The Role of Ataxia Telangiectasia Mutant and Rad3-Related DNA Damage Response in Pathogenesis of Human Papillomavirus.
Human papillomavirus (HPV) infection leads to a variety of benign lesions and malignant tumors such as cervical cancer and head and neck squamous cell carcinoma. Several HPV vaccines have been developed that can help to prevent cervical carcinoma, but these vaccines are only effective in individuals with no prior HPV infection. Thus, it is still important to understand the HPV life cycle and in particular the association of HPV with human pathogenesis. HPV production requires activation of the DNA damage response (DDR), which is a complex signaling network composed of multiple sensors, mediators, transducers, and effectors that safeguard cellular DNAs to maintain the host genome integrity. In this review, we focus on the roles of the ataxia telangiectasia mutant and Rad3-related (ATR) DNA damage response in HPV DNA replication. HPV can induce ATR expression and activate the ATR pathway. Inhibition of the ATR pathway results in suppression of HPV genome maintenance and amplification. The mechanisms underlying this could be through various molecular pathways such as checkpoint signaling and transcriptional regulation. In light of these findings, other downstream mechanisms of the ATR pathway need to be further investigated for better understanding HPV pathogenesis and developing novel ATR DDR-related inhibitors against HPV infection.


Surgical technique of placement of an external jugular tunneled hemodialysis catheter in a large pig model.
Currently, there is insufficient knowledge about the surgical anatomy and surgical techniques in large animals that can be used to test medical devices designed for human use. We encountered this problem in our study requiring the placement of jugular vein, tunneled, cuffed hemodialysis catheter in 70 kg pigs. Despite the operator's extensive expertise in placing tunneled hemodialysis catheters in humans, the important differences in anatomy made the procedure and choosing the appropriate catheter length challenging. The following article describes the anatomy and our technique for the placement of tunneled hemodialysis catheter in the pig model. We consider our surgical technique to be sound because in all animals the catheters were placed in the desired location, the procedures were well tolerated by the animals, and there were no immediate or late complications. We present our experience to help other researchers who might encounter the same problem.


Maize starch derived boron doped carbon spheres via facile solvothermal route as the photoluminescence sensor for determination of pH and Cr(VI).
In this work, a kind of boron doped carbon spheres (B-CSs) was successfully synthesized utilizing maize starch as carbon source and boric acid as dopant via facile solvothermal method. The chemical structure of the prepared B-CSs was systemically investigated by TEM, FT-IR, XRD, XPS and EDS. The synthesized B-CSs feature spherical structure with average size of ∼254 nm and exhibit strong photoluminescence (PL) with maximum emission at a wavelength of ∼453 nm under irradiation at 350 nm, leading to a quantum yield of 6.2%. Furthermore, the aqueous pH and Cr(VI) has a significantly various impact on the PL intensity of B-CSs, which can be flexibly utilized as the PL sensor for detection aqueous pH and Cr(VI) in aqueous. Particularly, the B-CSs have a desirable sensitivity and selectivity for detection of Cr(VI) with a low detection limit of ∼0.34μmol l-1. Conclusively, our work provides a novel and dual-functional fluorescent sensor for detection of the pH and toxic metal ions in water environment.


Drug conjugates such as Antibody Drug Conjugates (ADCs), immunotoxins and immunoliposomes challenge daily clinical practice.
Drug conjugates have been studied extensively in preclinical in vitro and in vivo models but to date only a few compounds have progressed to the clinical setting. This situation is now changing with the publication of studies demonstrating a significant impact on clinical practice and highlighting the potential of this new class of targeted therapies. This review summarizes the pharmacological and molecular background of the main drug conjugation systems, namely antibody drug conjugates (ADCs), immunotoxins and immunoliposomes. All these compounds combine the specific targeting moiety of an antibody or similar construct with the efficacy of a toxic drug. The aim of this strategy is to target tumor cells specifically while sparing normal tissue, thus resulting in high efficacy and low toxicity. Recently, several strategies have been investigated in phase I clinical trials and some have entered phase III clinical development. This review provides a detailed overview of various strategies and critically discusses the most relevant achievements. Examples of the most advanced compounds include T-DM1 and brentuximab vedotin. However, additional promising strategies such as immunotoxins and immunoliposmes are already in clinical development. In summary, targeted drug delivery by drug conjugates is a new emerging class of anti-cancer therapy that may play a major role in the future.


Changes in lipid and lipoprotein levels after weight training.
Lipid and lipoprotein levels were prospectively studied in previously sedentary men (mean age, 33 years) and women (mean age, 27 years) undergoing 16 weeks of weight-training exercise. Women demonstrated a 9.5% reduction of cholesterol, 17.9% decrease in low-density lipoprotein (LDL) cholesterol and 28.3% lowering of triglycerides. The ratios of total cholesterol-high-density lipoprotein (HDL) cholesterol and LDL cholesterol-HDL cholesterol were reduced 14.3% and 20.3%, respectively. Among men, LDL cholesterol was reduced 16.2%, while the ratios of total cholesterol-HDL cholesterol and LDL cholesterol-HDL cholesterol were lowered 21.6% and 28.9%, respectively. Weight-training exercise appears to result in favorable changes in lipid and lipoprotein levels in previously sedentary men and women.


Eye Pain and Dry Eye in Patients with Fibromyalgia.
The aim of this study was to examine visual function and eye symptoms in fibromyalgia patients, with a particular focus on dry eye syndrome and eye pain. A tertiary care center-based cross-sectional study was carried out in chronic musculoskeletal pain patients diagnosed with fibromyalgia. Chronic musculoskeletal pain patients without fibromyalgia were enrolled as a comparison group. Self-reported eye pain was investigated with the McGill pain questionnaire and the numeric rating scale. In addition, we assessed corrected visual acuity, vision-related quality of life, and self-reported dry eye syndrome. A total of 90 musculoskeletal pain patients were included, with 66 patients fulfilling American College of Rheumatology 1990 criteria for fibromyalgia. Sixty-seven percent (95% confidence interval [CI] = 56%-78%) of the fibromyalgia patients reported eye pain, and 62% (95% CI = 43%-81%) of those were without fibromyalgia diagnosis. Sixty-seven percent (95% CI = 56%-78%) of the fibromyalgia patients reported an experience of dry eye compared with 76% (95% CI = 57%-95%) in the nonfibromyalgia group. Vision-related quality of life was noticeably reduced in both groups. Eye pain and dry eye are common in chronic pain patients, with comparable prevalence in musculoskeletal pain patients with and without fibromyalgia.


Contrast enhancement of cerebrospinal fluid on delayed MRI in a patient with an epidural abscess and renal failure.
Cerebrospinal fluid (CSF) is normally of low signal intensity on T1 weighted MRI. The case of a patient with renal failure who developed an epidural abscess, and in whom the CSF appeared of high signal intensity on delayed T1 weighted MRI is presented. Possible mechanisms for this are discussed.


Veno-arterial extracorporeal membrane oxygenation for drug intoxications: A single center, 14-year experience.
Acute cardiovascular failure remains a leading cause of death in severe poisonings. Veno-arterial extracorporeal membrane oxygenation (VA-ECMO) has been increasingly used as a rescue therapeutic option for those cases refractory to optimal conventional treatment. We sought to evaluate the outcomes after VA-ECMO used for drug intoxications in a single-center experience. We performed an observational analysis of our prospective institutional database. The primary endpoint was survival to hospital discharge. Between January 2007 and December 2020, 32 patients (mean age: 45.4 ± 15.8 years; 62.5% female) received VA-ECMO for drug intoxication-induced refractory cardiogenic shock (n = 25) or cardiac arrest (n = 7). Seven (21.8%) patients developed lower limb ischemia during VA-ECMO support. Twenty-six (81.2%) patients were successfully weaned after a mean VA-ECMO support of 2.9 ± 1.3 days. One (3.1%) patient died after VA-ECMO weaning for multiorgan failure and survival to hospital discharge was 78.1% (n = 25). In-hospital survivors were discharged from hospital with a good neurological status. Survival to hospital discharge was not statistically different according to sex (male = 75.0% vs. female = 80.0%; p = .535), type of intoxication (single drug = 81.8% vs. multiple drugs = 76.1%; p = .544) and location of VA-ECMO implantation (within our center = 75% vs. peripheral hospital using our Mobile Unit of Mechanical Circulatory Support = 100%; p = .352). Survival to hospital discharge was significantly lower in patients receiving VA-ECMO during on-going cardiopulmonary resuscitation (42.8% vs. 88.0%; p = .026). VA-ECMO appears to be a feasible therapeutic option with a satisfactory survival rate and acceptable complications rate in poisonings complicated by refractory cardiogenic shock or cardiac arrest.


First-order phase transition of fixed connectivity surfaces.
We report numerical evidence of the discontinuous transition of a tethered membrane model which is defined within a framework of the membrane elasticity of Helfrich. Two kinds of phantom tethered membrane models are studied via the canonical Monte Carlo simulation on triangulated fixed connectivity surfaces of spherical topology. One surface model is defined by the Gaussian term and the bending energy term, and the other, which is tensionless, is defined by the bending energy term and a hard wall potential. The bending energy is defined by using the normal vector at each vertex. Both models undergo the first-order phase transition characterized by a gap of the bending energy. The phase structure of the models depends on the choice of discrete bending energy.


Chemical genetics strategy to profile kinase target engagement reveals role of FES in neutrophil phagocytosis.
Chemical tools to monitor drug-target engagement of endogenously expressed protein kinases are highly desirable for preclinical target validation in drug discovery. Here, we describe a chemical genetics strategy to selectively study target engagement of endogenous kinases. By substituting a serine residue into cysteine at the DFG-1 position in the ATP-binding pocket, we sensitize the non-receptor tyrosine kinase FES towards covalent labeling by a complementary fluorescent chemical probe. This mutation is introduced in the endogenous FES gene of HL-60 cells using CRISPR/Cas9 gene editing. Leveraging the temporal and acute control offered by our strategy, we show that FES activity is dispensable for differentiation of HL-60 cells towards macrophages. Instead, FES plays a key role in neutrophil phagocytosis via SYK kinase activation. This chemical genetics strategy holds promise as a target validation method for kinases.


