathogens and tested their susceptibility to 14 antibiotics, using the microbouillon dilution method. Pneumococci and beta-haemolytic streptococci were completely susceptible to penicillin and all other beta-lactam-antibiotics, clindamycin and vancomycin. 4.8% of pneumococci and 10.0% of Streptococcus pyogenes isolates were resistant to erythromycin. This should lead to a more differentiated use of macrolides as concerns these pathogens, especially as penicillin still has high efficacy in these cases. Pneumococci had resistance rates of 4.8%, 9.5% and 6.7% to cotrimoxazole, tetracycline and ofloxacin, respectively. No strain of Haemophilus influenzae produced a beta-lactamase, 2.6% of strains were relatively resistant to ampicillin, even in combination with sulbactam. Cefaclor, erythromycin and tetracycline had restricted efficacy of 16.7%, 14.1% and 53.8%, respectively. Cefuroxime, cefixime, cefepime, imipenem and ofloxacin all had complete efficacy. 77.8% of Moraxella catarrhalis isolates were beta-lactamase positive. Strains were susceptible to erythromycin, ofloxacin and all tested beta-lactam-antibiotics except ampicillin and cefaclor. There was a relative resistance to clindamycin and tetracyclin of 22.2% and 2.2%, respectively. We present our results in national and international comparison, describe tendencies of resistance, give reasons for the low incidence of penicillin resistance in pneumococci in Germany and draw conclusions for empirical chemotherapy.


All in the mind? Estimating the effect of mental health on health behaviours.
Health behaviours and mental illness have been found to be strongly correlated, both across and within individuals. However, establishing the precise nature of this link, particularly in terms of causality and its direction, remains a challenge. Much of the relevant existing research follow an experimental design and focus on measuring how changes in health behaviours affect mental health. The study examines changes in mental health and health behaviours within individuals across time and evaluates the evidence for a causal pathway from psychological distress to risky health behaviours. Using a population representative longitudinal survey providing data from Australian households, this study employs a fixed-effects panel model framework to account for unobserved time-invariant individual heterogeneity. An instrumental variable estimation strategy is employed to specifically evaluate the evidence of a causal pathway from psychological distress to risky health behaviours. The results confirm strong within-individual associations between psychological distress and health behaviours. Further evidence supports a causal relationship from psychological distress to health behaviours, though only for men and lifestyle habits. The case for a direct causal pathway is less straightforward when considering dietary habits, and potentially also for women. This lack of clarity suggests at least some of the observed within-individual associations reflect causal effects of psychological distress on risky health behaviours. Given the substantial burden of disease associated with unhealthy behaviours, further research examining potential causal pathways from mental health to health behaviours should be a priority, as there is the potential to reduce health burden through mental health improvements.


Use of Low-Molecular-Weight Heparin and Peak anti-Xa Monitoring In Severe SARS-CoV-2 Disease: A Brief Report.
In 2019, a novel coronavirus was identified in Wuhan, China. This strain was classified as a pandemic in early 2020 by the World Health Organization (WHO), rapidly reaching millions of cases worldwide and overwhelming intensive care units. One distinct feature identified in severe SARS-CoV-2 is abnormal and complex coagulation and hematologic abnormalities, including significantly elevated D-dimer and fibrin/fibrinogen values possibly increasing morbidity and mortality in this patient population. Aggressive anticoagulation therapy with appropriate peak anti-Xa level monitoring has produce satisfactory results at our institution. Our intent is to present a case series of our strategy to highlight the benefits of this approach.


Symptom dimensions of disruptive behavior disorders in adolescent drivers.
Adolescents with disruptive behavior disorder, including ADHD, are more likely to engage in risky driving practices and, consequently, are more likely to be involved in a motor vehicle crash (MVC) than their non-ADHD peers. It is unclear whether symptoms of inattention, hyperactivity/impulsivity, or oppositional defiant disorder (ODD) increase risk of poor driving outcomes. A total of 41 participants (16-19 years old) reported their ADHD and ODD symptoms and risky driving practices (errors and violations). History of citations and MVCs were acquired from state records. Relative predictive utility of symptom dimensions was assessed using multiple regressions. Inattention solely predicted driving variables of interest: Greater levels of inattention were predictive of more citations, MVCs, and self-reported errors and violations. Findings suggest that symptoms of inattention play a primary role in driving-related problems among adolescents. Implications for future research and practice are discussed.


