"Real-world" practical evaluation strategies: a review of telehealth evaluation.
Currently, the increasing interest in telehealth and significant technological breakthroughs of the past decade create favorable conditions for the widespread adoption of telehealth services. Therefore, expectations are high that telehealth can help alleviate prevailing challenges in health care delivery. However, in order to translate current research to policy and facilitate adoption by patients and health care providers, there is need for compelling evidence of the effectiveness of telehealth interventions. Such evidence is gathered from rigorously designed research studies, which may not always be practical in many real-world settings. Our aim was to summarize current telehealth evaluation strategies and challenges and to outline practical approaches to conduct evaluation in real-world settings using one of our previously reported telehealth initiatives, the Diabetes Connect program, as a case study. We reviewed commonly used current evaluation frameworks and strategies, as well as best practices based on successful evaluative efforts to date to address commonly encountered challenges in telehealth evaluation. These challenges in telehealth evaluation and commonly used frameworks are described relevant to the evaluation of Diabetes Connect, a 12-month Web-based blood glucose monitoring program. Designers of telehealth evaluation frameworks must give careful consideration to the elements of planning, implementation, and impact assessment of interventions. Evaluating performance at each of these phases is critical to the overall success of an intervention. Although impact assessment occurs at the end of a program, our review shows that it should begin at the point of problem definition. Critical to the success of an evaluative strategy is early planning that involves all stakeholders to identify the overall goals of the program and key measures of success at each phase of the program life cycle. This strategy should enable selection of an appropriate evaluation strategy and measures to aid in the ongoing development and implementation of telehealth and provide better evidence of program impact. We recommend a pragmatic, multi-method, multi-phase approach to telehealth evaluation that is flexible and can be adapted to the characteristics and challenges unique to each telehealth program.


Bis(4-eth-oxy-phen-yl) sulfoxide.
In the title compound, C(16)H(18)O(3)S, the dihedral angle between the benzene rings is 82.7 (2)°. The O atom of the sulfoxide group is disordered over two orientations with refined occupancy factors of 0.563 (3):0.437 (3). In the crystal, mol-ecules are linked by inter-molecular C-H⋯O hydrogen bonds, forming chains along the b axis.


[The locked-in syndrome].
The locked-in syndrome is poorly understood, and may be misdiagnosed as coma. The syndrome consists of complete paralysis of all four limbs and the lower cranial nerve pairs, associated with an unaltered consciousness. Vertical eye movements and blinking remain intact, so allowing some form of communication by way of eye codes. Obstruction of the vertebral and basilar vascular system is the major cause, but the six cases reported show the aetiological diversity of this syndrome. If the vascular obstruction is diagnosed within the first few hours, fibrinolytic therapy may be considered. Although the patients usually die within a few days or after several months, some rare cases of recovery after several weeks have been reported. Intensive care is required by these patients (tracheostomy, artificial ventilation, intensive nursing care); the physicians and nursing staff looking after them should always keep in mind that these patients are conscious and able to communicate.


Stress monitoring through non-invasive instrumental analysis of skin conductivity.
This paper envelops the notion of yogic breathing and its applications as one of the techniques for stress management through regulation of skin conductivity among Indian engineering students. Stress perturbs the normal physiological or psychological functioning of an individual. Thus, it is extremely vital to manage stress. To explore the consequence of yogic breathing on skin conductivity using galvanic skin response sensor meter (GSRSM) on engineering students, of different universities. The study uses stratified random sampling technique for the experimental study undertaken to measure skin conductivity. 471 Engineering students, in the age group of 18-22 years who gave informed written consent were screened, of which 71 (15%) dropped out and 400 students (84.9%) completed the study. The GSRSM was used to record skin conductivity (pre and post) among experimental (n = 200) and Control group (n = 200) each comprising of 52 females (26%) and 148 males (74%). The Experimental group underwent yogic breathing whereas the Control group did not do so. The experimental group reported reduction in mean value in skin conductivity after deep breathing for the time period (t = 300 sec) as compared to the control group, and this was significant (p < 0.01 and p < 0.05). It is recommended that, this uncomplicated and yet extremely effectual ancient technique of yogic breathing could be included in core curriculum to reduce and manage stress among engineering students. The paper thus highlights the use of yogic technique as an effective mode for Stress Management amongst Indian Engineering students.


[Prognostic factors in severe head injury].
The knowledge of the so called prognostic factors or indicators involved in severe head injury (SHI) is an issue of great interest to make predictions about the future of patients with this pathology. Those indicators constitute the basic elements of the different prognostic formulas or models carried out in order to make predictions in SHI. The mentioned models, therefore, will be constructed by a group of variables (prognostic indicators or factors) and several scales (prognostic scales) that are useful for measuring the final outcome of these patients. In this paper we resume, after an exhaustive review of the literature, the knowledge about the prognostic factors related to SHI. These indicators have been classified as follows: clinical, radiological, physiological, and biochemical. Moreover, we have briefly described the prognostic scales more commonly used in SHI.


Investigation of the milling capabilities of the F10 Fine Grind mill using Box-Behnken designs.
Size reduction or milling of the active is often the first processing step in the design of a dosage form. The ability of a mill to convert coarse crystals into the target size and size distribution efficiently is highly desirable as the quality of the final pharmaceutical product after processing is often still dependent on the dimensional attributes of its component constituents. The F10 Fine Grind mill is a mechanical impact mill designed to produce unimodal mid-size particles by utilizing a single-pass two-stage size reduction process for fine grinding of raw materials needed in secondary processing. Box-Behnken designs were used to investigate the effects of various mill variables (impeller, blower and feeder speeds and screen aperture size) on the milling of coarse crystals. Response variables included the particle size parameters (D10, D50 and D90), span and milling rate. Milled particles in the size range of 5-200 μm, with D50 ranging from 15 to 60 μm, were produced. The impeller and feeder speeds were the most critical factors influencing the particle size and milling rate, respectively. Size distributions of milled particles were better described by their goodness-of-fit to a log-normal distribution (i.e. unimodality) rather than span. Milled particles with symmetrical unimodal distributions were obtained when the screen aperture size was close to the median diameter of coarse particles employed. The capacity for high throughput milling of particles to a mid-size range, which is intermediate between conventional mechanical impact mills and air jet mills, was demonstrated in the F10 mill. Prediction models from the Box-Behnken designs will aid in providing a better guide to the milling process and milled product characteristics.


Proton magnetic resonance spectroscopy in dementia of Alzheimer type.
Reduced N-acetyl aspartate (NAA) and increased myo-inositol (MI) levels have been reported in patients with dementia of Alzheimer type (DAT) in comparison with controls. We wished to assess the validity of these findings and to evaluate possible correlations of metabolite proportions with cognitive dysfunction in DAT. Twelve patients with DAT and 10 healthy age-matched controls were included. The severity of dementia was assessed using different scales including the Mini-Mental State Examination. MRS was performed with a conventional 1.5 Tesla scanner in a single voxel in the centrum semi-ovale (TE = 30 ms or TE = 136 ms; TR = 1500 ms). The evaluation of MRS results was limited by low interrater, intermeasurement (different echo times) and test-retest reliabilities, by a high interindividual variance and by the failure to measure absolute metabolite concentrations. These problems in mind, it was remarkable that previously reported reductions of NAA levels in patients with DAT could be reproduced in the present sample. The proportion of NAA was diminished in demented subjects in comparison with controls (37% vs 44.90%, short TE). A non-significant trend towards minor reductions of creatine, choline and MI proportions in these subjects might indicate that proportions of other metabolites necessarily increase when NAA is reduced. Cognitive dysfunction of demented subjects was significantly correlated with reductions of NAA, but not with increases of MI. Due to the present technical and methodological problems and to the non-specificity of findings, proton MRS cannot be applied to support the diagnosis of DAT in a clinical setting.


The applied anatomy of the thoracolumbar fascia.
The thoracolumbar fascia was studied by dissection in ten adult human cadavers. The posterior layer of this fascia was found to consist of two laminae. The superficial lamina is formed by the aponeurosis of latissimus dorsi. The deep lamina consists of bands of fibers passing caudolaterally from the midline. Both laminae form a retinaculum over the back muscles, and the deep lamina constitutes a series of accessory posterior ligaments that anchor the L2 to L5 spinous processes to the ilium and resist flexion of the lumbar spine. The function of these ligaments is enhanced by the contraction of the back muscles and the action of certain, restricted portions of the abdominal muscles.


[Electrophysiology of subthalamic nucleus in normal and Parkinson's disease].
Subthalamic nucleus (STN) has been known to play an important role in the regulation of cortico-basal ganglia-thalamo-cortical loop. STN neurons have pacemaking activitiy and their firing pattern can switch from spike mode to bursting mode when membrane potential becomes hyperpolarized. Recent study has shown that STN neurons show marked increase in burst and oscillatory activity during the dopamine-depleting state of Parkinson's disease (PD). This electrophysiological change in activity is now considered as an characterstic pathophysiological feature of PD. High frequency stimulation of STN can modify and "normalize" the activity of STN neurons in the pathophysiologial state. This electrophysiological treatment applied to STN, known as deep brain stimulation (DBS) clinically, ameliorates the symptoms of PD effectively, and is becoming a standard treatment in patients with advanced PD. This article would review the basic researches concerning electrical activities of STN and try to extend the basic knowledge into clinical applications.


Pharmaceutical perspective of neuropathic pain management for primary care providers.
Neuropathic pain (NP) is a chronic condition that affects ~ 1% of the general population globally. Several conditions such as chronic diabetes, herpes zoster (HZ), cancer, HIV, stroke, multiple sclerosis, physical compression or damage of nerves and certain surgical procedures can lead to neuropathy and related pain. The condition is difficult to treat with traditional analgesic drugs. Typically, non-traditional analgesics are used in treating pain in this condition such as antidepressants and antiepileptic drugs. Opioids are useful in some patients, but the risk of addiction and the risk of both short-term and long-term adverse effects make it a low priority drug class in the treatment of NP. In the current review we discuss the pharmacology and pharmaceutical care aspects of various classes of drugs used in the treatment of NP, counselling points for these drug classes, and future prospects in the treatment of NP.