Grandisines C-G, indolizidine alkaloids from the Australian rainforest tree Elaeocarpus grandis.
Five new indolizidine alkaloids, grandisines C, D, E, F, and G (4-8), and one known indolizidine alkaloid, (-)-isoelaeocarpiline (3), were isolated from the leaves of Elaeocarpus grandis and their structures determined by 1D and 2D NMR spectroscopy. Grandisine C (4) is isomeric with the known compound rudrakine (1). The absolute configuration of grandisine D (5) was deduced by its conversion to (-)-isoelaeocarpiline. Grandisine E (6) contains a novel tetracyclic ring system. Grandisine F (7) is the 14-amino analogue of grandisine C. Grandisine G (8) contains the novel combination of a piperidine attached to an indolizidine. Grandisines C, D, F, and G and (-)-isoelaeocarpiline showed receptor binding affinity for the human delta-opioid receptor with IC(50) values of 14.6, 1.65, 1.55, 75.4, and 9.9 microM, respectively.


Best practices for working with rape crisis centers to address elder sexual abuse.
Sexual violence perpetrated on elders is a significant problem. Elder sexual assault victims often need more assistance than younger victims, yet they typically receive fewer services. Rape crisis centers have faced a multitude of barriers in addressing elder sexual abuse. This article provides an overview of the rape crisis movement and discusses why elders have been both overlooked as potential targets of sexual assault and underserved as victims. Recommendations are made to encourage collaboration between elder and rape crisis advocates. This article describes a Pennsylvania project to cross-train Department of Aging protective service workers and rape crisis advocates.


A Chemist's Perspective on the Role of Phosphorus at the Origins of Life.
The central role that phosphates play in biological systems, suggests they also played an important role in the emergence of life on Earth. In recent years, numerous important advances have been made towards understanding the influence that phosphates may have had on prebiotic chemistry, and here, we highlight two important aspects of prebiotic phosphate chemistry. Firstly, we discuss prebiotic phosphorylation reactions; we specifically contrast aqueous electrophilic phosphorylation, and aqueous nucleophilic phosphorylation strategies, with dry-state phosphorylations that are mediated by dissociative phosphoryl-transfer. Secondly, we discuss the non-structural roles that phosphates can play in prebiotic chemistry. Here, we focus on the mechanisms by which phosphate has guided prebiotic reactivity through catalysis or buffering effects, to facilitating selective transformations in neutral water. Several prebiotic routes towards the synthesis of nucleotides, amino acids, and core metabolites, that have been facilitated or controlled by phosphate acting as a general acid-base catalyst, pH buffer, or a chemical buffer, are outlined. These facile and subtle mechanisms for incorporation and exploitation of phosphates to orchestrate selective, robust prebiotic chemistry, coupled with the central and universally conserved roles of phosphates in biochemistry, provide an increasingly clear message that understanding phosphate chemistry will be a key element in elucidating the origins of life on Earth.


[Mandibular localization of thesaurismosis (polyvinyl-pyrrolidine) (author's transl)].
The authors report on a case of isolated mandibular localization of polyvinyl-pyrrolidine thesaurismosis. The patient, a man of 38, had been treated with injections of posterior pituitary extract, for a period of eighteen months, 10 year ago. Localization in the mandible is extremely rare, making diagnosis difficult unless a detailed anamnesis is available.


Noninvasive intrasac pressure measurement and the influence of type 2 and type 3 endoleaks in an animal model of abdominal aortic aneurysm.
The objective of this study was to noninvasively detect pressure changes within an excluded aneurysm sac in an animal model of abdominal aortic aneurysm (AAA) and to study the influence of type 2 and 3 endoleaks. A porcine model of AAA that allows for the creation of type 2 and 3 endoleaks was used. A miniaturized pressure monitoring device (3 x 9 x 1.5 mm; Remon Medical Technologies, Caesarea, Israel) was implanted within the surgically created and excluded aneurysm sac. The pressure monitoring device is an ultrasound-based system that allows for pressure measurements in a noninvasive, transcutaneous fashion. In addition, catheter-based pressures were taken within the aorta and directly in the AAA sac. Noninvasive measurements were taken in a transcutaneous fashion between the initial operation and the time of sacrifice, when the type 3 endoleak was created (2 weeks). The median mean arterial pressure was 66 mm Hg (range 55-120 mm Hg; N = 8). The median noninvasive sac pressure with a type 2 endoleak was 48 mm Hg (range 39-90 mm Hg; N = 8) and was almost identical to the catheter-based measurements. Noninvasive pressures could be measured as early as postprocedure day 1. Two animals had follow-up that suggested closure of the type 2 endoleak during the observation period. With the creation of the type 3 endoleak, the catheter and noninvasive sac pressure and waveform changed from a flatline trace to a higher-pressure pulsatile trace (median 54 mm Hg; range 46-81 mm Hg; N = 8), reproducing the arterial pressure and waveform. This is the first study, to our knowledge, that demonstrates the efficacy of a noninvasive, miniaturized pressure monitoring device in identifying pressure changes in an excluded aneurysm sac with type 2 and type 3 endoleaks. This technology holds great promise for follow-up of patients and identification of sac pressure changes after EVAR and may allow a change in the current follow-up strategy.


Ancestry-driven metabolite variation provides insights into disease states in admixed populations.
Metabolic pathways are related to physiological functions and disease states and are influenced by genetic variation and environmental factors. Hispanics/Latino individuals have ancestry-derived genomic regions (local ancestry) from their recent admixture that have been less characterized for associations with metabolite abundance and disease risk. We performed admixture mapping of 640 circulating metabolites in 3887 Hispanic/Latino individuals from the Hispanic Community Health Study/Study of Latinos (HCHS/SOL). Metabolites were quantified in fasting serum through non-targeted mass spectrometry (MS) analysis using ultra-performance liquid chromatography-MS/MS. Replication was performed in 1856 nonoverlapping HCHS/SOL participants with metabolomic data. By leveraging local ancestry, this study identified significant ancestry-enriched associations for 78 circulating metabolites at 484 independent regions, including 116 novel metabolite-genomic region associations that replicated in an independent sample. Among the main findings, we identified Native American enriched genomic regions at chromosomes 11 and 15, mapping to FADS1/FADS2 and LIPC, respectively, associated with reduced long-chain polyunsaturated fatty acid metabolites implicated in metabolic and inflammatory pathways. An African-derived genomic region at chromosome 2 was associated with N-acetylated amino acid metabolites. This region, mapped to ALMS1, is associated with chronic kidney disease, a disease that disproportionately burdens individuals of African descent. Our findings provide important insights into differences in metabolite quantities related to ancestry in admixed populations including metabolites related to regulation of lipid polyunsaturated fatty acids and N-acetylated amino acids, which may have implications for common diseases in populations.


Glial cell line-derived neurotrophic factor-mediated enteric neuronal survival involves glycogen synthase kinase-3beta phosphorylation and coupling with 14-3-3.
Glial cell line-derived neurotrophic factor (GDNF) promotes the growth and survival of enteric neurons, but the mechanisms involved are poorly understood. GDNF is known to promote the survival of enteric neurons through activation of the PI3-Kinase/Akt signaling pathway. We investigated the role of glycogen synthase kinase-3beta (GSK-3beta) in enteric neuronal survival, and the ability of GDNF to regulate the activity of GSK-3beta using primary rat embryonic enteric neurons. GDNF, through activation of the PI3-kinase pathway enhanced the phosphorylation of GSK-3beta at its N-terminal serine-9 residue, and promoted the association of GSK-3beta with 14-3-3. Transfection of a constitutively active S9A-GSK-3beta mutant prevented the survival effects of GDNF, whereas a dominant negative GSK-3beta construct prevented GDNF withdrawal-induced cell death. Increased GSK-3beta activity was associated with an increase in tau phosphorylation. Thus, GDNF promotes enteric neuronal survival by modulating GSK-3beta and its downstream target tau. Inhibitors of GSK-3beta activity may have therapeutic potential in improving enteric neuronal survival.


New Horizons: COVID-19 and the Burden of Neuropsychiatric Illness in Pakistan.
This manuscript reviews the current state of knowledge about the burden of mental illness and assesses the impact of COVID-19 illness on mental health in Pakistan. For this we analyzed secondary data obtained from the Institute of Health Metrics and Evaluation. The Global Burden of Disease (GBD) study draws from a wide range of data sources to quantify global and regional effects of a disease. We also did a literature search on the effects of COVID-19 illness on mental health and the psychosocial effects of COVID-19 and other Corona virus related illnesses such as SARS-CoV and MERS-CoV. Data from the studies obtained was utilized to extrapolate the anticipated effects of COVID-19 illness on healthcare workers, COVID-19 patients and the general public in Pakistan. Mental illness poses a significant challenge to Pakistan's under resourced health care system. COVID-19 has the potential to strain Pakistan's healthcare system to the breaking point. So far, the general morbidity from COVID-19 illness in Pakistan has been low compared to other countries but this could change in the coming weeks and months. Hidden within this crisis are also some opportunities for both healthcare and education.


Effect of intra-articular administration of buprenorphine after arthrocentesis of the temporomandibular joint: a pilot study.
The purpose of this double-blind study was to evaluate the effect of intra-articular opioid injections on postoperative pain and joint mobility after arthrocentesis of the temporomandibular joint (TMJ). Forty patients suffering from persistent pain in combination with anterior disc displacement (22 with reduction, 18 without reduction) and unresponsive to nonsurgical therapy underwent arthrocentesis of the affected TMJ under local anesthesia. All patients were randomly assigned to 2 groups. In group 1 (20 patients), 1 mL of buprenorphine solution was injected into the joint space at the end of the intervention; in group 2 (20 patients), 1 mL of saline was used. For postoperative pain management, all patients received Paracetamol ad libitum. Preoperatively and at postoperative days 2, 4, 7, and 14, individual pain levels were determined on a visual analogue scale together with maximum mouth opening. At days 2 and 4 after the intervention, remarkable differences were detectable between the 2 groups, showing a rapid normalization of joint mobility and pain scores in group 1; in contrast, improvement in group 2 was delayed. Intra-articular injections of long lasting opioids seem to influence joint mobility and pain positively within the first week after arthrocentesis.


Bioavailability of silver nanoparticles and ions: from a chemical and biochemical perspective.
Owing to their antimicrobial properties, silver nanoparticles (NPs) are the most commonly used engineered nanomaterial for use in a wide array of consumer and medical applications. Many discussions are currently ongoing as to whether or not exposure of silver NPs to the ecosystem (i.e. plants and animals) may be conceived as harmful or not. Metallic silver, if released into the environment, can undergo chemical and biochemical conversion which strongly influence its availability towards any biological system. During this process, in the presence of moisture, silver can be oxidized resulting in the release of silver ions. To date, it is still debatable as to whether any biological impact of nanosized silver is relative to either its size, or to its ionic constitution. The aim of this review therefore is to provide a comprehensive, interdisciplinary overview--for biologists, chemists, toxicologists as well as physicists--regarding the production of silver NPs, its (as well as in their ionic form) chemical and biochemical behaviours towards/within a multitude of relative and realistic biological environments and also how such interactions may be correlated across a plethora of different biological organisms.