T-wave morphology parameters based on principal component analysis reproducibility and dependence on T-offset position.
T-wave morphology parameters based on principal component analysis (PCA) are candidate to better understand the relation between QT prolongation and torsades de pointes. We aimed to assess the repeatability and to determine the influence of T-end position on PCA parameters. Digital ECGs recorded from 30 subjects were used to assess short term (5 minutes), circadian and long-term (28 days) repeatability of PCA parameters. The T-end cursor position was moved backward and forward (+/- 8 ms) from its optimal position. We calculated QRS-T angle, PCA ratio, and T-wave residuum (TWR). At long-term evaluation, coefficients of variation were 11.3 +/- 9.9%, 11.7 +/- 7.1%, and 23.0 +/- 22.0% for the QRS-T angle, PCA ratio, TWR, respectively. After moving the T-end cursor, repeatability was 0.42 +/- 0.2%, 1.00 +/- 1.04%, 4.0 +/- 4.2% for the same PCA parameters. T-wave morphology parameters based on PCA are reproducible with the exception of TWR and QRS-T angle. In addition, PCA is robust, showing only little dependence on T-end cursor position. These data should be taken into account for safety pharmacology trials.


A hand-ergonomics training kit: development and evaluation of a package to support improved awareness and critical thinking.
A need for a hand-ergonomics training kit has been identified to increase critical thinking concerning choice of hand tools. This study deals with the design, use and evaluation of a hand-ergonomics training kit for use in ergonomics training programmes. The effects on awareness of hand ergonomics among training course participants have been evaluated by means of a questionnaire and interviews at a car production plant in Sweden. The evaluation was carried out about one and a half years after training with the hand-ergonomics training kit. The training kit consists of a guide to practical exercises, equipment for measuring hand size and strength, examples of hand tools for use in practical exercises, equipment for testing and evaluating the hand tools and checklists and judgement forms for qualitative evaluation. In addition, the kit contains relevant scientifically based reference reports on hand ergonomics. The evaluation showed that the practical exercises with the hand-ergonomic training kit had, to a remarkable extent, increased individuals' awareness of anthropometric differences and of the importance of ergonomically well-designed hand tools. After the practical exercises with the training kit, communication within the plant when choosing hand tools seems to be based on objective criteria to a higher degree, however, the results indicate that this communication could be further improved.


Leucine-enkephalin- and neuropeptide Y-modulation of [3H]noradrenaline release in the oviduct of mature and juvenile rabbits.
1. The effects of leucine-enkephalin and neuropeptide Y (NPY) on [3H]noradrenaline release induced by electrical field stimulation were studied in the isthmic part of the oviduct of juvenile and mature rabbits. 2. [3H]noradrenaline and total tritium overflow in the presence of cocaine, corticosterone and hyoscine were determined by liquid scintillation spectrometry. 3. Tritium overflow evoked by electrical stimulation (1 or 4 Hz, 1 msec) was calcium dependent. [3H]noradrenaline content (measured by ion exchange chromatography) accounted for 85% of the total tritium overflow. 4. Leucine-enkephalin (1 microM) in the presence of the peptidase inhibitor bacitracin reduced the stimulation-evoked tritium overflow in mature rabbits by 26.1 +/- 1.6% and in juvenile rabbits by 11.9 +/- 1.9%. Naloxone (1 microM) antagonized the effect of leucine-enkephalin. 5. NPY (0.2 microM) reduced the evoked tritium overflow in mature rabbits by 23.4 +/- 2.4% and in juvenile rabbits by 17.2 +/- 4.3%. 6. It is concluded that leucine-enkephalin and NPY inhibited [3H]noradrenaline release in rabbit oviduct and the modulatory effect of leucine-enkephalin depends on maturity while NPY modulation is a more independent system.