Jury still out on extent of employers' cost savings under managed care. An AMA policy research perspective.
President Clinton's Health Security Act as well as other proposals for national health system reform and state-level reform call for greater use of managed care to control employers' health care costs. Even without government intervention, the trend is for increased enrollment of employees into managed care plans as opposed to traditional indemnity plans. In fact, the proportion of private employees enrolled in managed care plans grew from 5% in 1980 to 55% (90 million persons) in 1992. Despite the increased reliance on managed care plans, little is known about their ability to limit costs and whether they pass any savings on to employers. This American Medical Association Policy Research Perspectives summarizes a recent report by the General Accounting Office (GAO) that discusses their review of the literature on managed care and interviews with insurers and more than 60 private employers between March 1992 and March 1993.


Animal models in the neurotoxicology of 2,4-D.
2,4-D is a selective pre- and postemergence herbicide used for several crops. It is hazardous for the environment and risk for humans; therefore, several studies attempt to evaluate its effects and consequences of its use. The nervous system is supposedly a target for this herbicide, and this comprehensive review gathers the information about animal models that have been used for the study of the neurotoxicity of 2,4-D. The studies used several methods to evaluate the neurotoxicity of this herbicide, most of which used rodents, mainly rats, two used fish, and one used chicken eggs. The main behavioral effect observed concerned alterations in locomotor patterns and reduced motor activity. Biochemical analysis showed decreased levels of serotonin (5-HT) and increased levels of its metabolites and increased or decreased levels of DA and its metabolites depending on the brain area analyzed. Hypomyelination is also a possible effect of 2,4-D when the exposure occurs during the proliferation and development of the oligodendrocytes. The worst neuropathologic effects were observed in fish. Since most studies focused on the neurotoxicity of 2,4-D in rodents, the effect it may have on other species and groups of animals, especially with different physiology, is unclear and it should be researched.


The yin and yang of yeast transcription: elements of a global feedback system between metabolism and chromatin.
When grown in continuous culture, budding yeast cells tend to synchronize their respiratory activity to form a stable oscillation that percolates throughout cellular physiology and involves the majority of the protein-coding transcriptome. Oscillations in batch culture and at single cell level support the idea that these dynamics constitute a general growth principle. The precise molecular mechanisms and biological functions of the oscillation remain elusive. Fourier analysis of transcriptome time series datasets from two different oscillation periods (0.7 h and 5 h) reveals seven distinct co-expression clusters common to both systems (34% of all yeast ORF), which consolidate into two superclusters when correlated with a compilation of 1,327 unrelated transcriptome datasets. These superclusters encode for cell growth and anabolism during the phase of high, and mitochondrial growth, catabolism and stress response during the phase of low oxygen uptake. The promoters of each cluster are characterized by different nucleotide contents, promoter nucleosome configurations, and dependence on ATP-dependent nucleosome remodeling complexes. We show that the ATP:ADP ratio oscillates, compatible with alternating metabolic activity of the two superclusters and differential feedback on their transcription via activating (RSC) and repressive (Isw2) types of promoter structure remodeling. We propose a novel feedback mechanism, where the energetic state of the cell, reflected in the ATP:ADP ratio, gates the transcription of large, but functionally coherent groups of genes via differential effects of ATP-dependent nucleosome remodeling machineries. Besides providing a mechanistic hypothesis for the delayed negative feedback that results in the oscillatory phenotype, this mechanism may underpin the continuous adaptation of growth to environmental conditions.


Career Needs Assessment for Early Career Academic Surgeons Using a Modified Accelerated Delphi Process.
Over the past 2 decades, physicians' wellbeing has become a topic of interest. It is currently unclear what the current needs are of early career academic surgeons (ECAS). Consensus statements on academic needs were developed during a Delphi process, including all presenters from the previous European Surgical Association (ESA) meetings (2018-2022). The Delphi involved (1) a literature review, (2) Delphi form generation, and (3) an accelerated Delphi process. The Delphi form was generated by a steering group that discussed findings identified within the literature. The modified accelerated e-consensus approach included 3 rounds over a 4-week period. Consensus was defined as >80% agreement in any round. Forty respondents completed all 3 rounds of the Delphi. Median age was 37 years (interquartile range 5), and 53% were female. Majority were consultant/attending (52.5%), followed by PhD (22.5%), fellowship (15%), and residency (10%). ECAS was defined as a surgeon in 'development' years of clinical and academic practice relative to their career goals (87.9% agreement). Access to split academic and clinical contracts is desirable (87.5%). Consensus on the factors contributing to ECAS underperformance included: burnout (94.6%), lack of funding (80%), lack of mentorship (80%), and excessive clinical commitments (80%). Desirable factors to support ECAS development included: access to e-learning (90.9%), face-to-face networking opportunities (95%), support for research team development (100%), and specific formal mentorship (93.9%). The evolving role and responsibilities of ECAS require increasing strategic support, mentorship, and guidance on structured career planning. This will facilitate workforce sustainability in academic surgery in the future.


Realization of all logic gates and memory latch in the SC-CNN cell of the simple nonlinear MLC circuit.
We investigate the State-Controlled Cellular Neural Network framework of Murali-Lakshmanan-Chua circuit system subjected to two logical signals. By exploiting the attractors generated by this circuit in different regions of phase space, we show that the nonlinear circuit is capable of producing all the logic gates, namely, or, and, nor, nand, Ex-or, and Ex-nor gates, available in digital systems. Further, the circuit system emulates three-input gates and Set-Reset flip-flop logic as well. Moreover, all these logical elements and flip-flop are found to be tolerant to noise. These phenomena are also experimentally demonstrated. Thus, our investigation to realize all logic gates and memory latch in a nonlinear circuit system paves the way to replace or complement the existing technology with a limited number of hardware.


IMPROVEMENT OF COGNITIVE EFFICIENCY THROUGH COGNITIVE TRAINING IN HEALTHY SUBJECTS.
The aim of the study was to explore whether application of cognitive stimulation in young healthy subjects may improve their cognitive efficiency. The study included 12 healthy young subjects divided into two groups, experimental group and control group. Prior to cognitive stimulation treatment, both groups underwent baseline measurements with selected neuropsychological tests. The groups were matched with regard to the achievement on the baseline test. Only the experimental group underwent daily application of different computer-based cognitive tasks lasting for an hour a day for two weeks. After the treatment, both groups were tested with the same neuropsychological battery used at the baseline measurement. The experimental group showed a statistically significant difference between the measurements on the variables assessing immediate retention of visual material and recognition of verbal material. In addition, qualitative analysis showed that the experimental group also had better performance on the variables assessing delayed recall of visual material, visual and verbal range of attention, and delayed recall of verbal material. In conclusion, two-week cognitive stimulation in healthy subjects improves cognitive performance, expressed as higher average values of certain neuropsychological variables.


Subchronic Oral Dose Toxicity Study of Enterococcus Faecalis 2001 (EF 2001) in Mice.
As a part of general toxicity studies of Enterococcus Faecalis 2001 (EF 2001) prepared using heat-treatment bacillus mort body EF 2001 in mice, this study examined the toxicity of EF 2001 in single and repeated administrations following the previous report in order to apply this product to preventive medicine. The safety of oral ingestion of EF 2001 was examined in 6-week-old male and female ICR mice with 1,000 mg/kg, 3,000 mg/kg and 5,000 mg/kg body weight/day administrated by gavage of the maximum acceptable dose of EF 2001. The study was conducted using distilled water as a control following the methods for general toxicity studies described in the "Guidelines for Non-clinical Studies of Pharmaceutical Products 2002". As a control, 1) observation of general conditions, 2) measurement of body weight, 3) determination of food consumption, 4) determination of water consumption, 5) blood test and urinalysis and 6) pathological examination were performed for the administration of EF 2001. Mice received EF 2001 for 13 weeks and results were compared with those of the control group that received distilled water. The results of the above examinations revealed no significant differences between control and EF 2001 groups for both males and females. Thus, no notable toxicity was confirmed with single and repeated oral administrations of EF 2001. Oral administration in the above doses did not result in abnormal symptoms or death during the observation period. No abnormalities in blood cell count or organ weights were seen. Without any evidence of toxicity to cells and organs, EF 2001 is speculated to not adversely affect living organisms. The 50% lethal dose of EF 2001 with oral administration in mice is estimated to be greater than 5,000 mg/kg body weight/day for both male and female mice. Therefore, LD50 value for animals was 5,000 mg/kg or more.


How Social Status Shapes Person Perception and Evaluation: A Social Neuroscience Perspective.
Inferring the relative rank (i.e., status) of others is essential to navigating social hierarchies. A survey of the expanding social psychological and neuroscience literatures on status reveals a diversity of focuses (e.g., perceiver vs. agent), operationalizations (e.g., status as dominance vs. wealth), and methodologies (e.g., behavioral, neuroscientific). Accommodating this burgeoning literature on status in person perception, the present review offers a novel social neuroscientific framework that integrates existing work with theoretical clarity. This framework distinguishes between five key concepts: (1) strategic pathways to status acquisition for agents, (2) status antecedents (i.e., perceptual and knowledge-based cues that confer status rank), (3) status dimensions (i.e., domains in which an individual may be ranked, such as wealth), (4) status level (i.e., one's rank along a given dimension), and (5) the relative importance of a given status dimension, dependent on perceiver and context characteristics. Against the backdrop of this framework, we review multiple dimensions of status in the nonhuman and human primate literatures. We then review the behavioral and neuroscientific literatures on the consequences of perceived status for attention and evaluation. Finally, after proposing a social neuroscience framework, we highlight innovative directions for future social status research in social psychology and neuroscience.