Personality characteristics of young adult offspring of substance abusers: a study highlighting methodological issues.
Our study sought to demonstrate the importance of attending to methodological issues in the study of personality characteristics of offspring of substance abusers (OSAs). A 4 x 2 factorial design, Parental Mental Health (no known psychological problems, substance abuse, other psychological problems, or dual diagnosis) x Exposure to Abuse/Neglect (present or absent), was used to examine depression proneness, neuroticism, alexithymia, self-esteem, self-concept, and locus of control in young adults. Results of this study demonstrated that in order to determine whether characteristics of OSAs are uniquely related to parental substance abuse, a psychiatric control group is needed. Also, the importance of separating the influences of other family environment variables (e.g., exposure to abuse/neglect) was demonstrated. None of the differences between groups in this study could be attributed to parental substance abuse per se; rather, the concomitants of substance abuse (i.e., exposure to abuse/neglect) were found to be more strongly related to poorer adjustment in OSAs. Finally, it was shown that the proportion of treatment-seeking subjects included in the sample affected the level of pathology observed. These results are discussed in relation to previous findings, and recommendations regarding methodology are made for future studies.


Detection of circulating antigens in the diagnosis of acute toxoplasmosis.
Using an enzyme-linked immunosorbent assay, we have found circulating antigens of Toxoplasma gondii in three models of murine toxoplasmosis: mice infected with trophozoites of the RH strain (acute toxoplasmosis), the Beverley strain (subacute toxoplasmosis), and the T626 strain (chronic toxoplasmosis). Circulating antigens were detected 48 hr after infection in the mice infected with the RH strain, and all mice had antigenemia by the fourth day. In those infected with the Beverley strain, circulating antigens were detected from the second day after inoculation until the end of the study, with a peak (71% of the infected mice) on day 10. Of those infected with the T626 strain, 40% had antigenemia at 13 days after infection. The detection of circulating antigens in serum is directly related to the presence of toxoplasmosis in the acute phase in the three models studied and, therefore, may prove very useful in the rapid diagnosis of this disease.


The role of myocardial viability in contemporary cardiac practice.
Ischemic heart disease (IHD) remains the single most common cause of death worldwide. Ischemic cardiomyopathy is a major sequel of coronary artery disease. The economic health burden of IHD is substantial. In patients with old myocardial infarction (OMI), the extent of viable myocardium (VM) directly affects the short- and long-term outcome. There is a considerable collection of observational data showing substantial improvement in patients with significant left ventricular dysfunction when the need for revascularization is guided by preoperative assessment of viability and hibernation. However, a major challenge for present cardiovascular imaging is to identify better ways to assess viable but inadequately perfused myocardium and thus optimize selection of patients for coronary revascularization. Several non-invasive techniques have been developed to detect signs of viability. Hence, our aim is to provide the reader a state-of-the art review for the assessment of myocardial viability.


Surface Curvature Differentially Regulates Stem Cell Migration and Differentiation via Altered Attachment Morphology and Nuclear Deformation.
Signals from the microenvironment around a cell are known to influence cell behavior. Material properties, such as biochemical composition and substrate stiffness, are today accepted as significant regulators of stem cell fate. The knowledge of how cell behavior is influenced by 3D geometric cues is, however, strongly limited despite its potential relevance for the understanding of tissue regenerative processes and the design of biomaterials. Here, the role of surface curvature on the migratory and differentiation behavior of human mesenchymal stem cells (hMSCs) has been investigated on 3D surfaces with well-defined geometric features produced by stereolithography. Time lapse microscopy reveals a significant increase of cell migration speed on concave spherical compared to convex spherical structures and flat surfaces resulting from an upward-lift of the cell body due to cytoskeletal forces. On convex surfaces, cytoskeletal forces lead to substantial nuclear deformation, increase lamin-A levels and promote osteogenic differentiation. The findings of this study demonstrate a so far missing link between 3D surface curvature and hMSC behavior. This will not only help to better understand the role of extracellular matrix architecture in health and disease but also give new insights in how 3D geometries can be used as a cell-instructive material parameter in the field of biomaterial-guided tissue regeneration.


Distance Education among Italian Teachers: Differences and Experiences.
The successful integration of technology in teaching is a key component of education. Although prior research highlighted factors fostering the use of technology by teachers, few studies focused on whether these factors vary among teachers of different grade levels and subjects. Moreover, no studies have investigated personal experiences related to distance education among a large sample of teachers. To address these gaps, the present mixed-method study sought to examine whether factors promoting distance education varied among Italian teachers of different grade levels and subjects. A further aim was to explore experiences of teachers using distance education. The sample involved 357 Italian teachers and preservice teachers who completed an online questionnaire during the COVID-19 pandemic that also contained open-ended questions. Findings indicated that teaching self-efficacy was greater in pre-service and primary teachers, while facilitating conditions were greater in humanities and science secondary teachers. The perceived ease of use of technology and technology for pedagogy skills were more pronounced among science secondary teachers. Advanced technology skills were lower in humanities secondary teachers while the behavioural intention to use technology was greatest among pre-service teachers. Four themes emerged from the qualitative study of teachers' insights. These included positive and negative aspects of using technology, the relationship with students, the versatility of distance education, and the quality of lessons. This study underscores the need to address training based on different teachers' grade levels and subjects, and to focus on the emerging themes to better integrate the use of technology in schools.


A Systematic Approach To Promoting Home Hemodialysis during End Stage Kidney Disease.
Home dialysis has garnered much attention since the advent of the Advancing American Kidney Health initiative. For many patients and nephrologists, home dialysis and peritoneal dialysis are synonymous. However, home hemodialysis (HHD) should not be forgotten. Since 2004, HHD has grown more rapidly than other dialytic modalities. The cardinal feature of HHD is customizability of treatment intensity, which can be titrated to address the vexing problems of volume and pressure loading during interdialytic gaps and ultrafiltration intensity during each hemodialysis session. Growing HHD utilization requires commitment to introducing patients to the modality throughout the course of ESKD. In this article, we describe a set of strategies for introducing HHD concepts and equipment. First, patients initiating dialysis may attend a transitional care unit, which offers an educational program about all dialytic modalities during 3-5 weeks of in-facility hemodialysis, possibly using HHD equipment. Second, prevalent patients on hemodialysis may participate in "trial-run" programs, which allow patients to experience increased treatment frequency and HHD equipment for several weeks, but without the overt commitment of initiating HHD training. In both models, perceived barriers to HHD-including fear of equipment, anxiety about self-cannulation, catheter dependence, and the absence of a care partner-can be addressed in a supportive setting. Third, patients on peritoneal dialysis who are nearing a transition to hemodialysis may be encouraged to consider a home-to-home transition (i.e., from peritoneal dialysis to HHD). Taken together, these strategies represent a systematic approach to growing HHD utilization in multiple phenotypes of patients on dialysis. With the feature of facilitating intensive hemodialysis, HHD can be a key not only to satiating demand for home dialysis, but also to improving the health of patients on dialysis.


Tissue interactions in the regulation of axon pathfinding during tooth morphogenesis.
Like many other organs, the tooth develops as a result of the epithelial-mesenchymal interactions. In addition, the tooth is a well-defined peripheral target organ for sensory trigeminal nerves, which are required for the function and protection of the teeth. Dental trigeminal axon growth and patterning are tightly linked with advancing tooth morphogenesis and cell differentiation. This review summarizes recent findings on the regulation of dental axon pathfinding, which have provided evidence that the development of tooth trigeminal innervation is controlled by epithelial-mesenchymal interactions. The early dental epithelium possesses the information to instruct tooth nerve supply, and signals mediating these interactions are part of the signaling networks regulating tooth morphogenesis. Tissue interactions, thus, appear to provide a central mechanism of spatiotemporally orchestrating tooth formation and dental axon navigation and patterning.


Design of smart neonatal health monitoring system using SMCC.
Automated health monitoring and alert system development is a demanding research area today. Most of the currently available monitoring and controlling medical devices are wired which limits freeness of working environment. Wireless sensor network (WSN) is a better alternative in such an environment. Neonatal intensive care unit is used to take care of sick and premature neonates. Hypothermia is an independent risk factor for neonatal mortality and morbidity. To prevent it an automated monitoring system is required. In this Letter, an automated neonatal health monitoring system is designed using sensor mobile cloud computing (SMCC). SMCC is based on WSN and MCC. In the authors' system temperature sensor, acceleration sensor and heart rate measurement sensor are used to monitor body temperature, acceleration due to body movement and heart rate of neonates. The sensor data are stored inside the cloud. The health person continuously monitors and accesses these data through the mobile device using an Android Application for neonatal monitoring. When an abnormal situation arises, an alert is generated in the mobile device of the health person. By alerting health professional using such an automated system, early care is provided to the affected babies and the probability of recovery is increased.


An "unexpected" fatal case of the hypereosinophilic syndrome.
An "unexpected" fatal case of the hypereosinophilic syndrome in a 30-year-old man with perennial rhinitis, alcoholic intolerance and asthma is presented. Histological post-mortem examination showed extensive myocarditis and eosinophilic pneumonia.


IL-13/IL-4 signaling contributes to fibrotic progression of the myeloproliferative neoplasms.
Myelofibrosis (MF) is a disease associated with high unmet medical needs because allogeneic stem cell transplantation is not an option for most patients, and JAK inhibitors are generally effective for only 2 to 3 years and do not delay disease progression. MF is characterized by dysplastic megakaryocytic hyperplasia and progression to fulminant disease, which is associated with progressively increasing marrow fibrosis. Despite evidence that the inflammatory milieu in MF contributes to disease progression, the specific factors that promote megakaryocyte growth are poorly understood. Here, we analyzed changes in the cytokine profiles of MF mouse models before and after the development of fibrosis, coupled with the analysis of bone marrow populations using single-cell RNA sequencing. We found high interleukin 13 (IL-13) levels in the bone marrow of MF mice. IL-13 promoted the growth of mutant megakaryocytes and induced surface expression of transforming growth factor β and collagen biosynthesis. Similarly, analysis of samples from patients with MF revealed elevated levels of IL-13 in the plasma and increased IL-13 receptor expression in marrow megakaryocytes. In vivo, IL-13 overexpression promoted disease progression, whereas reducing IL-13/IL-4 signaling reduced several features of the disease, including fibrosis. Finally, we observed an increase in the number of marrow T cells and mast cells, which are known sources of IL-13. Together, our data demonstrate that IL-13 is involved in disease progression in MF and that inhibition of the IL-13/IL-4 signaling pathway might serve as a novel therapeutic target to treat MF.