Binding of fluoresceinated lectins to normal and dinitrofluorobenzene treated human leucocytes.
Using fluoresceinated lectins we have shown the receptor distribution on normal human granulocytes and lymphocytes following tagging with 1-fluoro- 2, 4-dinitrobenzene (DNFB). DNP-tagged cells exhibited strong, smooth membrane staining and produced smaller patches dispersed uniformly over the entire cell surface.


Effect of Hedan Tablets on Body Weight and Insulin Resistance in Patients with Metabolic Syndrome.
Apart from their recognized lipid-lowering effect, Hedan tablets, a mixture of Chinese herbal medicines, have demonstrated a certain weight-loss effect in clinical practice. The aim of this randomized, double-blind, placebo-controlled study was to verify the effect of Hedan tablets on body weight (BW) and insulin resistance (IR) in patients with metabolic syndrome (MetS). A total of 62 eligible patients with MetS were divided into two groups: the treatment group (Hedan tablets at 4.38 g/day tid) and the control group (placebo treatment). Both groups attended follow-ups at 8, 16, and 24 weeks during the process. The parameters of the assessment include lipid level, BW, triglyceride (TG) to high-density lipoprotein cholesterol (HDLc) ratio (TG/HDLc), homeostasis model assessment for IR (HOMA-IR) index, and adiponectin. Patients in the treatment group showed a significant decrease in BW compared to those in the control group (-4.47 vs. 0.06 kg) after 8 weeks of treatment. A significant decrease in body mass index (BMI) was also observed in the treatment group after 16 weeks of treatment (-1.79 vs. -0.03 kg/m2). In the treatment group, 20 out of 31 (64.5%) patients lost 5-10% BW and 4 out of 31 (12.9%) patients lost over 10% BW after 24 weeks of treatment. Although there were no significant changes in the patients' HOMA-IR, the treatment group showed a significant reduction in TG/HDLc (-0.98 vs. -0.19) after 8 weeks of treatment and a significant increase in adiponectin (6.87 vs. -0.43) after 16 weeks of treatment. The Hedan tablets significantly improve BW, BMI, TG/HDLc, and adiponectin in patients with MetS. Thus, Hedan tablets may be used as an adjunct to existing MetS management methods.


Corneal endothelial and intraocular pressure changes after phacoemulsification with Amvisc Plus and Viscoat.
A prospective, randomized, observer-marked study was conducted to compare corneal endothelial and intraocular pressure (IOP) changes after cataract surgery with the viscoelastic agents Amvisc Plus and Viscoat. Forty-nine patients (50 eyes) who had uncomplicated phacoemulsification and implantation of a posterior chamber intraocular lens were randomly assigned to either Amvisc Plus (25 eyes) or Viscoat (25 eyes) groups. Surgical technique was rigidly standardized in all cases. No significant difference in endothelial parameters or postoperative IOPs measured at 24 hours, one week, and two months was detected by analysis of variance between the Amvisc Plus and the Viscoat groups. Visualization was difficult with Viscoat because of its tendency to retain bubbles. Phacoemulsification energy was related to a loss of endothelial density, regardless of the viscoelastic used. The postoperative beta blocker may have contributed to the lower average postoperative IOP than has been reported.