THE GENETICS OF ASYMMETRICAL MALE STERILITY IN DROSOPHILA MOJAVENSIS AND DROSOPHILA ARIZONENSIS HYBRIDS: INTERACTIONS BETWEEN THE Y-CHROMOSOME AND AUTOSOMES.
Crosses between Drosophila mojavensis and D. arizonensis produce fertile females, but the males from the cross ♂ D. mojavensis × ♀ D. arizonensis are sterile. The chromosomal basis of sperm immotility was studied in these hybrids. Interspecific crossing-over was avoided by crossing hybrid males to pure-species females, and chromosomal identification in backcross progeny was possible by means of electrophoretic markers. The main findings are as follows. The Y-chromosome and two autosomes are involved in the determination of sperm motility. The other autosomes, with the exception of the sixth which was not tested, appear to have no effect. The effect of the D. arizonensis X-chromosome was not examined, but it is established that the D. mojavensis X-chromosome has no effect on sperm motility in males carrying the D. arizonensis Y-chromosome and any combination of autosomes. The Y-chromosome and the two autosomes interact with each other in a simple and predictable way, so that certain combinations of these chromosomes always produce motile sperm and others immotile sperm. Thus, asymmetrical male hybrid sterility may have a simple genetic basis. In contrast to ethological isolation, the genetic basis for this postmating isolating mechanism does not appear to vary among conspecific populations, an observation which suggests that postmating isolation antedates ethological isolation in these species.


["Psychological only children"--neuroses and illness behavior in adulthood].
Our first basic hypothesis concerning the increased " overindulgence " of the "only child" according to the psychological definition of the term (hereafter referred to as "only child") and his associated characteristic qualities, e. g. neurotic passiveness , attitudes of indolence and pretension , inclination toward substitute gratification, lack of control, and timid evasion, has not proved valid where our patients are concerned. The "only child" among our patients did not exhibit these qualities any more often than our other neurotic patients. In the male "only child" we observed a conspicuous psychopathologic accentuation with regard to his behaviour in personal attachments. Also, provided that age groupings are the same, he is more often unmarried than other male neurotic patients, and he exhibits patterns of contact and attachment which are characterized by suspicion and remoteness . There is no indication that the "only child" among our patients seeks medical advice earlier or with fewer mental reservations because of his being accustomed to take advantage of outside help. In contrast to expectation, the behaviour of the male "only child" with regard to accepting the assistance of others is characterized by extraordinary reserve. He is not reported ill as often as the other male neurotic patients nor does hospitalization for longer periods of time occur as frequently as with them. This is interpreted as an indication that the male "only child" is "healthier" than other neurotic patients and less frequently tends to inflict injuries upon himself. With regard to his qualifications in school and in his profession the "only child" is particularly advanced. The question is raised as to whether the literary stereotype of the " overindulged " only child is in need of a certain differentiation, at least with respect to the social background of the patient. Our "only-child" patients originate predominantly from lower social strata and do not exhibit the classical attitudes of overindulgence any more often than other neurotic patients. Possibly " overindulged " patients mainly descend from the upper social classes.


An update in diabetes management.
Staying abreast of the numerous recent advances in diabetes management presents a challenge to healthcare professionals. This paper provides an overview of these changes and discusses their implications for rehabilitation caregivers and clients. Because of increased scientific knowledge of the disease, there have been changes in nomenclature, diagnostic criteria, and nutritional guidelines. Medications to improve control of diabetes and innovations in equipment have been developed. The concepts of intensive management and self-management have gained importance, as have reimbursement issues. An overview of these advances and their implications for nursing practice, may help nurses throughout the continuum of care to assist individuals with diabetes to enjoy the maximum quality of life possible.


Cue Exposure Therapy for Alcohol Use Disorders: What Can Be Learned from Exposure Therapy for Anxiety Disorders?
Background: Cue Exposure Therapy (CET) has shown efficacy for treating alcohol use disorders (AUDs). Exposure therapy is highly effective for treating anxiety. Both techniques involve repeated, controlled exposures to alcohol or fear-related stimuli. Objectives: We considered the mechanisms of CET for AUDs by comparing it to exposure therapy for anxiety. Method: We conducted a narrative review contrasting theoretical literature examining the mechanisms of CET versus exposure therapy for anxiety. We reviewed RCTs and acute laboratory paradigms examining CET for AUDs. We considered common areas of emerging research, including the use of d-Cycloserine (DCS) and virtual reality (VR). Results: We found evidence that exposure therapy and CET at least partially achieve their effects through extinction learning. We found evidence that CET for AUDs is effective, with comparable benefits to other effective psychosocial treatments. DCS and VR have shown some limited success for augmenting CET for AUDs, so further research is needed to determine their value. Conclusions: There are theoretical and practical similarities between exposure to fear cues and cues of addiction, especially regarding extinction learning. However, these processes are also unique, particularly regarding the differing motivational properties of fear versus reward-related stimuli. We propose that unlike exposure for anxiety, CET takes effect by increasing self-control with each unreinforced exposure. We consider reasons for CET's limited use for AUDs, including its lower acceptability to clients and clinicians. We also note the limited evidence for CET for other substance use disorders, highlighting the need for continued investigation into its mechanisms and efficacy.


[Study of the action of x-ray contrast substances on the respiratory chain of rat liver mitochondria].
Subject to study was the influence of roentgen-contrast media on the respiration and linked with it phosphorylation in the mitochondria of the rats' liver. Triotrast, cardiotrast, bilignost and endografin are shown to produce no effect on the oxidative phosphorylation in concentrations that can be built up following their administration "in vivo", but inhibit the respiratory mitochondrial chain. Triombrin, urotrast and iodamide have no effect on the mitochondria.


Purification and Characterization of Recombinant N-Terminally Pyroglutamate-Modified Amyloid-β Variants and Structural Analysis by Solution NMR Spectroscopy.
Alzheimer's disease (AD) is the leading cause of dementia in the elderly and is characterized by memory loss and cognitive decline. Pathological hallmark of AD brains are intracellular neurofibrillary tangles and extracellular amyloid plaques. The major component of these plaques is the highly heterogeneous amyloid-β (Aβ) peptide, varying in length and modification. In recent years pyroglutamate-modified amyloid-β (pEAβ) peptides have increasingly moved into the focus since they have been described to be the predominant species of all N-terminally truncated Aβ. Compared to unmodified Aβ, pEAβ is known to show increased hydrophobicity, higher toxicity, faster aggregation and β-sheet stabilization and is more resistant to degradation. Nuclear magnetic resonance (NMR) spectroscopy is a particularly powerful method to investigate the conformations of pEAβ isoforms in solution and to study peptide/ligand interactions for drug development. However, biophysical characterization of pEAβ and comparison to its non-modified variant has so far been seriously hampered by the lack of highly pure recombinant and isotope-enriched protein. Here we present, to our knowledge, for the first time a reproducible protocol for the production of pEAβ from a recombinant precursor expressed in E. coli in natural isotope abundance as well as in uniformly [U-15N]- or [U-13C, 15N]-labeled form, with yields of up to 15 mg/l E. coli culture broth. The chemical state of the purified protein was evaluated by RP-HPLC and formation of pyroglutamate was verified by mass spectroscopy. The recombinant pyroglutamate-modified Aβ peptides showed characteristic sigmoidal aggregation kinetics as monitored by thioflavin-T assays. The quality and quantity of produced pEAβ40 and pEAβ42 allowed us to perform heteronuclear multidimensional NMR spectroscopy in solution and to sequence-specifically assign the backbone resonances under near-physiological conditions. Our results suggest that the presented method will be useful in obtaining cost-effective high-quality recombinant pEAβ40 and pEAβ42 for further physiological and biochemical studies.


[Predominantly intraductal low-grade differentiated adenocarcinoma in young man with long time mask clinic of chronic pancreatitis].
The authors present the case of diagnosis of low-grade adenocarcinoma of the pancreas, a man 34 years old, a long time to mask clinic chronic pancreatitis. Laboratory and instrumental investigations, including tumor markers CD 19-9, CEA, ultrasound, CT, MRI showed no reliable data for the presence of tumor. On the other hand, a young man of 34 years present credible evidence of chronic pancreatitis, in the form of a typical pain, a significant increase in serum am- ylase and lipase, and the presence of parenchymal and ductal signs of pancreatitis in visual research methods. Diagnosis of tumors, including obtaining morphological material, pre-operative pain relief were obtain by endoscopic methods. Histological examination of surgical specimens was detected a rare type of poorly differentiated adenocarcinoma of the pancreas tubulo-papillary structures, with predominant intraductal growth and a minimum depth of invasion due to pronounced fibrotic outcome variables, which leads to a complex disease diagnostics. After 1.5 years, after adequately performed surgery the patient was revealed metastases to the liver.


[Functional neuroimaging of the amygdala: the response to threatening and phobogenic stimuli].
Recent functional neuroimaging studies show that the amygdala has a central role in threat evaluation, in response to conditioned and unconditioned stimuli, in fear learning and fear extinction. The amygdala is involved in the pathophysiology of phobias and anxiety. In this review we critically examine the main findings of functional neuroimaging studies reporting data on the amygdala. Findings suggest that the response of the amygdala to threatening stimuli is mainly modulated by the infralimbic and prefrontal cortices, which inhibit activation of the amygdala (top-down inhibition), and by the hippocampus, the function of which is related to stimulus learning. The activity of the amygdala is modulated by various factors, like stimulus type and origin, emotion triggered by stimulus perception, and attention. The neural network comprising the amygdala and the frontal cortex is involved not only in top-down inhibition, but also in the emotional perception of facial expressions. This network also includes the thalamic pulvinar, which is densely interconnected with the amygdala, directly or indirectly, and which is activated by emotional face recognition of scary fear. Both top-down inhibition mechanisms and emotional face recognition are altered in anxiety disorders, particularly in specific and social phobia, resulting in reduced amygdalar activity inhibition after anxiety - or fear - inducing stimulus perception. Future functional neuroimaging studies will be able to provide new insights of normal and altered neurophysiology of the amygdala.


[Mitochondrial encephalomyopathies].
Mitochondrial Encephalomyopathies are primary disorders of energy metabolism recently described. They are the result of mitochondrial abnormalities with a wide spectrum of syndromes implying a multisystemic but predominantly muscular and cerebral involvement. Biochemical, histological, imagiologic and clinical features of the three well known mitochondrial syndromes (MERRF, MELAS and KSS) are described, as well as the mitochondrial genetics and maternal inheritance pattern.