Redundant nerve roots of the cauda equina. A case report.
One further case of redundant nerve roots of the cauda equina is described, and a review of eleven comparable cases in the literature is provided. In the authors' opinion, this disease appears to be a congenital anomaly, frequently but not necessarily associated with spinal changes.


Renal collecting duct carcinoma in a dog.
An 8-year-old, male, mongrel dog developed severe cough and anorexia and died within 3 months. Autopsy revealed an invasive grayish-white mass in the right kidney and multiple nodules in the lungs, thoracic wall, and spleen. Histologically, the renal mass and the other nodules were mainly composed of papillotubular structures lined by oval-to-polygonal pleomorphic cells. The cells were reactive with DBA, PNA, and UEA-1 lectins and positive for vimentin but negative for CD10 and high molecular weight cytokeratin. Because of its histological, histochemical, and immunohistochemical similarities with human collecting duct carcinoma (CDC), a diagnosis of renal collecting duct carcinoma with pulmonary, thoracic, and splenic metastases was established. To our knowledge, this is the first case report of CDC in animals.


High Potency of Melaleuca alternifolia Essential Oil against Multi-Drug Resistant Gram-Negative Bacteria and Methicillin-Resistant Staphylococcus aureus.
Herein, an extended investigation of Tea tree oil (TTO) against a number of multi-drug resistant (MDR) microorganisms in liquid and vapor phases is reported. The activity of TTO was tested against methicillin-sensitive Staphylococcus aureus (MSSA), Escherichia coli, and clinical strains of methicillin-resistant S. aureus (MRSA), extended-spectrum beta lactamases producer carbapenem-sensitive Klebsiella pneumoniae (ESBL-CS-Kp), carbapenem-resistant K. pneumoniae (CR-Kp), Acinetobacter baumannii (CR-Ab), and Pseudomonas aeruginosa (CR-Pa). Minimal inhibitory/bactericidal concentrations (MIC/MBCs) and synergistic activity between TTO and different antimicrobials were determined. In the vapor assay (VP), TTO-impregnated discs were placed on the lid of a petri dish and incubated for 24 h at 37 °C. TTO showed a potent bactericidal activity against all the tested microorganisms. TTO in combination with each reference antimicrobial showed a high level of synergism at sub-inhibitory concentrations, particularly with oxacillin (OXA) against MRSA. The VP assay showed high activity of TTO against CR-Ab. Evaluation of in-vitro activity clearly indicated TTO as a potential effective antimicrobial treatment either alone or in association with known drugs against MDR. Therefore, TTO could represent the basis for a possible role in non-conventional regimens against S. aureus and Gram-negative MDR. TTO in VP might represent a promising option for local therapy of pneumonia caused by CR-Ab.


Phaeosphamides A and B, Cytotoxic Cyclodecadepsipeptides from the Mangrove-Derived Fungus Phaeosphaeriopsis sp. S296.
Chemical examination of the fermented broth of the mangrove-derived fungus Phaeosphaeriopsis sp. S296 resulted in the isolation of two new cyclodecadepsipeptides, namely phaeosphamides A (1) and B (2), as well as one known congener Sch 217048 (3). The structures of new metabolites, including absolute configurations, were established on the basis of extensive spectroscopic data analyses, chemical conversion, and Marfey's method. The 2-hydroxy-3-methylpentanoic acid (Hmp) moiety and pipecolic acid (Pip) unit in structures were rarely discovered in nature. Interestingly, compounds 1-3 are examples of peptides discovered from the fungal genus Phaeosphaeriopsis for the first time. All identified compounds were evaluated for their cytotoxicity against five tumor cell lines of AGS, BEL-7402, HepG2, B16, and BIU87. Among them, compound 1 showed inhibitory activities against these tumor cell lines with IC50 values ranging from 5.14 to 66.38 μM. A further mechanistic investigation found that 1 arrested AGS cells in the G2 phase and induced their apoptosis in a dose-dependent manner.


IL-8-Positive Tumor-Infiltrating Inflammatory Cells Are a Novel Prognostic Marker in Pancreatic Ductal Adenocarcinoma Patients.
Tumor-infiltrating inflammatory cells (TIICs) in pancreatic ductal adenocarcinoma (PDAC) are reported to initiate and exacerbate invasion and metastasis. Interleukin-8 (IL-8), a proinflammatory cytokine, is expressed in both neoplastic cells and TIICs in PDAC tissues and increased in patient serum. The aim of this study is to evaluate the values of IL-8 expression profiles in tumor tissues and predict the source of serum IL-8 in PDAC patients. We used 2 independent groups of PDAC patient samples that included 240 cases. Tissue expression profiles of cytokines were evaluated with immunohistochemistry and serum levels with human IL-8 assay. The prognostic values of the variables were assessed by Kaplan-Meier or Cox regression analysis. Higher levels of IL-8-positive TIICs but not tumor cells in PDAC patients correlated with worse prognosis (P = 0.009) and higher blood serum IL-8 levels (P = 0.002). Controlling other independent factors, the relative hazard ratio for PDAC with higher IL-8-positive TIIC levels compared with those with lower TIIC levels was 1.588 (95% confidence interval, 1.04-2.42). Higher IL-8-positive TIIC levels in PDAC tumors indicate poorer prognosis and positively correlate with serum IL-8 concentrations and vice versa. These data suggested that IL-8 might have a potential target for PDAC therapies.


'Learn from the people you meet'.
Dawn Jarvis qualified as an enrolled nurse in 1987, converting to a registered nurse in 1993 and then a registered sick children's nurse in 1995. She has held many clinical and managerial roles as a children's nurse, including lead nurse in a paediatric high dependency unit, and programme director for children's and maternity services. She is now nursing programme lead for NHS England.


Amplitude modulation detection and temporal modulation cutoff frequency in normal hearing infants.
The goal of this study was to determine if temporal modulation cutoff frequency was mature in three-month-old infants. Normal-hearing infants and young adults were tested in a single-interval forced-choice observer-based psychoacoustic procedure. Two parameters of the temporal modulation transfer function (TMTF) were estimated to separate temporal resolution from amplitude modulation sensitivity. The modulation detection threshold (MDT) of a broadband noise amplitude modulated at 10 Hz estimated the y-intercept of the TMTF. The cutoff frequency of the TMTF, measured at a modulation depth 4 dB greater than the MDT, provided an estimate of temporal resolution. MDT was obtained in 27 of 33 infants while both MDT and cutoff frequency was obtained in 15 infants and in 16 of 16 adults. Mean MDT was approximately 10 dB poorer in infants compared to adults. In contrast, mean temporal modulation cutoff frequency did not differ significantly between age groups. These results suggest that temporal resolution is mature, on average, by three months of age in normal hearing children despite immature sensitivity to amplitude modulation. The temporal modulation cutoff frequency approach used here may be a feasible way to examine development of temporal resolution in young listeners with markedly immature sensitivity to amplitude modulation.


Gastric Aspergillosis in a COVID-19 Patient.
The SARS-CoV-2 virus, which causes Coronavirus-19 infection (COVID-19), frequently elicits the development of depressed immunity, therefore, opportunistic infections. Opportunistic organisms are commonly present in the human body without causing critical illness. However, they can also lead to pathologic illness when a person is immunocompromised. Aspergillosis is among the many opportunistic infections. Even though this infection primarily involves the respiratory system and is less likely to be found in the gastrointestinal tract, we report a case of a COVID-19 individual that developed massive gastrointestinal bleeding whose condition deteriorated, and the pathological examination revealed gastric aspergillosis. Although not common, gastric aspergillosis should be considered while treating patients with COVID-19 who present gastrointestinal symptoms.


A pharmaceutical monitoring system to assess the quality of antituberculosis drug products used in Mauritania.
The quality of drug products may be affected from manufacture to dispensing, particularly at high temperature and humidity as in Mauritania. This country is not included in the World Health Organization reports on poor quality products due to the lack of a qualified laboratory and monitoring system. Ensuring the quality of medicine is even more relevant in the case of diseases such as Tuberculosis, due to its high prevalence, complex treatment and continuous bacterial resistance. The aim was to develop a monitoring system to assess the quality of antituberculosis drugs products, by the substandard detection based on European and United States Pharmacopeial recommendations regarding quality control. In addition to studying the influence of accelerated storage conditions (40 ± 2°C/75 ± 5% relative humidity) on their qualities and comparing the dissolution profiles to contrast the quality. 18 antituberculosis drug products were taken from Europe and Mauritania, and quality was studied through visual inspection and according to the compliance of the mass uniformity, uniformity of dosage units, dissolution, disintegration and friability pharmacopeial tests. Furthermore, a dissolution profile comparison was carried out to examine quality. A stability study was conducted to assess the influence of climatic conditions on the content and the dissolved amount of the active pharmaceutical ingredients, which were determined by an ultra-performance liquid chromatography system. As result, 69.3% of 13 Mauritanian formulations had a substandard quality mainly due to non-compliance with the test for friability or content uniformity of these medicines. All European drug products complied with pharmacopeia specifications. In addition, storage conditions affected the dissolution rate of ethambutol and the uniformity of the 4 antituberculosis combination drug products.