Risk factors for the formation of a steinstrasse after extracorporeal shock wave lithotripsy: a statistical model.
We studied the various stone, renal and therapy factors that could affect steinstrasse formation after extracorporeal shock wave lithotripsy (ESWL), Dornier Medical Systems Inc., Marietta, Georgia to define the predictive factors for its formation. Thus, steinstrasse could be anticipated and prophylactically avoided. Between February 1989 and May 1999, 4,634 patients were treated with a Dornier MFL 5000 lithotriptor (Dornier Med Tech, GmbH, Germany). Renal stones were encountered in 3,403 patients and ureteral stones in 1,231. Steinstrasse were recorded in 184 patients, of whom 74 required intervention and formed the "complicated group." All patient data, stone and renal characteristics, and data of ESWL were reviewed. Univariate and multivariate statistical analyses of patients, stones and therapy characteristics in correlation with the incidence of steinstrasse formation were performed to assign the factors that had a significant impact on steinstrasse formation. The overall incidence of steinstrasse was 3.97%. The steinstrasse was in the pelvic ureter in 74% of the cases, lumbar ureter in 21.7% and iliac ureter in 4.3%. Steinstrasse incidence significantly correlated with stone size and site, the power level (kV.) used during therapy and radiological renal features. Steinstrasse was more common with renal stones more than 2 cm. in diameter in a dilated system, especially with the use of high power (greater than 22 kV.) for disintegration. A statistical model was constructed to estimate the risk of steinstrasse formation accurately. Stone size and site, renal morphology and shock wave energy are the significant predictive factors controlling steinstrasse formation. If a patient has a high probability of steinstrasse formation, close followup with early intervention or prophylactic pre-ESWL ureteral stenting is indicated.


Congenital cystic adenomatoid malformation - dangers of misdiagnosis: a case report.
Congenital cystic adenomatoid malformation is a rare pulmonary malformation, but is the most common lung malformation observed in children. In developing countries, such as Morocco, prenatal diagnosis is missing. Congenital cystic adenomatoid malformation may occur after birth in the presence of complications and needs a computed tomography scan for confirmation. However, our lack of awareness of this malformation has been responsible for a late and wrong diagnosis along with therapeutic errors. We report the first case in Morocco where diagnosis is confirmed by histology after death. A 10-month-old Arab boy was prescribed various antibiotics (including anti-staphylococcal) and endured repeated chest drainages, leading to his death just after radiological diagnosis and instant surgery. The goal of this case report is to firmly express the need for both pediatricians and radiologists to enlarge diagnosis investigations, especially of congenital or constitutional entities in children, as soon as recurrence of respiratory distress and pulmonary infections are manifested. We also emphasize this important entity because of its frequency, to avoid the eventual therapeutic errors.


Crohn's disease in the Hunter Valley region of Australia.
Between 1967 and 1991, 130 patients underwent surgery for Crohn's disease at The Royal Newcastle Hospital, New South Wales. There were more patients in recent years, which suggested an increased incidence of the condition (2.1/100 000/per year). The clinical profile and recurrence rate were similar to other studies. However, a greater proportion of patients required surgery for chronic small bowel obstruction and fulminant colitis compared with patients in other countries.


Orchestrated Strategies for Developing Fluorophores for NIR-II Imaging.
Fluorescence imaging (FLI), a non-invasive, real-time, and highly sensitive imaging modality, allows for investigating the molecular/cellular level activities to understand physiological functions and diseases. The emergence of the second near-infrared window (NIR-II, 1000-1700 nm) has endowed fluorescence imaging with deeper tissue penetration and unprecedented clarity. Among the various NIR-II imaging fluorophores, the organic fluorescent probes have occupied a pivotal position in bioimaging due to their higher biocompatibility, safety, and potential for clinical applications compared with those of the inorganic probes. To obtain high-quality organic dyes, diverse strategies have been taken. In this review, different strategies for optimizing NIR-II organic fluorophores are summarized, including traditional chemical modifications, and emerging bioengineering operations, which have not previously been elaborated on and summarized. Moreover, the bioengineering strategies are highlighted using endogenous serum proteins and even exogenous gene-editing proteins, which would provide fresh insights to design good-performance dyes and help develop NIR-II probes with clinical translation potential in the future. A critical perspective on the direction of the design strategies of NIR-II dyes for disease imaging is also proposed.