Enzyme replacement therapy with agalsidase alfa in children with Fabry disease.
To assess the effects of enzyme replacement therapy (ERT) in children with Fabry disease. Safety and efficacy of ERT with agalsidase alfa, 0.2 mg/kg infused over 40 minutes every 2 weeks for 23 weeks, were studied in a multicentre open-label trial in nine boys and four girls. Median age at the start of the study was 11.0 years (range 3.5-18 years). Fifty-four adverse events were reported in 11 patients. No serious adverse events related to ERT were reported. Twelve of the 54 adverse events were considered possibly or probably related to ERT. Infusion reactions (8 mild, 3 moderate) occurred in four boys, in seven infusions. One boy developed IgG antibodies, although he continued to make good clinical progress. At the end of the study, two of the four boys and the one girl on regular pain medication at baseline had stopped taking analgesics. Brief Pain Inventory (BPI) scores decreased in most patients by week 12 and were sustained until the end of the study. This change was greater in the boys, who had higher (worse) BPI scores at baseline. Pain-related quality of life (QoL) scores also decreased during the study. Plasma globotriaosylceramide concentrations and urinary globotriaosylceramide:sphingomyelin ratios decreased after 12 and 23 weeks of therapy, particularly in the boys. Increases in sweat volume were recorded in three out of five of the boys and in one of two girls tested after 23 weeks of treatment. ERT with agalsidase alfa in children with Fabry disease is well tolerated and, in the short term, appears to decrease pain and to improve pain-related QoL.


Systematic isolation of the genus Alternaria in the atmosphere of the "Grande Sao Paulo" area (Brazil).
The genus Alternaria has been isolated from the atmosphere of several countries and its spores are considered as having a strong allergenic power. The description of the genus isolation with a relative frequency was noted in the following countries: Mexico, the United States of America, Canada, Great Britain, Spain, South Africa, Denmark and Argentina. Many Brazilian authors have pointed out the rare incidence of this genus excluding Faria (5) who recorded in the city of Belo Horizonte, State of Minas Gerais, a frequency of 21.1%. Several authors have failed to detect Alternaria sp. spores in the atmosphere of the city of Sao Paulo. As we were studying in our laboratory the airborne fungi of a region called Greater Sao Paulo, where the collections were made in some boroughs of the main city and its neighborhood, we began to make findings of the Alternaria sp. This gave us the opportunity of studying its incidence in that area during a period of one year. We chose 11 areas of this region where we made weekly plate expositions performing 429 trials in all. The technique of collection was the usual one with the exposition of the Petri dishes containing Sabouraud Agar distributed 72 hours before. The collection was made from September 1973 to August 1974. The exposition period was of 5 minutes duration in the open air or in a protected place when it rained. The exposition height was 1 meter from the ground. The identification was made through the colony macroscopic morphology and through the Riddell technique. The Alternaria sp. was isolated in the mentioned area through all the months of the year with its highest frequency in August (35.8%) and its lowest in December 3.7%). The average relative frequency obtained was of 17%. The highest isolation frequency was attained in Winter, the lowest in Spring; in Summer and Autumn, it remained at a medium level. In relation to the studied areas, the Alternaria sp. was isolated in all of them with a minimum frequency of 8.3% in Campos Eliseos and with a maximum frequency of 28.9% in Osasco. The authors would like to emphasize this unusual finding because in the region studied the Alternaria sp. was previously considered absent.


Giant prostatic hyperplasia and its causes.
We present a case of a 59-year-old male who was admitted to the emergency department with urinary retention, with a history of lower urinary tract symptoms, with the value of serum prostate specific antigen level of 100 ng/mL and an estimated prostate size of 800 mL, according to magnetic resonance imaging. A prostate biopsy showed benign prostatic hyperplasia. Transvesical prostatectomy was performed, following additional procedure of transurethral resection of the prostate. To the best of our knowledge, this is the fourth highest prostate volume reported in medical literature. In this paper, we examine the factors that may have influenced the development of giant prostate hyperplasia.


Long-Term Outcomes in Patients with Not-Retrieval Inferior Vena Cava Filter Under Anticoagulation.
Since permanent inferior vena cava (IVC) filters increase deep vein thrombosis (DVT), filter retrieval should be performed as possible. Despite the guideline recommendation, IVC filters are not always retrieved in clinical practice. To date, many patients with not-retrieval IVC filters have been prescribed anticoagulant therapy, but the long-term prognosis, including venous thromboembolism (VTE) and bleeding events, remains unknown. In this study, 195 patients who underwent IVC filter implantation between 2006 and 2017 at 3 institutions in Niigata City have been investigated about their deaths, VTE recurrence, and bleeding events. After peaking 2009, the number of IVC filter implantation gradually decreased. During observational period, there were 158 patients with not-retrieval IVC filters (the overall retrieval rate of 19.0%). The not-retrieval group included significantly older and more patients with cancer compared to the retrieval group. Anticoagulation therapy was continued in 88% of the not-retrieval group. During a mean follow-up of 5.0 years, 6 symptomatic DVT events associated with inadequate control of anticoagulation and 13 bleeding events were observed. A total of 52 patients died and only the presence of cancer was prognostic risk factor. Although long-term anticoagulation therapy may be associated with bleeding events, there were few recurrent VTE under optimal anticoagulation. It is anticipated that even if the IVC filter cannot be retrieved, appropriate anticoagulation is useful for prevention of DVT recurrence despite the risk of bleeding.


Photoassociation spectroscopy of ultracold Cs below the 6P1/2 limit.
We have performed high precision photoassociation spectroscopy of ultracold cesium gas. Using trap-loss fluorescence detection and controlling the background cesium pressure we were able to photoassociate atoms into excited states of ultracold molecules with large detunings up to 56 cm(-1) below the Cs(6S(1/2)) + Cs(6P(1/2)) atomic asymptote. Vibrational progressions are assigned to 0(g)(-), 0(u)(+), and 1(g) long-range states. By fitting the spectral data to the LeRoy-Bernstein expression, the effective coefficients of the leading long-range interactions and the vibrational quantum number at dissociation are obtained. In addition we have observed spectral perturbations between states of the same symmetry belonging to different asymptotes (6P(1/2) and 6P(3/2)). The perturbations are manifested through irregular vibrational level spacings and are especially pronounced in the 0(u)(+) symmetry. Many observed rotational levels indicate d- and higher partial wave contributions to the photoassociation cross section in the presence of trapping laser light, while spectral regions with only weak features suggest nodes in the lower state wave functions corresponding to the two ground state atoms asymptote.


Phylogeographic evidence for the inter- and intracontinental dissemination of avian influenza viruses via migration flyways.
Genetically related highly pathogenic avian influenza viruses (HPAIVs) of H5N6 subtype caused outbreaks simultaneously in East Asia and Europe-geographically distinct regions-during winter 2017-2018. This situation prompted us to consider whether the application of phylogeographic analysis to a particular gene segment of AIVs could provide clues for understanding how AIV had been disseminated across the continent. Here, the N6 NA genes of influenza viruses isolated across the world were subjected to phylogeographic analysis to illustrate the inter- and intracontinental dissemination of AIVs. Those isolated in East Asia during winter and in Mongolia/Siberia during summer were comingled within particular clades of the phylogeographic tree. For AIVs in one clade, their dissemination in eastern Eurasia extended from Yakutia, Russia, in the north to East Asia in the south. AIVs in western Asia, Europe, and Mongolia were also comingled within other clades, indicating that Mongolia/Siberia plays an important role in the dissemination of AIVs across the Eurasian continent. Mongolia/Siberia may therefore have played a role in the simultaneous outbreaks of H5N6 HPAIVs in Europe and East Asia during the winter of 2017-2018. In addition to the long-distance intracontinental disseminations described above, intercontinental disseminations of AIVs between Eurasia and Africa and between Eurasia and North America were also observed. Integrating these results and known migration flyways suggested that the migration of wild birds and the overlap of flyways, such as that observed in Mongolia/Siberia and along the Alaskan Peninsula, contributed to the long-distance intra- and intercontinental dissemination of AIVs. These findings highlight the importance of understanding the movement of migratory birds and the dynamics of AIVs in breeding areas-especially where several migration flyways overlap-in forecasting outbreaks caused by HPAIVs.


Identification of plasma RGS18 and PPBP mRNAs as potential biomarkers for gastric cancer using transcriptome arrays.
Coding and noncoding RNAs serve a crucial role in tumorigenesis. Circulating RNAs have been recognized as a novel category of biomarkers for a variety of physiological and pathological conditions. To identify plasma RNA biomarkers for gastric cancer (GC), a genome-wide transcriptome analysis using GeneChip® Human Transcriptome Array, which contains probe sets covering exons of ~67500 coding and noncoding transcripts of annotated genes, was performed to screen for the RNAs that exhibited differential expression in the plasma samples of patients with GC and controls. The expression levels of 6 candidate RNAs, including regulator of G-protein signaling 18 (RGS18), integral membrane protein 2B, pro-platelet basic protein (PPBP), nucleosome assembly protein1-like 1, n324674 and ENST00000442382 were assessed in the plasma samples of 81 patients with GC and 77 healthy participants using reverse transcription-quantitative polymerase chain reaction. Furthermore, the expression levels of RGS18 and PPBP mRNAs were indicated to be significantly differentially expressed (P<0.0001) in an independent panel of plasma samples of 36 patients with GC compared with 34 healthy participants. The potential association of RGS18 and PPBP mRNA expression levels with clinicopathological features was subsequently analyzed. Receiver operating characteristic analysis indicated that the combination of these 2 mRNAs with an area under curve <0.812 was an improved indicator for gastric cancer compared with respective individual levels. The results of the present study indicate that RGS18 and PPBP mRNA expression was significantly downregulated in the plasma of patients with GC, and the combination of these 2 mRNAs may be a useful diagnostic or prognostic marker for GC.