Microbiological Characteristics of Carbapenem-Resistant Enterobacteriaceae Clinical Isolates Collected from County Hospitals.
To investigate the molecular characteristics of carbapenem-resistant Enterobacteriaceae (CRE) from county hospitals in China. Forty-three sequential non-duplicate CRE strains (including 33 Klebsiella pneumoniae isolates, 4 Enterobacter cloacae isolates, 3 Escherichia coli isolates, 1 Serratia marcescens, 1 Morganella morganii and 1 Citrobacter freundii) were collected from 4 county hospitals and 2 municipal hospitals. Antimicrobial susceptibility testing was conducted by broth microdilution method, using 3-aminophenylboronic acid and EDTA and the modified carbapenem inactivation method (mCIM) to screen phenotype of carbapenemase. β-Lactamases were characterized by polymerase chain reaction (PCR) and DNA sequencing. The transferability of bla NDM-5 was investigated by transformation experiment. Clonal relatedness was evaluated by pulsed-field gel electrophoresis and multilocus sequence typing . The results of antimicrobial susceptibility testing indicated that 43 CRE strains were resistant to most of the antimicrobial agents, except tigecycline and colistin. Overall, 93%, 93%, and 97.7% of these strains were resistant to imipenem, meropenem, and ertapenem, respectively. PCR and DNA sequencing indicated that 67.4% (29/43) were bla KPC-2 positive isolates, in which 3.4% (1/29) was coproduced with bla NDM-1. In addition, 7.0% (3/43), 4.7% (2/43), 4.7% (2/43), 2.3% (1/43), 2.3% (1/43) were bla NDM-1, bla NDM-16, bla NDM-4, bla NDM-5, bla IMP-4 positive isolates, respectively. The 29 bla KPC-2-positive isolates belonged to 12 different PFGE type and designated as ST11 (n=20) and ST15, ST39, ST116, ST667, ST2245, ST2338. The plasmid bearing bla NDM-5 could be transferred into recipient E. coli J53 through transformation. Our study indicated the dissemination of CRE between the tertiary hospitals and secondary hospitals.


[Mental suffering IN patients with parkinsonism].
Comprehensive treatment of patients with parkinson's disease demands that not only motoric dysfunctions but also psychosocial aspects have to be considered. The scope of this review is to cover not only the frequent dementia and depressive states associated with this disease, but also the drug-induced behavioral abnormalities, insomnia, conflicts with those providing care and implications for professional activities. The fundamental pathophysiological and psychopathological mechanisms are explained and illustrated by examples.


Breast cancer colonization by Fusobacterium nucleatum accelerates tumor growth and metastatic progression.
Fusobacterium nucleatum is an oral anaerobe recently found to be prevalent in human colorectal cancer (CRC) where it is associated with poor treatment outcome. In mice, hematogenous F. nucleatum can colonize CRC tissue using its lectin Fap2, which attaches to tumor-displayed Gal-GalNAc. Here, we show that Gal-GalNAc levels increase as human breast cancer progresses, and that occurrence of F. nucleatum gDNA in breast cancer samples correlates with high Gal-GalNAc levels. We demonstrate Fap2-dependent binding of the bacterium to breast cancer samples, which is inhibited by GalNAc. Intravascularly inoculated Fap2-expressing F. nucleatum ATCC 23726 specifically colonize mice mammary tumors, whereas Fap2-deficient bacteria are impaired in tumor colonization. Inoculation with F. nucleatum suppresses accumulation of tumor infiltrating T cells and promotes tumor growth and metastatic progression, the latter two of which can be counteracted by antibiotic treatment. Thus, targeting F. nucleatum or Fap2 might be beneficial during treatment of breast cancer.


Closing the loop.
The dream of closing the loop is actually the dream of creating an artificial pancreas and freeing the patients from being involved with the care of their own diabetes. Insulin-dependent diabetes (type 1) is a chronic incurable disease which requires constant therapy without the possibility of any 'holidays' or insulin-free days. It means that patients have to inject insulin every day of their life, several times per day, and in order to do it safely they also have to measure their blood glucose levels several times per day. Patients need to plan their meals, their physical activities and their insulin regime - there is only very small room for spontaneous activities. This is why the desire for an artificial pancreas is so strong despite the fact that it will not cure the diabetic patients. Attempts to develop a closed-loop system started in the 1960s but never got to a clinical practical stage of development. In recent years the availability of continuous glucose sensors revived those efforts and stimulated the clinician and researchers to believe that closing the loop might be possible nowadays. Many papers have been published over the years describing several different ideas on how to close the loop. Most of the suggested systems have a sensing arm that measures the blood glucose repeatedly or continuously, an insulin delivery arm that injects insulin upon command and a computer that makes the decisions of when and how much insulin to deliver. The differences between the various published systems in the literature are mainly in their control algorithms. However, there are also differences related to the method and site of glucose measurement and insulin delivery. SC glucose measurements and insulin delivery are the most studied option but other combinations of insulin measurements and glucose delivery including intravascular and intraperitoneal (IP) are explored. We tried to select recent publications that we believe had influenced and inspired people interested in the field.


On the mechanisms that limit oxygen uptake during exercise in acute and chronic hypoxia: role of muscle mass.
Peak aerobic power in humans (VO2,peak) is markedly affected by inspired O2 tension (FIO2). The question to be answered in this study is what factor plays a major role in the limitation of muscle peak VO2 in hypoxia: arterial O2 partial pressure (Pa,O2) or O2 content (Ca,O2)? Thus, cardiac output (dye dilution with Cardio-green), leg blood flow (thermodilution), intra-arterial blood pressure and femoral arterial-to-venous differences in blood gases were determined in nine lowlanders studied during incremental exercise using a large (two-legged cycle ergometer exercise: Bike) and a small (one-legged knee extension exercise: Knee)muscle mass in normoxia, acute hypoxia (AH) (FIO2 = 0.105) and after 9 weeks of residence at 5260 m (CH). Reducing the size of the active muscle mass blunted by 62% the effect of hypoxia on VO2,peak in AH and abolished completely the effect of hypoxia on VO2,peak after altitude acclimatization. Acclimatization improved Bike peak exercise Pa,O2 from 34 +/- 1 in AH to 45 +/- 1 mmHg in CH(P <0.05) and Knee Pa,O2 from 38 +/- 1 to 55 +/- 2 mmHg(P <0.05). Peak cardiac output and leg blood flow were reduced in hypoxia only during Bike. Acute hypoxia resulted in reduction of systemic O2 delivery (46 and 21%) and leg O2 delivery (47 and 26%) during Bike and Knee, respectively, almost matching the corresponding reduction in VO2,peak. Altitude acclimatization restored fully peak systemic and leg O(2) delivery in CH (2.69 +/- 0.27 and 1.28 +/- 0.11 l min(-1), respectively) to sea level values (2.65 +/- 0.15 and 1.16 +/- 0.11 l min(-1), respectively) during Knee, but not during Bike. During Knee in CH, leg oxygen delivery was similar to normoxia and, therefore, also VO2,peak in spite of a Pa,O2 of 55 mmHg. Reducing the size of the active mass improves pulmonary gas exchange during hypoxic exercise, attenuates the Bohr effect on oxygen uploading at the lungs and preserves sea level convective O2 transport to the active muscles. Thus, the altitude-acclimatized human has potentially a similar exercising capacity as at sea level when the exercise model allows for an adequate oxygen delivery (blood flow x Ca,O2), with only a minor role of Pa,O2 per se, when Pa,O2 is more than 55 mmHg.


Screening of hepatocyte proteins binding to F protein of hepatitis C virus by yeast two-hybrid system.
To investigate the biological function of F protein by yeast two-hybrid system. We constructed F protein bait plasmid by cloning the gene of F protein into pGBKT7, then recombinant plasmid DNA was transformed into yeast AH109 (a type). The transformed yeast AH109 was mated with yeast Y187 (alpha type) containing liver cDNA library plasmid in 2XYPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-alpha-gal for selection and screening. After extracting and sequencing plasmids from positive (blue) colonies, we underwent sequence analysis by bioinformatics. Thirty-six colonies were selected and sequenced. Among them, 11 colonies were zymogen granule protein, 5 colonies were zinc finger protein, 4 colonies were zinc-alpha-2-glycoprotein, 1 colony was sialyltransferase, 1 colony was complement control protein factor I, 1 colony was vitronectin, and 2 colonies were new genes with unknown function. The yeast two-hybrid system is an effective method for identifying hepatocyte proteins interacting with F protein of hepatitis C virus. F protein may bind to different proteins.


[Determination of biological activity of therapeutic muds].
Different neurophysiological healing mud effect on laboratory animals was determined. The effect of healing mud application depends on type, chemical composition and nature of healing mud. The technique applied in the study may be used to test biological activity of peloids in a complex of physiological studies.


Developmental acquisition of DNase I sensitivity of the phosphoenolpyruvate carboxykinase (GTP) gene in rat liver.
The sensitivity to DNase I digestion of the gene encoding rat phosphoenolpyruvate carboxykinase (GTP) (EC 188.8.131.52) was assessed during development and prior to the onset of expression. This gene is resistant to DNase I digestion in nuclei isolated from livers of 19-day rat fetuses. Gradual acquisition of sensitivity of the phosphoenolpyruvate carboxykinase gene, which starts later than the 19th day of gestation and is completed by the 21st day, occurs before initiation of gene expression. As transcription of the phosphoenolpyruvate carboxykinase gene is not detected until birth, the events observed may represent a shift from a dormant to an active gene. Injection of N6,O2-dibutyryladenosine 3',5'-cyclic monophosphate into fetuses on the 19th day of gestation induces gene expression and sensitivity to DNase I digestion within 3 hr of treatment. While this short treatment does not affect the methylation pattern of the gene, longer treatment of fetuses (2 days) with dibutyryl-cAMP results in premature hypomethylation of the gene. A hierarchy of modifications of the phosphoenolpyruvate carboxykinase gene during development is discussed.


A network view on psychiatric disorders: network clusters of symptoms as elementary syndromes of psychopathology.
The vast number of psychopathological syndromes that can be observed in clinical practice can be described in terms of a limited number of elementary syndromes that are differentially expressed. Previous attempts to identify elementary syndromes have shown limitations that have slowed progress in the taxonomy of psychiatric disorders. To examine the ability of network community detection (NCD) to identify elementary syndromes of psychopathology and move beyond the limitations of current classification methods in psychiatry. 192 patients with unselected mental disorders were tested on the Comprehensive Psychopathological Rating Scale (CPRS). Principal component analysis (PCA) was performed on the bootstrapped correlation matrix of symptom scores to extract the principal component structure (PCS). An undirected and weighted network graph was constructed from the same matrix. Network community structure (NCS) was optimized using a previously published technique. In the optimal network structure, network clusters showed a 89% match with principal components of psychopathology. Some 6 network clusters were found, including "Depression", "Mania", "Anxiety", "Psychosis", "Retardation", and "Behavioral Disorganization". Network metrics were used to quantify the continuities between the elementary syndromes. We present the first comprehensive network graph of psychopathology that is free from the biases of previous classifications: a 'Psychopathology Web'. Clusters within this network represent elementary syndromes that are connected via a limited number of bridge symptoms. Many problems of previous classifications can be overcome by using a network approach to psychopathology.