Mendelian randomization indicates that atopic dermatitis contributes to the occurrence of diabetes.
An association has been indicated between atopic dermatitis (AD), a prevalent chronic inflammatory skin disease, and diabetes mellitus. However, the exact causal relationship between AD and both type 1 diabetes (T1D) and type 2 diabetes (T2D) remains controversial. This study aimed to explore the causal association between AD and diabetes by Mendelian Randomization (MR) approaches. Public genetic summary data for AD was obtained from EAGLE study. Single nucleotide polymorphisms of diabetes were retrieved from four genome-wide association studies that had been performed in European populations. Inverse variance weighted (IVW) in MR analysis was used as the primary means of causality estimation. Several complementary analyses and sensitivity analyses were performed to calculate MR estimates and to enhance the causal inference, respectively. The R package 'TwoSampleMR' was used for analysis. Genetically predicted AD led to a higher risk of T1D (OR, 1.19; 95% CI, 1.05, 1.34; P = 0.006) and T2D (OR, 1.07; 95% CI, 1.02, 1.11; P = 0.003) based on random-effect IVW method. The complementary analyses provided similar positive results. Cochran's Q test and I2 statistics indicated moderate heterogeneity between AD and both T1D and T2D. No significant horizontal pleiotropy was detected by MR-Egger Intercept p except summary data from FinnGen consortium. Genetically predicted AD is a risk factor for both T1D and T2D. These findings imply potential shared pathological mechanisms between AD and diabetes, thus suggesting the significance of early clinical diagnosis and prevention of AD in reducing the incidence of diabetes.


Late bone metastasis in breast carcinoma.
A 49-year-old woman was free of metastases for 14 years after a left radical mastectomy. Metastases were found only in the skull, also the bone marrow biopsy demonstrated metastases.


Effects of high-fat diet and fasting on levels of acyl-coenzyme A binding protein in liver, kidney, and heart of rat.
Acyl-coenzyme A (CoA) binding protein (ACBP) is a 10-kd protein that binds acyl-CoA moieties and stimulates medium-chain fatty acid synthesis by goat mammary gland fatty acid synthetase. Its exact role in intermediary lipid metabolism has not been fully elucidated. It is hypothesized that ACBP is directly involved in the metabolism of lipid. In the present study, purified rat liver ACBP was used to generate a polyclonal antisera for radioimmunoassay of ACBP in tissue specimens isolated from fasted rats and rats fed normal rat chow and a high-fat diet. In addition, purified ACBP was used to examine its effect on the activity of mitochondrial outer membrane (OM) carnitine palmitoyltransferase (CPT0). Fasting for 24 hours significantly decreased tissue levels of ACBP in the liver (69.0 +/- 7.2 v 46.7 +/- 5.0 pg/ng DNA), whereas feeding of a high-fat diet for 48 hours caused ACBP levels to increase (69.0 +/- 7.2 v 103.9 +/- 18.0). Hepatic levels of this protein continued to increase and remained elevated with prolonged exposure to the high-fat diet (28 days). A similar pattern of change was observed in the kidney, but the magnitude of change was less. Heart ACBP did not respond acutely to the high-fat diet, but did increase after prolonged exposure (28 days). Fasting had no effect on ACBP levels in kidney and heart. Addition of ACBP to an in vitro assay system significantly increased the activity of CPT0 (from 5.2 +/- 0.8 to 72.1 +/- 5.3 nmol palmitoylcarnitine formed.min-1.mg-1 protein) when measured under inhibiting concentrations of palmitoyl-CoA (40 mumol/L).(ABSTRACT TRUNCATED AT 250 WORDS)


Recto-femoral fistula presenting as emphysematous cellulitis of the knee: a case report & literature review.
Purpose. The rectofemoral fistula represents a devastating complication of colorectal surgery. Its early diagnosis and treatment are critical to obtain a good patient outcome. Case Presentation. A 75-year-old Caucasian female patient presented with high fever, ileus, low back pain, sciatic nerve palsy, and infection of the right knee. After numerous surgical debridements and antibiotic therapies, a rectofemoral fistula was diagnosed. Conclusion. Increased doctors' alertness is mandatory for the early identification and surgical treatment of patients suffering from a rectofemoral fistula before the stage of diffuse infection has significantly decreases their postoperative morbidity and mortality.