Preclinical evidence that use of TRAIL in Ewing's sarcoma and osteosarcoma therapy inhibits tumor growth, prevents osteolysis, and increases animal survival.
Osteosarcoma and Ewing's sarcoma are high-grade neoplasms typically arising in the bones of children and adolescents. Despite improvement in therapy, the five-year survival rate is only 20% for patients not responding to treatment or presenting with metastases. Among new therapeutic strategies, the efficacy of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), a member of the TNF superfamily with strong antitumoral activity and minimal toxicity to most normal cells and tissues, was investigated by complementary approaches both in vitro and in preclinical models. The sensitivity of osteosarcoma and Ewing's sarcoma cell lines to TRAIL was investigated in vitro by determining TRAIL receptor expression together with TRAIL effects on cell viability and apoptosis. Complementary preclinical studies were carried out in respective tumor models by inoculation of osteosarcoma or Ewing's sarcoma tumor cells in paraosseous location. In addition, a model of lung nodule dissemination was developed by i.v. injection of osteosarcoma cells. In vitro, both osteosarcoma and Ewing's sarcoma cells that express the TRAIL death receptors were highly sensitive to TRAIL-induced caspase-8-mediated apoptosis. TRAIL administered in vivo by nonviral gene therapy inhibited primary bone tumor incidence and growth by 87% and prevented tumor-induced osteolysis, leading to a significant 2-fold increase in animal survival 40 days after tumor induction. Furthermore, TRAIL inhibited tumor nodule dissemination in lungs and increased survival in an osteosarcoma model. These findings suggest that TRAIL is a promising candidate for the development of new therapeutic strategies in the most frequent malignant primary bone tumors.


6-mercaptopurine promotes energetic failure in proliferating T cells.
The anticancer drug 6-mercaptopurine (6-MP) inhibits de novo purine synthesis and acts as an antiproliferative agent by interfering with protein, DNA and RNA synthesis and promoting apoptosis. Metabolic reprogramming is crucial for tumor progression to foster cancer cells growth and proliferation, and is regulated by mechanistic target of rapamycin (mTOR) and AMP-activated protein kinase (AMPK) as well as the oncogenes Myc and hypoxia inducible factor 1α (HIF-1α). We hypothesized that 6-MP impacts metabolic remodeling through its action on nucleotide synthesis. The aim of our study is to provide a comprehensive characterization of the metabolic changes induced by 6-MP in leukemic T cells. Our results indicate that exposition to 6-MP rapidly reduces intracellular ATP concentration, leading to the activation of AMPK. In turn, mTOR, an AMPK target, was inhibited, and the expression of HIF-1α and Myc was reduced upon 6-MP incubation. As a consequence of these inhibitions, glucose and glutamine fluxes were strongly decreased. Notably, no difference was observed on glucose uptake upon exposition to 6-MP. In conclusion, our findings provide new insights into how 6-MP profoundly impacts cellular energetic metabolism by reducing ATP production and decreasing glycolytic and glutaminolytic fluxes, and how 6-MP modifies human leukemic T cells metabolism with potential antiproliferative effects.


Seven-state rotation-symmetric number-conserving cellular automaton that is not isomorphic to any septenary one.
We consider two-dimensional cellular automata with the von Neumann neighborhood that satisfy two properties of interest from a modeling viewpoint: rotation symmetry (i.e., the local rule is invariant under rotation of the neighborhood by 90^{∘}) and number conservation (i.e., the sum of all the cell states is conserved upon every update). It is known that if the number of states k is smaller than or equal to six, then each rotation-symmetric number-conserving cellular automaton is isomorphic to some k-ary one, i.e., one with state set {0,1,...,k-1}. In this paper, we exhibit an example of a seven-state rotation-symmetric number-conserving cellular automaton that is not isomorphic to any septenary one. This example strongly supports our plea that research into multistate cellular automata should not only focus on those that have {0,1,...,k-1} as a state set.


Design, synthesis of pyridine coupled pyrimidinone/pyrimidinthione as anti-MRSA agent: Validation by molecular docking and dynamics simulation.
Methicillin Resistant Staphylococcus aureus (MRSA) is a major cause of severe hospital and infections acquired by the population and related morbidity and mortality. In this unique situation, there is a need of dynamic strong drug candidates to control MRSA diseases. Thus, the present work focuses on the synthesis and characterization of pyrimidinones and pyrimidinthiones coupled pyridine derivatives as anti-MRSA agent. The synthesized compounds were characterized by different spectroscopic techniques and evaluated against MRSA strain. Among them, 4e and 4 g possessed better antibacterial activity with MIC values of 10 μg and 8 μg respectively. The key determinant of the wide range beta-lactam resistance in MRSA strains is the Penicillin-Binding Protein 2a (PBP2a) but the gene encodes PBP2a which has a low affinity towards β-lactam antibiotics. Because of this, the present investigation focused on the mechanism of PBP2a protein binding studies by in-silico studies. The synthesized compounds showed very good interactions with PBP2A compared with standard drug Vancomycin, among them compound 4 g showed better interaction with the binding score of -9.8 kcal/mol. Antibacterial activity was validated with molecular docking and molecular dynamic simulation. Simulation results revealed that protein-ligand interactions of 4 g compound stably sustained up to 20,000ps.Communicated by Ramaswamy H. Sarma.


Efficacy of gentamicin 0.3% solution of oral erosions healing in patients with severe generalized recessive dystrophic epidermolysis bullosa and its impact on the expression of type VII collagen.
Epidermolysis bullosa (EB) is a rare genetic skin disorder inherited either in autosomal recessive (AR) or autosomal dominant (AD) manner and characterized by blistering of the skin and mucous membranes. According to a subtype of EB, the oral manifestations and dental involvement vary in frequency and in severity. The most severe dental problems occur in patients with junctional epidermolysis bullosa (JEB) and severe generalized dystrophic epidermolysis bullosa (RDEB) and involve enamel erosion and development of blisters followed by painful oral wounds. Oral mucosa lesions decrease patients' quality of life and may contribute to difficulties in nutrition leading to cachexia. Assessment of efficacy of gentamicin 0.3% solution in the healing and preventing of oral erosions in patients with RDEB and evaluating its impact on the expression of type VII collagen. The study included four female patients with RDEB, aged 16-42 who show different mutations in the COL7A1 gene and were administered the mouth rinse two times daily with a solution of 0.3% gentamycin for 4 consecutive weeks. Prior to and at the end of the study, the samples from oral mucosa were collected to estimate the expression of type VII collagen by immunofluorescence test. The clinical improvement of oral wounds healing and reduced number of new blisters and mucous membrane soreness as well as partial re-expression of type VII collagen was observed in all studied patients. Topical gentamicin therapy of oral cavity in RDEB patients resulted in clinical improvement of mucosal lesions and re-expression of collagen type VII.


Proteomic Analysis of Matched Formalin-Fixed, Paraffin-Embedded Specimens in Patients with Advanced Serous Ovarian Carcinoma.
The biology of high grade serous ovarian carcinoma (HGSOC) is poorly understood. Little has been reported on intratumoral homogeneity or heterogeneity of primary HGSOC tumors and their metastases. We evaluated the global protein expression profiles of paired primary and metastatic HGSOC from formalin-fixed, paraffin-embedded (FFPE) tissue samples. After IRB approval, six patients with advanced HGSOC were identified with tumor in both ovaries at initial surgery. Laser capture microdissection (LCM) was used to extract tumor for protein digestion. Peptides were extracted and analyzed by reversed-phase liquid chromatography coupled to a linear ion trap mass spectrometer. Tandem mass spectra were searched against the UniProt human protein database. Differences in protein abundance between samples were assessed and analyzed by Ingenuity Pathway Analysis software. Immunohistochemistry (IHC) for select proteins from the original and an additional validation set of five patients was performed. Unsupervised clustering of the abundance profiles placed the paired specimens adjacent to each other. IHC H-score analysis of the validation set revealed a strong correlation between paired samples for all proteins. For the similarly expressed proteins, the estimated correlation coefficients in two of three experimental samples and all validation samples were statistically significant (p < 0.05). The estimated correlation coefficients in the experimental sample proteins classified as differentially expressed were not statistically significant. A global proteomic screen of primary HGSOC tumors and their metastatic lesions identifies tumoral homogeneity and heterogeneity and provides preliminary insight into these protein profiles and the cellular pathways they constitute.


Clinical evaluation in advanced practice nursing education: using standardized patients in Health Assessment.
Clinical education is critically important because competency in practice ultimately will determine the future of advanced practice nursing. Skills taught in Health Assessment, the first in a series of clinical courses, exposed students to tools that form the basis on which other competencies are built. The availability of standardized patients, people who participate in enacting a simulated but seemingly "real life" clinical encounter in a realistic clinical setting for the benefit of student learning and/or evaluation, made this instructional development project possible. The underlying assumption of this project was that clinical advanced practice nursing student education is enhanced by using an authentic clinical environment, known as a simulation center, with standardized patients and by using one or more evaluation techniques with multiple evaluators (i.e., peer, self, faculty, standardized patient). The student clinical experience was expected to improve and overall learning to increase by this method. This improvement was reflected at the end-of-course evaluations and in the quality of the final videotaped physical examination, which was superior to previous years. Student and faculty satisfaction with this teaching-learning process exceeded all expectations.


The consumers' views of genetic counseling of hemophilia.
Twenty-five Scottish and 22 Canadian patients with hemophilia, and 15 Scottish and 14 Canadian carriers of hemophilia participated in the study. They were interviewed with respect to their experience of and attitudes to genetic counseling, perceptions of the counselor's role, and satisfaction with existing care for families with hemophilia. Most patients and carriers favor genetic counseling as part of general counseling offered to them by the hemophilia center and think that such counseling should include dealing with such issues as schooling, employment, emotions, and psychological problems. Prenatal testing and termination of pregnancy is at present unacceptable to most of the participants, although they do not object to the use of these methods in cases of illnesses more severe than hemophilia.