Adaptation to an Intracellular Lifestyle by a Nitrogen-Fixing, Heterocyst-Forming Cyanobacterial Endosymbiont of a Diatom.
The symbiosis between the diatom Hemiaulus hauckii and the heterocyst-forming cyanobacterium Richelia intracellularis makes an important contribution to new production in the world's oceans, but its study is limited by short-term survival in the laboratory. In this symbiosis, R. intracellularis fixes atmospheric dinitrogen in the heterocyst and provides H. hauckii with fixed nitrogen. Here, we conducted an electron microscopy study of H. hauckii and found that the filaments of the R. intracellularis symbiont, typically composed of one terminal heterocyst and three or four vegetative cells, are located in the diatom's cytoplasm not enclosed by a host membrane. A second prokaryotic cell was also detected in the cytoplasm of H. hauckii, but observations were infrequent. The heterocysts of R. intracellularis differ from those of free-living heterocyst-forming cyanobacteria in that the specific components of the heterocyst envelope seem to be located in the periplasmic space instead of outside the outer membrane. This specialized arrangement of the heterocyst envelope and a possible association of the cyanobacterium with oxygen-respiring mitochondria may be important for protection of the nitrogen-fixing enzyme, nitrogenase, from photosynthetically produced oxygen. The cell envelope of the vegetative cells of R. intracellularis contained numerous membrane vesicles that resemble the outer-inner membrane vesicles of Gram-negative bacteria. These vesicles can export cytoplasmic material from the bacterial cell and, therefore, may represent a vehicle for transfer of fixed nitrogen from R. intracellularis to the diatom's cytoplasm. The specific morphological features of R. intracellularis described here, together with its known streamlined genome, likely represent specific adaptations of this cyanobacterium to an intracellular lifestyle.


Simultaneous Monitoring of Multi-Enzyme Activity and Concentration in Tumor Using a Triply Labeled Fluorescent In Vivo Imaging Probe.
The use of fluorescent imaging probes that monitor the activity of proteases that experience an increase in expression and activity in tumors is well established. These probes can be conjugated to nanoparticles of iron oxide, creating a multimodal probe serving as both a magnetic resonance imaging (MRI) agent and an indicator of local protease activity. Previous works describe probes for cathepsin D (CatD) and metalloproteinase-2 (MMP2) protease activity grafted to cross-linked iron oxide nanoparticles (CLIO). Herein, we have synthesized a triply labeled fluorescent iron oxide nanoparticle molecular imaging (MI) probe, including an AF750 substrate concentration reporter along with probes for cathepsin B (CatB) sand MMP2 protease activity. The reporter provides a baseline signal from which to compare the activity of the two proteases. The activity of the MI probe was verified through incubation with the proteases and tested in vitro using the human HT29 tumor cell line and in vivo using female nude mice injected with HT29 cells. We found the MI probe had the appropriate specificity to the activity of their respective proteases, and the reporter dye did not activate when incubated in the presence of only MMP2 and CatB. Probe fluorescent activity was confirmed in vitro, and reporter signal activation was also noted. The fluorescent activity was also visible in vivo, with injected HT29 cells exhibiting fluorescence, distinguishing them from the rest of the animal. The reporter signal was also observable in vivo, which allowed the signal intensities of the protease probes to be corrected; this is a unique feature of this MI probe design.


[Clinical-pathomorphologic preparation of patients after surgical treatment of differentiated thyroid cancer to adjuvant treatment with 131I].
Thyroid cancer treatment includes: surgery, radioiodine therapy, thyroxine therapy and radiotherapy. Selection of treatment's strategy depends on histopathological evaluation, age of patient and iodine uptake. The aim of the analysis was to verify how the medical documentation of patients treated by surgical treatment was prepared. All patients were directed to the Department of Endocrinology, Holycross Cancer Center in Kielce. Analysis encompassed 33 patients classified to 131I therapy for the first time. In each case patient's medical documentation and histopathological diagnosis made in Department of Tumor Pathology, Holycross Cancer Center, were compared. A conformity with primary cancer diagnosis was obtained in all but one patient in whom pathological consultation did not confirm oxyphilic carcinoma. In four cases the type of cancer was changed as a result of repeated consultation. pTMN classification was stated in 23 primary pathologic examination cases. After consultation of slides and inspection of surgery protocol, total or partial pTNM stage was obtained in other 9 patients. Establishment of diagnostic and therapeutic algorithm, accepted on Conference in Szczyrk, 1995 is inadequately executed in small centers. Routine consultation of histopathological slides creates a possibility to make a proper choice of treatment's strategy.


Regio- and diastereoselective preparation of tetrahydrobenzo[c]-1-aza-2lambda(5)-phospholes through dearomatization cyclization of lithiated N-benzyl-N-alkyl(diphenyl)phosphinamides. Synthesis of gamma-(N-alkylamino)phosphinic acids.
A study of the protonation of the cycloadducts derived from the dearomatization reaction of lithiated N-alkyl-N-benzyldiphenylphosphinamides has been carried out. The regio- and stereoselectivity of the process has been analyzed in terms of the size of the N-alkyl substituent, the acidity and size of the protonating reagent, and the cosolvent used. The optimization of these variables allowed the preparation of tetrahydrobenzo[c]-1-aza-2lambda(5)-phospholes containing a 1,3-cyclohexadiene or 1,4-cyclohexadiene system with moderate to excellent regio- and stereocontrol. The heterocycles were readily hydrolyzed, affording gamma-(N-alkylamino)diphenylphosphinic acids with the functionalities linked to a cyclohexadiene substructure.


Feasibility, Safety, and Utility of Advanced Critical Care Transesophageal Echocardiography Performed by Pulmonary/Critical Care Fellows in a Medical ICU.
Competence in advanced critical care transesophageal echocardiography allows the intensivist to diagnose and manage cardiopulmonary failure when transthoracic echocardiography is not sufficient to answer the clinical question. This report describes the feasibility, safety, and utility of transesophageal echocardiography performed by pulmonary and critical care fellows in a medical ICU in critically ill patients who were mechanically ventilated. This longitudinal descriptive study of advanced transesophageal echocardiography was performed in patients who were mechanically ventilated. The complications and clinical utility of the examinations were abstracted from review of the image set, the written report, and the patient chart. Influence on diagnosis was classified as confirming the initial clinical diagnosis or identification of a new or unanticipated finding. Therapeutic impact was categorized as no change in therapy resulting from the study findings or as change in therapy resulting from the study findings. The examination was feasible in all patients in whom it was attempted, and there were no complications of the procedure. New or actionable findings were identified in 43 (28%) studies, and 109 (72%) studies confirmed the initial clinical diagnosis. The findings resulted in a change in clinical management in 58 (38%) patients. Critical care transesophageal echocardiography is feasible, safe, and has clinical utility. It can be safely and effectively performed by fellows within the context of their critical care training with faculty supervision. Pulmonary critical care fellowship training programs should consider introducing critical care transesophageal echocardiography as a useful clinical tool.


Isolation and molecular characterization of mRNA transport mutants in Schizosaccharomyces pombe.
Nucleocytoplasmic transport of mRNA is essential for eukaryotic gene expression. However, how mRNA is exported from the nucleus is mostly unknown. To elucidate the mechanisms of mRNA transport, we took a genetic approach to identify genes, the products of which play a role in that process. From about 1000 temperature -sensitive (ts- or cs-) mutants, we identified five ts- mutants that are defective in poly(A)+ RNA transport by using a situ hybridization with an oligo(dT)50 as a probe. These mutants accumulate poly(A)+ RNA in the nuclei when shifted to a nonpermissive temperature. All five mutations are tightly linked to the ts- growth defects, are recessive, and fall into four different groups designated as ptr 1-4 (poly(A)+ RNA transport). Interestingly, each group of mutants has a differential localization pattern of poly(A)+ RNA in the nuclei at the nonpermissive temperature, suggesting that they have defects at different steps of the mRNA transport pathway. Localization of a nucleoplasmin-green fluorescent protein fusion suggests that ptr2 and ptr3 have defects also in nuclear protein import. Among the isolated mutants, only ptr2 showed a defect in pre-mRNA splicing. We cloned the ptr2+ and ptr3+ genes and found that they encode Schizosaccharomyces pombe homologues of the mammalian RCC1, a guanine nucleotide exchange factor for RAN/TC4, and the ubiquitin-activating enzyme E1 involved in ubiquitin conjugation, respectively. The ptr3+ gene is essential for cell viability, and Ptr3p tagged with green fluorescent protein was localized in both the nucleus and the cytoplasm. This is the first report suggesting that the ubiquitin system plays a role in mRNA export.


[Cryptococcal meningitis successfully treated with liposomal amphotericin B and voriconazole in an elderly patient].
Abstract An 81-year-old woman who had microscopic polyangiitis that was being treated with corticosteroids for 2 months was admitted to our department because of fever and clouding of consciousness. Neurological examination showed disturbance of consciousness and nuchal stiffness. Analysis of cerebrospinal fluid (CSF) revealed pleocytosis, low glucose level, and elevated protein levels. On the basis of the presence of Cryptococcus neoformans in CSF, the patient was diagnosed with cryptococcal meningitis. On the basis of established practice guidelines, liposomal amphotericin B (L-AMB) was administered to avoid the possible nephrotoxicity of amphotericin B. After the treatment was started, the patient's condition gradually improved. The results of CSF analysis also showed a gradual recovery. Because the cryptococcal antigen in CSF did not disappear completely, voriconazole (VRCZ) was administered orally; subsequently, the CSF cryptococcal antigen titer gradually decreased. During the course of the treatment with L-AMB and VRCZ, there were no severe side effects that required a change in treatment. To the best of our knowledge, in Japan, the combination of L-AMB and VRCZ has rarely been reported to be effective for the treatment of cryptococcal meningitis. The recovery of our patient indicates that the administration of L-AMB and VRCZ to elderly patients with cryptococcal meningitis and renal insufficiency is safe and leads to a successful outcome.