Normal rat intestinal cells IEC-18: characterization and transfection with immortalizing oncogenes.
IEC-18 cells, a cell line derived from the ileum of rat intestine, have the characteristics of normal cells since they have a contact inhibited cell growth, do not form colonies in soft agar and are not tumorigenic when injected in nude mice. IEC-18 cells were transfected with nuclear oncogenes, c-myc, v-myc and SV40 T antigen in order to obtain immortal cell lines. Independent clones were isolated and characterized for the growth properties. Expression of v-myc altered the morphology of the cells and shortened the doubling time. A slow growth together with a low cloning efficiency was associated with the expression of SV40 T antigen. No changes either in growth or in morphology were observed in c-myc-expressing IEC-18 cells. Expression of these nuclear oncogenes did not result in the neoplastic transformation of the IEC-18 cells, since none of the clones lost the anchorage dependence or were able to form tumors in vivo. The c-myc-containing IEC-18 cells were unable to secrete in the growth medium TGF α and exposure to TGF β inhibited the growth rate by 30%. All these observations are consistent with the conclusion that the expression of nuclear oncogenes does not lead to the neoplastic transformation of these cells.


Genetic diversity of Tn1546-like elements in clinical isolates of vancomycin-resistant enterococci.
We have investigated the genetic diversity of Tn1546 among 17 vancomycin-resistant enterococci (VRE) isolates of Enterococcus faecium. Most of these multidrug-resistant strains harboured plasmids of 2 kb to >300 kb in size. The vancomycin resistance marker vanA was located on both the plasmid and the chromosomal DNA. VRE isolates 18 and 22 failed to amplify the orf1-IR(R) and orf2-IR(R) but contained the orf1 and orf2. VRE3 failed to amplify the orf1, orf2, vanR and vanS, but still yielded a larger than expected (4.4 kb vs. 2.3 kb) vanSH amplicon. VRE9, 10, 21 and 22 also yielded larger (5.5 kb) vanSH amplicons; all others yielded 4.0 kb vanSH amplicons. Sequence analysis of the vanSH amplicons from VRE9, 10, 21 and 22 revealed the presence of IS1251 between the vanS and vanH genes in these isolates. The observed vanSH amplicon from VRE3 contained orf31, orf30 and orf29 of the plasmid pRUM followed by the vanHAXYZ region of Tn1546. Translocation of Tn1546 to a pRUM-like plasmid in VRE3 resulted in the loss of its orf1, orf2, vanR and vanS elements and a loss of the orf32 of pRUM, leading to a unique structural arrangement of vanA elements that is hitherto unknown.


A novel adaptive neural network-based time-delayed estimation control for nonlinear systems subject to disturbances and unknown dynamics.
This paper presents an adaptive backstepping-based model-free control (BSMFC) for general high-order nonlinear systems (HNSs) subject to disturbances and unstructured uncertainties to enhance the system tracking performance. The proposed methodology is constructed based on the backstepping control (BSC) with radial basis function neural network (RBFNN) -based time-delayed estimation (TDE) to overcome the obstacle of unknown system dynamics. Additionally, a command-filtered (CF) approach is involved to address the complexity explosion of the BSC design. As the errors arising from approximation, new control laws are established to reduce the effects in this regard. The stability of the closed-loop system is guaranteed through the Lyapunov theorem and the superiority of the proposed methodology is confirmed through a comparative simulation with other model-free approaches.