[Fertility preservation in female oncology patients].
Four women were referred to the department ofGynaecology for fertility preservation. A 33-year-old nulliparous woman with breast cancer stage pT1cN0M0 underwent an IVF-ICSI cycle; five embryos were frozen. Pre-implantation genetic diagnosis (PGD) because of BRCA2 gene mutation carriage was not carried out and more recently follow-up oocyte donation options are being considered. A second, 32-year-old nulliparous woman with breast cancer stage pT2N1M0 underwent an IVF cycle; seven embryos were frozen. The third patient was a 14-year-old girl with osteosarcoma of the distal femur, who underwent a laparoscopic unilateral ovariectomy, one day after referral, and cortical tissue was frozen. The fourth patient was a 33-year-old nulliparous woman without partner, with non-Hodgkin lymphoma stage IIA. She underwent laparoscopic ovariectomy and cortical tissue was frozen. Infertility due to cancer treatment and fertility preservation options should be discussed early in treatment planning. Patients' expectations and fertility preservation limitations are important to consider. Fertility preservation options can be conducted in specialised hospitals under institutional review board approval. It still has an experimental status.


[Effect of Tripterygium Glycosides Tablets on reproductive toxicity in female rats with Ⅱ type collagen induced arthritis].
The aim of this paper was to observe the toxic effect of Tripterygium Glycosides Tablets( TG) on the reproductive system of Ⅱ type collagen induced arthritis( CIA) male rats,and to explore the toxic mechanism preliminarily. Fifty SD rats were randomly divided into normal control group( Con),model group( CIA),Tripterygium Glycosides Tablets clinical equivalent dose groups of 1,2,4 times( 9,18,36 mg·kg-1),10 rats in each group,and were given by gavage once a day for 42 days after the first immunization.The organ indexes of uterine and ovarian were calculated on days 21 and 42. Histopathological and morphological changes of uterine and ovarian were observed under optical microscope. The concentration of estradiol( E2),follicle-stimulating hormone( FSH),luteinizing hormone( LH),17α-hydroxylase( CYP17 A1) and cytochrome P450 19 A1( CYP19 A1) in serum were detected by ELISA. Immunohistochemistry was used to observe the expression of Bax and Bcl-2 related proteins in the apoptosis pathway of uterus and ovary. The results showed that compared with the Con group,CIA group could reduce the number of uterine glands( P<0.05),but no significant changes were observed in other groups. Compared with the CIA group,there were no significant changes in the coefficients of uterus and ovary in the Tripterygium Glycosides Tablets groups. The number of uterine glands,total follicles in the ovary,mature follicles and corpus luteum,the distribution of blood vessels and mitochondria had a certain inhibitory trend,and also slightly increased the number of atresia follicles,but the histopathological quantitative indicators were not statistically different. Except that 2 times clinical dose of Tripterygium Glycosides Tablets could significantly reduce the content of CYP19 A1( P<0. 05) after 42 d administration,there were no significant changes in serum estrogen E2,FSH,LH and estrogen synthesis key enzymes CYP17 A1 in each administration group. Medium and high doses of Tripterygium Glycosides Tablets could increase the expression of apoptotic protein Bax in uterine and ovarian tissues( P<0. 05,P<0. 01),and all the administration groups could inhibit the expression of apoptotic inhibiting protein Bcl-2( P <0. 05,P<0. 01,P<0.001),42 d was more obvious than 21 d. In conclusion,4 times and less than 4 times Tripterygium Glycosides Tablets did not cause obvious toxicity and histopathological changes in the reproductive organs of CIA rats,but it could reduce the level of serum estrogen synthesis key enzyme CYP19 A1 and affect the content of apoptosis-related proteins Bax and Bcl-2 in uterus and ovary tissues. The relevant mechanism needs further study.


Hepatic estrogen receptor in human liver disease.
Human liver contains estrogen receptors which render it sensitive to estrogen. Specific hormone binding to cytosol and nuclei from normal liver containing such receptors is of high affinity, low capacity, saturable, and specific for steroidal and nonsteroidal estrogens. Although estrogens alter metabolism and may produce disease, little data is available concerning estrogen receptor levels found in diseased liver. Herein we report estrogen receptor levels in human female liver containing diseases associated with oral contraceptives. Binding studies demonstrated cytosolic and nuclear estrogen receptors in human hepatic adenoma and focal nodular hyperplasia. Nuclear estrogen receptor levels in neoplastic tissue were greater than those in normal tissue. In addition, one hepatic adenoma resected from a patient taking tamoxifen contained no cytosolic estrogen receptor, and nuclear estrogen receptor levels were significantly lower than those found in normal tissue. These differences in binding capacity suggest a potential for greater hormone responsiveness in neoplastic liver tissue.


Screening and modular design for metabolic pathway optimization.
Biological conversion of substrate sugars to a variety of products is an increasingly popular option for chemical transformation due to its high specificity and because of significant interest in the use of renewable feedstocks. However, pathway optimization through metabolic engineering is often needed to make such molecules economically at a relevant scale. Employing effective methods to search and narrow the immense pathway parameter space is essential to meet performance metrics such as high titer, yield and productivity with efficiency. This review focuses on two practices that increase the likelihood of finding a more advantageous pathway solution: implementing a screen to identify high producers and utilizing modular pathway design to streamline engineering efforts. While screens seek to couple product titer with a high-throughput measurement output, modular design aims to rationally construct pathways to allow parallel optimization of various units. Both of these methodologies have proven widely successful in metabolic engineering, with combinations of them resulting in synergistic enhancements to pathway optimization. This review will particularly highlight their utility for microbially derived acid and alcohol products, which are of interest as fuels and value added products.


Streptochlorin isolated from Streptomyces sp. Induces apoptosis in human hepatocarcinoma cells through a reactive oxygen species-mediated mitochondrial pathway.
Streptochlorin is a small molecule isolated from marine Streptomyces sp. that is known to have antiangiogenic and anticancer properties. In this study, we examined the effects of this compound on reactive oxygen species (ROS) production and the association of these effects with apoptotic tumor cell death, using a human hepatocarcinoma Hep3B cell line. The results of this study demonstrated that streptochlorin mediates ROS production, and that this mediation is followed by a decrease in the mitochondrial membrane potential (MMP, m), activation of caspase-3, and downregulation of antiapoptotic Bcl-2 protein. The quenching of ROS generation by N-acetyl-L-cysteine administration, a scavenger of ROS, reversed the streptochlorin-induced apoptosis effects via inhibition of ROS production, MMP collapse, and the subsequent activation of caspase-3. These observations clearly indicate that ROS are involved in the early molecular events in the streptochlorin-induced apoptotic pathway. Taken together, our data imply that streptochlorin-induced ROS is a key mediator of MMP collapse, which leads to the caspase-3 activation, culminating in apoptosis.


Effect of storage time on clinical efficacy of single-donor platelet units.
Decrements in platelet function and recovery are known to accumulate during the 5-day storage period, and the clinical response to platelets transfused after several days' storage has been suggested by some researchers to be less than that seen with platelets stored for shorter periods. The clinical response to single-donor platelet (SDP) units (as measured by corrected count increments [CCIs] and intertransfusion intervals) was investigated in autologous bone marrow transplant patients. Twenty-seven consecutive autologous bone marrow transplant patients with a variety of hematologic and solid organ malignancies were evaluated for posttransfusion CCIs after 419 SDP transfusions of units stored for 1 to 5 days. Patients were not excluded from the study because of clinical condition, such as fever or sepsis. The mean 15-minute posttransfusion CCI for SDP units stored for only 1 day (11,006 +/- 5,157) was no different than that for units stored for 5 days (10,225 +/- 4,481; p > 0.05); 24-hour posttransfusion CCIs were also not different if the SDP unit had been stored for 1 day or 5 days (6229 +/- 4489 vs. 4786 +/- 2759; p > 0.05) or for any intermediate period. Nor were intertransfusion intervals affected by storage time. While platelets may exhibit a progressive lesion during the 5-day storage period, these changes do not result in a decreased clinical response.


Examination of the prevalence of stimulus overselectivity in children with ASD.
Many individuals with autism spectrum disorders (ASD) display stimulus overselectivity, wherein a subset of relevant components in a compound stimulus controls responding, which impairs discrimination learning. The original experimental research on stimulus overselectivity in ASD was conducted several decades ago; however, interventions for children with ASD now typically include programming to target conditional discriminations in ways that might minimize the prevalence of stimulus overselectivity. The present study assessed 42 children who had been diagnosed or educationally identified with ASD using a discrimination learning assessment. Of these 42 children, 19% displayed overselective responding, which is a lower percentage than that seen in early research. Possible explanations for this decreased percentage, implications for intervention, and future directions for research are discussed.


Ground state of the easy-axis rare-earth kagome langasite Pr3Ga5SiO14.
We report muon spin relaxation and {69,71}Ga nuclear quadrupolar resonance local-probe investigations of the kagome compound Pr3Ga5SiO14. Small quasistatic random internal fields develop below 40 K and persist down to our base temperature of 21 mK. They originate from hyperfine-enhanced 141Pr nuclear magnetism which requires a nonmagnetic Pr3+ crystal-field (CF) ground state. In addition, we observe a broad maximum of the relaxation rate at approximately 10 K which we attribute to the population of the first excited magnetic CF level. Our results yield a Van Vleck paramagnet picture, at variance with the formerly proposed spin-liquid ground state.


Correction: A generalized method for alignment of block copolymer films: solvent vapor annealing with soft shear.
Correction for 'A generalized method for alignment of block copolymer films: solvent vapor annealing with soft shear' by Zhe Qiang et al., Soft Matter, 2014, 10, 6068-6076, https://doi.org/10.1039/C4SM00875H.


Road Traffic Anomaly Detection via Collaborative Path Inference from GPS Snippets.
Road traffic anomaly denotes a road segment that is anomalous in terms of traffic flow of vehicles. Detecting road traffic anomalies from GPS (Global Position System) snippets data is becoming critical in urban computing since they often suggest underlying events. However, the noisy ands parse nature of GPS snippets data have ushered multiple problems, which have prompted the detection of road traffic anomalies to be very challenging. To address these issues, we propose a two-stage solution which consists of two components: a Collaborative Path Inference (CPI) model and a Road Anomaly Test (RAT) model. CPI model performs path inference incorporating both static and dynamic features into a Conditional Random Field (CRF). Dynamic context features are learned collaboratively from large GPS snippets via a tensor decomposition technique. Then RAT calculates the anomalous degree for each road segment from the inferred fine-grained trajectories in given time intervals. We evaluated our method using a large scale real world dataset, which includes one-month GPS location data from more than eight thousand taxi cabs in Beijing. The evaluation results show the advantages of our method beyond other baseline techniques.