[Long-term toxicity test of arginine esterase from Agkistrodon halys ussuriensis venom].
To study the long-term toxicity of arginine esterase from Agkistrodon halys ussuriensis venom for the clinical application of arginine esterase in the future. Beagle dogs were used as experimental animals and were divided into control group, arginine esterase low dose group (0.06 u/kg), the middle dose group (0.18 u/kg) and high dose group (0.36 u/kg). Every group consisted of four dogs. The arginine esterase was given intravenously once a day for 180 days. Then three dogs in each group were sacrificed and the fourth one was fed without injecting arginine esterase for 15 days. The toxic reactions during treatment and recovery period were determined by evaluating and comparing the general criteria ( including locomotor activity, growth rate, appetite and death rate), clinical criteria (including blood test and urine test), pathological dissection and viscera coefficient of the treated animals and the control animals. There were no significant differences in general criteria. The clinical criteria of the treated animals were the same as those of the control animals except liver function. There were no significant differences in pathological dissection and viscera coefficient between the treated animals and the control animals except livers. The livers in high dose arginine esterase treated animals were swollen and vacuolated and there was significant difference in liver coefficient between them (P<0.05). The toxic symptom of liver disappeared after withdrawal of treatment. From these results, the non-toxic dose of arginine esterase for dogs was estimated to be 0.18 u/kg under the present study conditions and is about 15 times the clinical dosage for using the drug "Qingshuanmei" of which the main component is arginine esterase. The long-term toxicity test result indicates that the toxicity of pure arginine esterase is lower than that of "Qingshuanmei", suggesting that clinical use of the arginine esterase is safe.


Defining and improving response to treatment in patients with bipolar disorder.
Functional outcomes are more meaningful measures of response to treatment for bipolar disorder than are scores on various psychiatric rating scales (all of which have limitations) used to gauge improvement in symptoms. With the former approach, patients are considered to be in remission if they achieve normal or near-normal levels of functioning in occupational, family, and social settings. Sleep patterns are reliable indicators of whether a patient with bipolar disorder is likely to relapse or sustain remission in the near term. Regularly scheduled nightly sleep periods may help prevent rapid cycling in patients with mania, while perturbations in circadian rhythms may be early markers of impending relapse. Medications used to attain response and/or remission in maintenance therapy include lithium and valproate. The choice of mood stabilizer depends on the patient's symptoms, prior response to a mood stabilizer, and tolerance of the drug. For patients requiring additional therapy, combination regimens with mood stabilizers and atypical antipsychotics appear effective. Psychoeducation for patients and families and interpersonal psychotherapy also can help prolong remission.


Prevalence and effectiveness of laxative use among elderly residents in a regional hospital affiliated nursing home in hsinchu county.
Long-term care residents are susceptible to constipation and one-half to three quarter of older nursing home residents receive laxatives regularly. The purpose of this study was to evaluate the factors related to abnormal bowel function and explore the effectiveness of laxative treatment among the elderly residents of a nursing home. A total of 110 residents older than 65 years old was enrolled in this study. The following variables were gathered: age, gender, body mass index (BMI), length of stay, daily fluid intake, type of food, functional level, cognitive ability, physical therapy status, somatic and psychiatric diseases, number of medications, and medication use. The use and dosage of laxatives were recorded by means of Anatomical Therapeutic Chemical (ATC) classification system. Normal bowel function was defined as defecation frequency from three defecations per day to three defecations per week and stool consistency score of three to five on Bristol Stool Form Scale. A comparison between groups with normal and abnormal bowel function was drawn. Low BMI, increased fluid intake, liquid food intake, poor functional level, poor cognition, and a history of stroke were significantly associated with altered bowel function (P < 0.05). The most frequently used laxatives were glycerol, senna glycoside, and magnesium oxide. There were significant differences in laxative regimens between residents with normal and altered bowel function; those with altered bowel function tended to take more laxatives than those with normal bowel function. This study suggested that treatment of constipation in the nursing home was unsatisfactory. To improve treatment outcomes in those susceptible to altered bowel function, a coordinated approach with involvement of physicians, nursing staff, and other professionals including dieticians and pharmacists seems necessary.


Application of direct covalent molecular assembly in the fabrication of polyimide ultrathin films.
Ultrathin films were fabricated using synthesized hydroxyl polyimide (HPI) in a layer-by-layer fashion on amine-terminated substrates of silicon, quartz, and gold. The interlayer linkages were established by using terephthaloyl chloride as a bridging agent to form ester groups between HPI layers. Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, UV-vis absorption spectroscopy, atomic force microscopy, ellipsometry, and electrochemical impedance spectroscopy were employed to study the interfacial chemistry, stepwise growth, morphology, thickness, optical property, and insulation behavior of the assembled film. The films show excellent stability and strength, which can be attributed to the covalent interlayer linkage.


[Design of heterologous mitochondria: import of cattle cytochrome P-450SCC precursors in plant mitochondria].
It has been shown that pre-P-450scc of bovine adrenal cortex mitochondria synthesized in a rabbit reticulocyte lysate cell-free system, is translocated into isolated soybean cotyledon mitochondria, thereby taking the mature form size. This finding is suggestive of the occurrence of a specific receptor and maturase for pre-P-450scc in plant mitochondria. Thus, plant mitochondria can be used as recipients for the mammalian cholesterol hydroxylase system in an attempt to study the mechanism of its formation and preservation.


Germ cell differentiation from pluripotent cells.
Infertility is a medical condition with an increasing impact in Western societies with causes linked to toxins, genetics, and aging (primarily delay of motherhood). Within the different pathologies that can lead to infertility, poor quality or reduced quantity of gametes plays an important role. Gamete donation and therefore demand on donated sperm and eggs in fertility clinics is increasing. It is hoped that a better understanding of the conditions related to poor gamete quality may allow scientists to design rational treatments. However, to date, relatively little is known about human germ cell development in large part due to the inaccessibility of human development to molecular genetic analysis. It is hoped that pluripotent human embryonic stem cells and induced pluripotent stem cells may provide an accessible in vitro model to study germline development; these cells are able to differentiate to cells of all three primary embryonic germ layers, as well as to germ cells in vitro. We review the state of the art in germline differentiation from pluripotent stem cells.


Outcomes of the Conventional versus Pocket-Creation Method for Endoscopic Submucosal Dissection of Gastric Body Tumors Using a Dual Knife: A Retrospective Study.
Various endoscopic submucosal dissection (ESD) methods for gastric tumors have been tried. However, no studies have yet compared results according to the ESD method for gastric body tumors using a dual knife. The objective of this study was to compare outcomes of two ESD methods for gastric body tumors: the pocket-creation method and conventional method. Patients who underwent ESD for a gastric body tumor were retrospectively reviewed. Patients were divided into two groups according to the ESD method: the conventional method (group I) and pocket-creation method (group II). Characteristics of patients and tumors, hospitalization period, incidence of complications, resection margin status, incidence of surgical operation, procedure time, and laboratory findings were investigated. Of the total of 100 patients, 52 belonged to group I and 48 to group II. All tumors were successfully resected en bloc. Resection margin involvement was found in six (11.5%) of group I and six (12.5%) of group II. Complications were observed in seven (13.5%; major complication five, minor two) of group I and eight (16.7%; major two, minor six) of group II. There were no significant differences in ESD outcomes such as hospitalization period, incidence of complications, resection margin status, incidence of surgical operation, procedure time, or inflammatory response after ESD between the two groups. Both methods are suitable for treating gastric body tumors with adequate treatment success rates and comparable complication rates.


Hybrid Polymer-Silica Nanostructured Materials for Environmental Remediation.
Due to the ever-growing global population, it is necessary to develop highly effective processes that minimize the impact of human activities and consumption on the environment. The levels of organic and inorganic contaminants have rapidly increased in recent years, posing a threat to ecosystems. Removing these toxic pollutants from the environment is a challenging task that requires physical, chemical, and biological methods. An effective solution involves the use of novel engineered materials, such as silica-based nanostructured materials, which exhibit a high removal capacity for various pollutants. The starting materials are also thermally and mechanically stable, allowing for easy design and development at the nanoscale through versatile functionalization procedures, enabling their effective use in pollutant capture. However, improvements concerning mechanical properties or applicability for repeated cycles may be required to refine their structural features. This review focuses on hybrid/composite polymer-silica nanostructured materials. The state of the art in nanomaterial synthesis, different techniques of functionalization, and polymer grafting are described. Furthermore, it explores the application of polymer-modified nanostructured materials for the capture of heavy metals, dyes, hydrocarbons and petroleum derivatives, drugs, and other organic compounds. The paper concludes by offering recommendations for future research aimed at advancing the application of polymer-silica nanostructured materials in the efficiency of pollutant uptake.


Mutation Screening of the CHCHD2 Gene for Alzheimer's Disease and Frontotemporal Dementia in Chinese Mainland Population.
As an important multifunctional protein involved in regulation of mitochondrial metabolism, CHCHD2 was identified as a causative gene for Parkinson's disease (PD), yet the relationship between CHCHD2 and neurodegenerative dementia is not well understood. We directly sequenced the entire coding region of CHCHD2 gene in 150 AD patients, 84 FTD patients, and 417 controls. Four rare putative pathogenic variants of CHCHD2, including rs142444896 (c.5C>T, p.P2L), rs752705344 (c.15C>G, p.S5R), rs145190179 (c.94G>A, p.A32T), and rs182992574 (c.255T>A, p.S85R) were identified from a cohort composed of 150 AD and 84 FTD patients. These results suggest that CH CHD2 gene play an important role in other neurodegenerative disorders from our dementia study in China.


Endotoxin, tumor necrosis factor, and related mediators: new approaches to shock.
For some time, it has been clear that septic shock results from the coordinated release of cytokine mediators that directly or indirectly injure tissues of the host. Natural and synthetic inhibitors of tumor necrosis factor, interleukin-1, and other key cytokines may alter current concepts regarding shock and its therapy.


The development of a nonintrusive home-based physiologic signal measurement system.
This paper describes the development of a nonintrusive health-monitoring house system that has been developed for monitoring patients' health status. The system functions unobtrusively and leaves patients free to conduct daily activities. Installed sensors and devices in the house monitor biosignals such as bedside electrocardiogram (ECG), snoring, weight, and movement pattern. The network in the house was constructed using asymmetric digital subscriber line (ADSL) and Bluetooth technologies and it is capable of transmitting monitored signals to the remote hospital server through the home server. The system is found to be effective in monitoring patients' daily activity and health status in a nonconstraining manner.


A new rapid and sensitive bioluminescence assay for antibiotics that inhibit protein synthesis.
A new sensitive, rapid and simple bioluminescence assay for antibiotics inhibiting protein synthesis is described. In this assay the ability of the tested antibiotic to inhibit the de novo synthesis of the enzymes participating in the bacterial luminescence system is determined by means of a dark variant of a luminous bacterium that undergoes prompt induction of the luminescence system with certain DNA-intercalating agents. Upon induction, the in vivo luminescence of the dark variant is increased more than 50-fold within 30 min. Antibiotics that block the de novo synthesis of protein limit the development of luminescence at a level that was found to be a function of the antibiotic concentration. The minimum detectable concentration of antibiotics in the bioluminescence test, after 45-60 min of incubation, was 0.1 microgram/ml for streptomycin, gentamicin, kanamycin, lincomycin and chloramphenicol and 0.3 microgram/ml for neomycin, clindamycin and spectinomycin. The new bioluminescence test has been used to assay these antibiotics in serum.