Development, hatching, and infectivity of Echinostoma caproni (Trematoda) eggs, and histologic and histochemical observations on the miracidia.
Light microscopy studies were done on the eggs and miracidia of the intestinal trematode Echinostoma caproni. Eggs were obtained by homogenizing adult worms derived from ICR mice in artificial spring water (ASW). To study egg development and to obtain miracidia, eggs were incubated in ASW in either a light or a dark environment for 10 days at 28 degrees C. Egg development was characterized and staged at 2-day intervals from day 0 to day 10 postembryonation. No significant difference in development was seen in eggs maintained under conditions of light or darkness. Eggs maintained in darkness for 10 days and exposed to incandescent light produced a large synchronous hatch of miracidia within 3 h of exposure to light. As expected, miracidia used to expose Biomphalaria glabrata snails produced patent infections at 5-7 weeks postinfection (p.i.). Embryonated eggs aged from 0 to 10 days that had been fed to B. glabrata snails also produced infections within 4 weeks of infection. Miracidial longevity studies showed that 50 newly hatched miracidia maintained at either 4 degrees, 12 degrees, 22 degrees, or 38 degrees C lived for 6, 28, 14, and 5 h, respectively. Histology studies were done on whole miracidia stained in 0.01% neutral red and in Schneider's acetocarmine. Histochemistry studies of miracidia stained with alcian blue (pH 2.5) showed the presence of acid mucopolysaccharides in the epidermal plates; miracidia treated with periodic acid-Schiff plus 0.5% malt diastase showed the presence of glycogen in the body and in the epidermal/subepidermal region. Miracidia stained with Lillie's Oil Red O showed the presence of sparse neutral fat droplets in the body.


Graphene oxide/chitosan composites as novel support to provide high yield and stable formulations of pectinase for industrial applications.
An extracellular pectinase from a mixed consortium of Bacillus sp. (BSP) was immobilized onto graphene oxide/chitosan composite (GO/CS) through covalent binding to enhance its recycling and operational stability features. Different parameters were optimized, including cross-linker concentration (%), time, pH, and GO/CS-pectinase ratios. GO/CS-pectinase was further characterized by FT-IR and XRD. The activity of GO/CS-pectinase was reached up to 804 μmolmin-1 with an immobilization efficiency of 80.64 ± 1.15 % under optimum conditions. GO/CS-pectinase exhibited a 3.0-folds higher half-life (t1/2) than free pectinase at 50, 55, and 60 °C, respectively. The Vmax and KM values of GO/CS-pectinase were found to be nearly equal to the free pectinase indicating that conformational flexibility was retained. Kd, t1/2, ∆G*, ∆H*, and ∆S* of both free pectinase and GO/CS-pectinase was 0.0339 & 0.0721 min-1, 9.62 and 40.44 min, 81.35, 90.72 kJmol-1, 47.098 & 63.635 kJmol-1, -102.86 & -81.340 Jmole-1 K-1. SEM morphological analysis further confirmed the successful binding of pectinase with GO/CS, which retained about 92 % of its original catalytic activity after ten consecutive reaction cycles. Finally, GO/CS-pectinase was employed for guava juice clarification which exhibited the turbidity reduction up to 81 % after 75 min of treatment.


Increased airway responsiveness, allergy-type-I skin responses and systemic anaphylaxis in a humanized-severe combined immuno-deficiency mouse model.
In patients with allergic bronchial asthma, a strong relationship between elevated serum IgE antibody titres and the development of increased airway responsiveness (AR) has been demonstrated. To further elucidate the relationship between human (hu) IgE and development of increased AR, we developed an in vivo model utilizing immuno-compromised severe combined immuno-deficiency (SCID) mice. SCID mice were either reconstituted with peripheral blood mononuclear cells (PBMC) from non-atopic, healthy or atopic individuals sensitized against house dust mite allergen (Der p), or passively sensitized with plasma from non-atopic, healthy or atopic individuals. In both systems, atopic hu-SCID mice developed increased AR. The following results suggest that these responses were mediated via IgE antibodies: increased AR did not occur after transfer of either PBMC or IgE-negative plasma from non-atopic individuals; increased AR occurred simultaneous with increased serotonin release detected 15 min after allergen-aerosol challenge in bronchoalveolar lavage fluid; and increased AR required at least two allergen-aerosol challenges. SCID mice reconstituted with serum containing anti-Der p IgE antibodies developed positive immediate-type skin test responses to intradermal injection of Der p as well as anti-hu-IgE antibody. In addition, IgE binding to skin mast cells was demonstrated by immunohistochemistry. Furthermore, intravenous challenge of hu anti-Der p positive SCID mice with Der p resulted in systemic anaphylaxis. These data provide evidence that passive immunization of SCID mice with hu IgE alters AR and that T cells and eosinophils were not a requirement for the development of increased AR in this model.