Diverse secondary C(sp3)-H bond functionalization via site-selective trifluoroacetoxylation of aliphatic amines.
We describe a coinage-metal-catalyzed site-selective oxidation of secondary C(sp3)-H bonds for aliphatic amine substrates. Broad amine scope, good functional compatibility and late-stage diversification are demonstrated with this method. The steric demand of the β-substituents controlled diastereoselectivities under this catalytic system. The site selectivity favors secondary C(sp3)-H bonds over tertiary ones underscoring the unique synthetic potential of this method.


Initial assessment of patients infected with human immunodeficiency virus: the yield and cost of laboratory testing.
Our objective was to determine the yield and cost of standardized laboratory testing of HIV-infected patients entering medical care after testing positive for HIV. An HIV staging and referral clinic in a municipal public hospital was our site for a cross-sectional study, and 308 patients were evaluated in the clinic between February 1, 1990 and October 1, 1991. Patients underwent standardized laboratory testing, including hematologic studies, serum chemistries, infectious disease serologies, and chest radiographs. The percentage of abnormal results for each test was determined. Abnormal results were stratified as mild or severe. They were also examined with regard to whether injection drug users or other patient subgroups had higher percentages of abnormalities. Changes and Medicare reimbursements for the tests were also determined. There were substantial numbers of abnormalities in all types of laboratory testing. Only 3% of patients had normal CD4 lymphocyte counts; 85% had counts of < 500/mm3, and 35% were < 200/mm3. Forty-four percent of patients had at least one abnormal hematologic study; 8% were severe. Nearly 75% had abnormal liver function tests; 20% of these were severe abnormalities. Fifteen percent of patients were PPD-positive, and > 50% were anergic. Fourteen percent had a positive nonspecific test for syphilis, and 7% had a positive confirmatory test. Fourteen percent of patients had an abnormal chest radiograph.(ABSTRACT TRUNCATED AT 250 WORDS)


cis-Clerodane-type furanoditerpenoids from Tinospora crispa.
Phytochemical and cytotoxicity investigations on organic solvent extracts of the aerial parts of Tinospora crispa have led to the isolation of 15 cis-clerodane-type furanoditerpenoids. Of these, nine compounds (1-9) were found to be new. Spectroscopic assignments of a previously reported compound, borapetoside A (13), were revised on the basis of HMQC and HMBC correlations. No discernible activity was observed when compounds 10-13 were subjected to evaluation in cytotoxicity assays against human prostate cancer (PC-3) and the normal mouse fibroblast (3T3) cell lines.


Effects of chronic exposure to Formaldehyde on micronucleus rate of bone marrow cells in male mice.
Formaldehyde (FA) is the major volatile organic chemical (VOC) present in indoor air, and a constituent known to be associated with sick building syndrome. In the present study, mice were exposed to different concentrations of FA (0, 1, 10 mg/ m3) through static inhalation for 2 hours per day for 20 weeks. The polychromatic erythrocytes/ normochromatic erythrocyte (PCE/NCE) ratio and the micronucleus rates in bone marrow cells were detected. Data indicated that the PCE/NCE ratio in two FA exposure groups were statistically significant lower than the negative control group (P <0.05), and the micronucleus rate in two FA exposure groups were not significantly higher than the control group (P>0.05). These results suggest that chronic static inhalation of FA can reduce the ratio of PCE/NCE in the mice bone marrow, but the effects to the mice bone micronucleus rate are not sure.


A Bioengineering Approach to Myopia Control Tested in a Guinea Pig Model.
To investigate the biocompatibility of an injectable hydrogel and its ability to control myopia progression in guinea pigs. The study used a hydrogel synthesized from acrylated hyaluronic acid with a conjugated cell-binding peptide and enzymatically degradable crosslinker. Seven-day-old guinea pigs were first form deprived (FD) with diffusers for 1 week. One group was kept as an FD-only control; two groups received a sub-Tenon's capsule injection of either hydrogel or buffer (sham surgery) at the posterior pole of the eye. Form deprivation treatments were then continued for 3 additional weeks. Treatment effects were evaluated in terms of ocular axial length and refractive error. Safety was evaluated via intraocular pressure (IOP), visual acuity, flash electroretinograms (ERG), and histology. Both hydrogel and sham surgery groups showed significantly reduced axial elongation and myopia progression compared to the FD-only group. For axial lengths, net changes in interocular difference (treated minus control) were 0.04 ± 0.06, 0.02 ± 0.09, and 0.24 ± 0.08 mm for hydrogel, sham, and FD-only groups, respectively (P = 0.0006). Intraocular pressures, visual acuities, and ERGs of treated eyes were not significantly different from contralateral controls. Extensive cell migration into the implants was evident. Both surgery groups showed noticeable Tenon's capsule thickening. Sub-Tenon's capsule injections of both hydrogel and buffer inhibited myopia progression, with no adverse effects on ocular health. The latter unexpected effect warrants further investigation as a potential novel myopia control therapy. That the hydrogel implant supported significant cell infiltration offers further proof of its biocompatibility, with potential application as a tool for drug and cell delivery.


The narrow vesicourethral angle measured on postoperative cystography can predict urinary incontinence after robot-assisted laparoscopic radical prostatectomy.
Radical prostatectomy is associated with complications including urinary incontinence. A significant association between specific features of the vesicourethral anastomosis and urinary incontinence after radical prostatectomy has been demonstrated. The aim of this study was to identify the most useful predictor of postoperative urinary incontinence after robot-assisted laparoscopic radical prostatectomy (RALP) according to the features of the vesicourethral anastomosis as determined by postoperative cystography. The final study cohort consisted of 150 patients. Postoperative cystography was performed within 1 week after RALP. The ratio between the longitudinal and horizontal lengths (L/H) of the bladder, the position of the urethrovesical junction (UVJ) and the bladder neck angle as seen on the cystogram were evaluated. Postoperative continence status was evaluated by a 1 h pad test 1 day after catheter removal and by the use of safety pads, retrieved retrospectively from patient records. The association between these variables and urinary incontinence was then analyzed. All patients were followed for at least 1 year postoperatively. The continence rates on the 1 h pad test and 1 month and 1 year after RALP were 31.3%, 56% and 93.3%, respectively. In multivariate analyses, urinary incontinence was significantly associated with nerve sparing, L/H and the vesical angle as determined on the 1 h pad test, but only with the vesical angle at 1 month and 1 year postoperatively. A narrow vesical angle measured on cystography is a useful predictor of postoperative urinary incontinence after RALP.


Fatal severe coronary artery stenosis in Williams syndrome: decision making using late gadolinium enhancement cardiovascular MRI.
Williams syndrome is a well-recognised congenital disorder characterised by cardiovascular, connective tissue, and central nervous system abnormalities. Coronary artery abnormalities are seen in patients with supravalvar aortic stenosis, but end-stage ischaemic heart disease is rare. We report a case of end-stage ischaemic heart disease due to severe coronary arterial stenosis, highlighting how cardiovascular MRI contributed to the management.


Evaluation of Anthelmintic Resistance and Exhaust Air Dust PCR as a Diagnostic Tool in Mice Enzootically Infected with Aspiculuris tetraptera.
The entry of infectious agents in rodent colonies occurs despite robust sentinel monitoring programs, strict quarantine measures, and stringent biosecurity practices. In light of several outbreaks with Aspiculuris tetraptera in our facilities, we investigated the presence of anthelmintic resistance and the use of exhaust air dust (EAD) PCR for early detection of A. tetraptera infection. To determine anthelmintic resistance, C57BL/6, DBA/2, and NCr nude mice were experimentally inoculated with embryonated A. tetraptera ova harvested from enzootically infected mice, followed by treatment with 150 ppm fenbendazole in feed, 150 ppm fenbendazole plus 5 ppm piperazine in feed, or 2.1 mg/mL piperazine in water for 4 or 8 wk. Regardless of the mouse strain or treatment, no A. tetraptera were recovered at necropsy, indicating the lack of resistance in the worms to anthelmintic treatment. In addition, 10 of 12 DBA/2 positive-control mice cleared the A. tetraptera infection without treatment. To evaluate the feasibility of EAD PCR for A. tetraptera, 69 cages of breeder mice enzootically infected with A. tetraptera were housed on a Tecniplast IVC rack as a field study. On day 0, 56% to 58% of the cages on this rack tested positive for A. tetraptera by PCR and fecal centrifugation flotation (FCF). PCR from EAD swabs became positive for A. tetraptera DNA within 1 wk of placing the above cages on the rack. When these mice were treated with 150 ppm fenbendazole in feed, EAD PCR reverted to pinworm-negative after 1 mo of treatment and remained negative for an additional 8 wk. The ability of EAD PCR to detect few A. tetraptera positive mice was investigated by housing only 6 infected mice on another IVC rack as a field study. The EAD PCR from this rack was positive for A. tetraptera DNA within 1 wk of placing the positive mice on it. These findings demonstrate that fenbendazole is still an effective anthelmintic and that EAD PCR is a rapid, noninvasive assay that may be a useful diagnostic tool for antemortem detection of A. tetraptera infection, in conjunction with fecal PCR and FCF.