Patterns of surgical treatment for women diagnosed with early breast cancer in Queensland.
Australian women with early breast cancer should be given the choice between breast-conserving surgery (BCS) or mastectomy. This is the first Australian study to report on patterns of surgical care specifically for early breast cancer at a population level. Two population-based routine data collections were linked to obtain surgical treatment information for breast cancer cases diagnosed in 2004 in Queensland, from which we identified 1274 cases of early female breast cancer. Logistic regression was used to assess the likelihood of female breast cancer patients having mastectomy, BCS, and axillary node dissection, after adjusting for patient and hospital demographics, tumor size, and comorbidities. Three-quarters (77%) of women had BCS, 29% had a mastectomy, and 86% had dissection of the axillary lymph nodes. The likelihood of women having mastectomy was higher among women living in rural areas, those treated in public hospitals, and women who had comorbidities of anemia or heart failure. In contrast, BCS was more likely for women treated in private hospitals or hospitals with high surgical caseload. Heart failure decreased the likelihood of BCS. Having an axillary node dissection was more likely among younger women and those treated in high caseload hospitals. The observed differentials in surgical treatment for early breast cancer patients suggest that access issues may have contributed to the decision-making process. Understanding the reasons why women with early breast cancer choose a certain treatment strategy should be a focus of future research.


Road-runoff metal concentrations in Toledo, Ohio, and their relation to average daily traffic and age of pavement overlay.
Road runoff can contain metal concentrations and sampling of it can be expensive if auto-samplers are used. In this study, an inexpensive storm water sampling apparatus was developed and used for collecting the road runoff at eleven different sites in Toledo area. Sites were selected with the purpose of evaluating the effects of average daily traffic (ADT) and the age of pavement overlay on road runoff metal concentrations. Ag, As, Cd, and Mo concentrations were primarily below detection limit. Ba, Mn, Ni, Cu, Pb, and Zn were observed in higher concentrations but lower than <800 mug/L. Mg, Al, Fe, K, Na, and Ca were observed in mg/L levels, much higher concentrations. ADT was positively correlated with runoff concentrations of K, Mn, Pb and Zn. The source of Ca, Mg, and Mn appeared to be the asphalt pavement surface; with time, as the pavement aged, these sources appeared to be depleted as observed by lower runoff concentrations in older pavements.


Mitochondrial Effects of Teucrium Polium and Prosopis Farcta Extracts in Colorectal Cancer Cells
Background: Teucrium Polium and Prosopis Farcta have been traditionally employed in cancer treatment. In this study we evaluated the effects of methanolic extracts of these two plants in HT-29 cells. Methods: IC50s of extracts were obtained via MTT assay and the levels of ROS production, cell death, collapse of mitochondrial membrane potential and Sirt3 enzyme activity were determined. Results: After 48 hours exposure, IC50s for Teucrium and Prosopis extracts were 3 and 2μg/ml, respectively. Extracts induced higher ROS production after 6 hours than after 12 hours. Mitochondrial membrane potential collapse and cell death rate were also increased; Teucrium caused greater cell death than Prosopis. Extracts from both plants increased Sirt3 activity in its normal form, but only Teucrium extract caused a significant increase in activity of Sirt3 enzyme isolated from cancer cells. Conclusion: Teucrium and Prosopis extracts exert anticancer activity via mitochondrial alterations, as exemplified by increased ROS levels, Sirt3 activity and cell death in HT-29 colorectal cancer cells.


Cushing's Disease Presenting with Functional Neurological (Conversion) Disorder.
While psychiatric manifestations are common in patients with Cushing's syndrome (CS), to our knowledge, there are no reported cases of CS presenting with functional neurological disorder (FND), a neuropsychiatric condition in which patients experience neurological symptoms, such as motor dysfunctions, sensory symptoms, speech disorders, or nonepileptic seizures, in the absence of neurological disease. Here, we report a case of a complex patient with Cushing's disease who presented with multiple FND symptoms including nonepileptic seizures, bilateral lower extremity paralysis, decreased finger flexion resulting in limited hand function, and stuttering. This case illustrates a rare psychiatric manifestation of CS presenting as multiple neurological complaints. Furthermore, we elucidate how a multidisciplinary treatment approach improved our patient's FND symptoms.


Assessment of QT-prolonging drugs in the isolated normal and failing rabbit hearts.
Lengthening of QTc is the usual signal to indicate torsadogenic potential of a therapeutic agent. The ICH S7B guideline recommends that new chemical entities should be assessed for potential of delayed ventricular repolarization in animal models. The aim of this study was to determine a feasibility of using isolated failing heart rabbit to assess the QT-lengthening drugs in comparison with their effects on isolated normal heart rabbits. Heart failure was induced by ligation of the left anterior descending and descending branch of left circumflex coronary arteries. One month after ligation, all rabbits were anesthetized and the hearts were removed quickly, and they were perfused with the oxygenated Krebs-Henseleit solution to which escalating concentrations of QT-lengthening compounds were added. RR, QT, and QTc(F) were not significantly different, at rest, between failing and normal hearts. During baseline, dP/dt<inf>max</inf> was lower and dP/dt<inf>min</inf> was higher for failing hearts than for normals. In responses to all three QT-lengthening compounds, RR, QT and QTc(F) lengthened similarly in a dose-response manner in both the failing and normal hearts. Neither the failing nor the normal hearts developed fatal arrhythmias, torsades de pointes. Langendorff preparations of failing hearts are as good as normal isolated hearts and can be use to assess the potential of delayed ventricular repolarization of test articles.


SUMO protease SENP1 deSUMOylates and stabilizes c-Myc.
Posttranslational modifications play a crucial role in the proper control of c-Myc protein stability and activity. c-Myc can be modified by small ubiquitin-like modifier (SUMO). However, how SUMOylation regulates c-Myc stability and activity remains to be elucidated. The deSUMOylation enzyme, SENP1, has recently been shown to have a prooncogenic role in cancer; however, mechanistic understanding of this is limited. Here we show that SENP1 is a c-Myc deSUMOylating enzyme. SENP1 interacts with and deSUMOylates c-Myc in cells and in vitro. Overexpression of wild-type SENP1, but not its catalytically inactive C603S mutant, markedly stabilizes c-Myc and increases its levels and activity. Knockdown of SENP1 reduces c-Myc levels, induces cell cycle arrest, and drastically suppresses cell proliferation. We further show that c-Myc can be comodified by both ubiquitination and SUMOylation. SENP1-mediated deSUMOylation reduces c-Myc polyubiquitination, suggesting that SUMOylation promotes c-Myc degradation through the proteasome system. Interestingly, SENP1-mediated deSUMOylation promotes the accumulation of monoubiquitinated c-Myc and its phosphorylation at serine 62 and threonine 58. SENP1 is frequently overexpressed, correlating with the high expression of c-Myc, in breast cancer tissues. Together, these results reveal that SENP1 is a crucial c-Myc deSUMOylating enzyme that positively regulates c-Myc's stability and activity.


Acoustic and articulatory correlates of stop consonants in a parrot and a human subject.
General acoustic patterns in productions of American English stops /p,b,t,d,k,g/ by a Grey parrot (Psittacus erithacus) and his primary human trainer are identified. Comparisons demonstrated both differences (coherence of voicing [/p,t,k/ and /b,d,g/] and place [/p,b/, /t,d/, and /k,g/] subsets, predictive power of measures related to F1 and F3) and similarities (categorical distinctions among stop consonants) in acoustic properties of psittacine and human speech. The authors suggest how acoustic data might be correlated with articulatory events in the parrot. Our data are also compared with speech from a mynah (Gracula religiosa). Because humans perceive and correctly label psittacine stop consonants despite differences in avian and human articulatory and acoustic parameters, our findings (a) further expand our initial work with vowels [Patterson and Pepperberg, J. Acoust. Soc. Am. 96, 634-648 (1994)], (b) corroborate findings suggesting that the perceptual discontinuities exploited by speech may be accessible to various sensory systems, and (c) challenge the notion that human speech production is a unique process.


Dynamic gene expression profiling using a microfabricated living cell array.
We describe the development of a microfluidic platform for continuous monitoring of gene expression in live cells. This optically transparent microfluidic device integrates high-throughput molecular stimulation with nondestructive monitoring of expression events in individual living cells, hence, a living cell array (LCA). Several concentrations of a soluble molecular stimulus are generated in an upstream microfluidic network and used to stimulate downstream reporter cells, each containing a green fluorescence reporter plasmid for a gene of interest. Cellular fluorescence is continuously monitored and quantified to infer the expression dynamics of the gene being studied. We demonstrate this approach by profiling the activation of the transcription factor NF-kappaB in HeLa S3 cells in response to varying doses of the inflammatory cytokine TNF-alpha. The LCA platform offers a unique opportunity to simultaneously control dynamic inputs and measure dynamic outputs from adherent mammalian cells in a high-throughput fashion. This approach to profiling expression dynamics, in conjunction with complementary techniques such as DNA microarrays, will help provide a more complete picture of the dynamic cellular response to diverse soluble stimuli.


Generation of MLC-2v-tdTomato knock-in reporter mouse line.
MLC-2v is a myosin light chain regulatory protein which is specifically expressed in ventricular cardiomyocytes and slow twitch skeletal muscle cells. MLC-2v plays critical roles in ventricular maturation during heart development. Mice lacking MLC-2v are embryonic lethal due to heart failure associated with abnormal myofibrillar organization of ventricular cardiomyocytes. To study the development of ventricular cardiac muscle and slow twitch skeletal muscle, we generated a new MLC-2v reporter mouse line by knocking-in a tdTomato reporter cassette into 3' UTR of the MLC-2v gene without disrupting the endogenous gene. Our results demonstrated specific MLC-2v-tdTomato knock-in reporter expression in ventricular cardiomyocytes and slow twitch muscle during myogenesis, precisely recapitulating the spatiotemporal expression pattern of endogenous MLC-2v. No tdTomato expression was observed in the atria, fast twitch muscle or other organs throughout development into adulthood. Isolated neonatal and adult ventricular cardiomyocytes uniformly express tdTomato. Taken together, MLC-2v-tdTomato knock-in reporter mouse model described in this article will serve as a valuable tool to study cardiac chamber and skeletal muscle specification during development and regeneration by overcoming the pitfalls of transgenic strategies.