The Debate on the Moral Responsibilities of Online Service Providers.
Online service providers (OSPs)-such as AOL, Facebook, Google, Microsoft, and Twitter-significantly shape the informational environment (infosphere) and influence users' experiences and interactions within it. There is a general agreement on the centrality of OSPs in information societies, but little consensus about what principles should shape their moral responsibilities and practices. In this article, we analyse the main contributions to the debate on the moral responsibilities of OSPs. By endorsing the method of the levels of abstract (LoAs), we first analyse the moral responsibilities of OSPs in the web (LoAIN). These concern the management of online information, which includes information filtering, Internet censorship, the circulation of harmful content, and the implementation and fostering of human rights (including privacy). We then consider the moral responsibilities ascribed to OSPs on the web (LoAON) and focus on the existing legal regulation of access to users' data. The overall analysis provides an overview of the current state of the debate and highlights two main results. First, topics related to OSPs' public role-especially their gatekeeping function, their corporate social responsibilities, and their role in implementing and fostering human rights-have acquired increasing relevance in the specialised literature. Second, there is a lack of an ethical framework that can (a) define OSPs' responsibilities, and (b) provide the fundamental sharable principles necessary to guide OSPs' conduct within the multicultural and international context in which they operate. This article contributes to the ethical framework necessary to deal with (a) and (b) by endorsing a LoA enabling the definition of the responsibilities of OSPs with respect to the well-being of the infosphere and of the entities inhabiting it (LoAFor).


FGF-2 modulates expression and distribution of GAP-43 in frog retinal ganglion cells after optic nerve injury.
Basic fibroblast growth factor (bFGF or FGF-2) has been implicated as a trophic factor that promotes survival and neurite outgrowth of neurons. We found previously that application of FGF-2 to the proximal stump of the injured axon increases retinal ganglion cell (RGC) survival. We determine here the effect of FGF-2 on expression of the axonal growth-associated phosphoprotein (GAP)-43 in retinal ganglion cells and tectum of Rana pipiens during regeneration of the optic nerve. In control retinas, GAP-43 protein was found in the optic fiber layer and in optic nerve; mRNA levels were low. After axotomy, mRNA levels increased sevenfold and GAP-43 protein was significantly increased. GAP-43 was localized in retinal axons and in a subset of RGC cell bodies and dendrites. This upregulation of GAP-43 was sustained through the period in which retinal axons reconnect with their target in the tectum. FGF-2 application to the injured nerve, but not to the eyeball, increased GAP-43 mRNA in the retina but decreased GAP-43 protein levels and decreased the number of immunopositive cell bodies. In the tectum, no treatment affected GAP-43 mRNA but FGF-2 application to the axotomized optic nerve increased GAP-43 protein in regenerating retinal projections. We conclude that FGF-2 upregulates the synthesis and alters the distribution of the axonal growth-promoting protein GAP-43, suggesting that it may enhance axonal regrowth.


Nitric oxide: a regulator of mucosal defense and injury.
There is an abundance of evidence that nitric oxide plays a critical role in regulating several components of gastrointestinal mucosal defense. Suppression of nitric oxide synthesis increases susceptibility to injury, while administration of nitric oxide donors increases resistance to injury. On the other hand, nitric oxide has been implicated as a mediator of tissue injury in the gastrointestinal tract during inflammatory reactions. In these cases, the nitric oxide is generally believed to be derived from an inducible isoform of nitric oxide synthase. In this review, we provide an overview of the evidence for and against these dual roles of nitric oxide in modulating gastrointestinal mucosal defense and injury. We also highlight the potential therapeutic benefits that may be realized through modulation of tissue nitric oxide levels.