MiR-218 increases sensitivity to cisplatin in esophageal cancer cells via targeting survivin expression.
Increasing evidence showed that microRNAs (miRNAs) were implicated in the chemical resistance of human cancers. We intended to investigate the role of miR-218 in cisplatin sensitivity of esophageal cancer cells. Quantitative real-time polymerase chain reaction (qRT-PCR) was carried out to analyze miR-218 expression in human esophageal cancer cell line Eca9706 and a cisplatin-resistant subline (ECa9706-CisR cells). The effects of miR-218 transfection on ECa9706 and ECa9706-CisR cell viability, including cell viability and apoptosis rate were confirmed using MTT assay, or flow cytometry, respectively. qRT-PCR was used to validate survivin as a direct target gene of miR-218 in our system. We found that miR-218 was significantly decreased in ECa9706-CisR cells compared with parent Eca9706 cells. Overexpression of miR-218 by mimics transfection would enhance cisplatin sensitivity evaluated by cell viability inhibition and apoptosis promotion. We validated here survivin as a direct target of miR-218 in ECa9706 cells, which might contribute to the chemoresistance of esophageal cancer cells to cisplatin. In summary, our data suggest that miR-218 might represent as a promising sensitizer of cisplatin therapy in clinical esophageal cancer patients.


microRNA-622 acts as a tumor suppressor in hepatocellular carcinoma.
microRNAs (miRNAs) are important regulators of tumor development and progression. In this study, we aimed to explore the expression and role of miR-622 in hepatocellular carcinoma (HCC). We found that miR-622 was significantly downregulated in human HCC specimens compared to adjacent noncancerous liver tissues. miR-622 downregulation was significantly associated with aggressive parameters and poor prognosis in HCC. Enforced expression of miR-622 significantly decreased the proliferation and colony formation and induced apoptosis of HCC cells. In vivo studies demonstrated that miR-622 overexpression retarded the growth of HCC xenograft tumors. Bioinformatic analysis and luciferase reporter assays revealed that miR-622 directly targeted the 3'-untranslated region (UTR) of mitogen-activated protein 4 kinase 4 (MAP4K4) mRNA. Ectopic expression of miR-622 led to a significant reduction of MAP4K4 expression in HCC cells and xenograft tumors. Overexpression of MAP4K4 partially restored cell proliferation and colony formation and reversed the induction of apoptosis in miR-622-overexpressing HCC cells. Inhibition of JNK and NF-κB signaling phenocopied the anticancer effects of miR-622 on HCC cells. Taken together, miR-622 acts as a tumor suppressor in HCC and restoration of miR-622 may provide therapeutic benefits in the treatment of HCC.


Production and characterization of oil-in-water emulsions containing droplets stabilized by beta-lactoglobulin-pectin membranes.
Oil-in-water emulsions containing droplets stabilized by beta-lactoglobulin (beta-Lg)-pectin membranes were produced using a two-stage process. A primary emulsion containing small droplets (d(32) approximately 0.3 microm) was prepared by homogenizing 10 wt % corn oil with 90 wt % aqueous solution (1 wt % beta-Lg, 5 mM imidazole/acetate buffer, pH 3.0) using a high-pressure valve homogenizer. The primary emulsion was then diluted with pectin solutions to produce secondary emulsions with a range of pectin concentrations (5 wt % corn oil, 0.45 wt % beta-Lg, 5 mM imidazole/acetate buffer, 0-0.22 wt % pectin, pH 3.0). The electrical charge on the droplets in the secondary emulsions decreased from +33 +/- 3 to -19 +/- 1 mV as the pectin concentration was increased from 0 to 0.22 wt %, which indicated that pectin adsorbed to the droplet surfaces. The mean particle diameter of the secondary emulsions was small (d(32) < 1 microm) at relatively low pectin concentrations (<0.04 wt %), but increased dramatically at higher pectin concentrations (e.g., d(32) approximately 13 microm at 0.1 wt % pectin), which was attributed to charge neutralization and bridging flocculation effects. Emulsions with relatively small mean particle diameters (d(32) approximately 1.2 microm at 0.1 wt % pectin) could be produced by disrupting flocs formed in secondary emulsions containing highly negatively charged droplets, for example, by sonication, blending, or homogenization. The particles in these emulsions probably consisted of small flocs containing a number of protein-coated droplets bound together by pectin molecules. These emulsions had good stability to further particle aggregation up to relatively high ionic strengths (< or =500 mM NaCl) and low pH (pH 3). The interfacial engineering technology used in this study could lead to the creation of food emulsions with improved physicochemical properties or stability.


Nanoscale imaging of photocurrent enhancement by resonator array photovoltaic coatings.
Nanoscale surface patterning commonly used to increase absorption of solar cells can adversely impact the open-circuit voltage due to increased surface area and recombination. Here, we demonstrate absorptivity and photocurrent enhancement using silicon dioxide (SiO2) nanosphere arrays on a gallium arsenide (GaAs) solar cell that do not require direct surface patterning. Due to the combined effects of thin-film interference and whispering gallery-like resonances within nanosphere arrays, there is more than 20% enhancement in both absorptivity and photocurrent. To determine the effect of the resonance coupling between nanospheres, we perform a scanning photocurrent microscopy based on a near-field scanning optical microscopy measurement and find a substantial local photocurrent enhancement. The nanosphere-based antireflection coating (ARC), made by the Meyer rod rolling technique, is a scalable and a room-temperature process; and, can replace the conventional thin-film-based ARCs requiring expensive high-temperature vacuum deposition.


Bis(monoacylglycerol)phosphate as a non-invasive biomarker to monitor the onset and time-course of phospholipidosis with drug-induced toxicities.
Drug-induced phospholipidosis (PL) is a phospholipid storage disorder characterized by the accumulation of multi-lamellar bodies (myeloid bodies) in tissues. A major unanswered question is whether PL represents a benign adaptive response, symptom or early event in drug toxicity. The absence of a non-invasive biomarker to monitor tissue PL has made it difficult to determine the prevalence and implications of PL in the clinic. As a result, the interpretation of PL in risk assessment remains uncertain in preclinical and clinical drug development. This review describes the rationale for bis(monoacylglycerol)phosphate (BMP) as a biomarker of PL and explores the potential links between PL and the toxicities of drugs. The similarities between the hypothesized roles of BMP in PL and Niemann-Pick type C disease are discussed. The potential implications of PL for cellular function are described in the context of drug-induced QT prolongation, myopathy and renal toxicity. A specific species of BMP, di-docosahexaenoyl-BMP, should be investigated further as a non-invasive biomarker to monitor the onset and time course of PL and to better understand the functional consequences which could contribute to the toxicities of drugs.


O-GlcNAcylation regulates neurofilament-light assembly and function and is perturbed by Charcot-Marie-Tooth disease mutations.
The neurofilament (NF) cytoskeleton is critical for neuronal morphology and function. In particular, the neurofilament-light (NF-L) subunit is required for NF assembly in vivo and is mutated in subtypes of Charcot-Marie-Tooth (CMT) disease. NFs are highly dynamic, and the regulation of NF assembly state is incompletely understood. Here, we demonstrate that human NF-L is modified in a nutrient-sensitive manner by O-linked-β-N-acetylglucosamine (O-GlcNAc), a ubiquitous form of intracellular glycosylation. We identify five NF-L O-GlcNAc sites and show that they regulate NF assembly state. NF-L engages in O-GlcNAc-mediated protein-protein interactions with itself and with the NF component α-internexin, implying that O-GlcNAc may be a general regulator of NF architecture. We further show that NF-L O-GlcNAcylation is required for normal organelle trafficking in primary neurons. Finally, several CMT-causative NF-L mutants exhibit perturbed O-GlcNAc levels and resist the effects of O-GlcNAcylation on NF assembly state, suggesting a potential link between dysregulated O-GlcNAcylation and pathological NF aggregation. Our results demonstrate that site-specific glycosylation regulates NF-L assembly and function, and aberrant NF O-GlcNAcylation may contribute to CMT and other neurodegenerative disorders.


Closed, one-stop intelligent and accurate particle characterization based on micro-Raman spectroscopy and digital microfluidics.
Monoclonal antibodies are prone to form protein particles through aggregation, fragmentation, and oxidation under varying stress conditions during the manufacturing, shipping, and storage of parenteral drug products. According to pharmacopeia requirements, sub-visible particle levels need to be controlled throughout the shelf life of the product. Therefore, in addition to determining particle counts, it is crucial to accurately characterize particles in drug product to understand the stress condition of exposure and to implement appropriate mitigation actions for a specific formulation. In this study, we developed a new method for intelligent characterization of protein particles using micro-Raman spectroscopy on a digital microfluidic chip (DMF). Several microliters of protein particle solutions induced by stress degradation were loaded onto a DMF chip to generate multiple droplets for Raman spectroscopy testing. By training multiple machine learning classification models on the obtained Raman spectra of protein particles, eight types of protein particles were successfully characterized and predicted with high classification accuracy (93%-100%). The advantages of the novel particle characterization method proposed in this study include a closed system to prevent particle contamination, one-stop testing of morphological and chemical structure information, low sample volume consumption, reusable particle droplets, and simplified data analysis with high classification accuracy. It provides great potential to determine the probable root cause of the particle source or stress conditions by a single testing, so that an accurate particle control strategy can be developed and ultimately extend the product shelf-life.


SDHD immunohistochemistry: a new tool to validate SDHx mutations in pheochromocytoma/paraganglioma.
Pheochromocytomas (PCC) and paragangliomas (PGL) may be caused by a germline mutation in 12 different predisposing genes. We previously reported that immunohistochemistry is a useful approach to detect patients harboring SDHx mutations. SDHA immunostaining is negative in SDHA-mutated tumors only, while SDHB immunostaining is negative in samples mutated on all SDHx genes. In some cases of SDHD or SDHC-mutated tumors, a weak diffuse SDHB labeling has however been described. Here, we addressed whether the same procedure could be applicable to detect patients with germline SDHD mutations, by testing two new commercially available anti-SDHD antibodies. We performed a retrospective study on 170 PGL/PCC in which we investigated SDHD and SDHB expression by immunohistochemistry. SDHx-mutated PGL/PCC showed a completely negative SDHB staining (23/27) or a weak cytoplasmic background (4/27). Unexpectedly, we observed that SDHD immunohistochemistry was positive in SDHx-deficient tumors and negative in the other samples. Twenty-six of 27 SDHx tumors (including the four weakly stained for SDHB) were positive for SDHD. Among non-SDHx tumors, 138/143 were positive for SDHB and negative for SDHD. Five cases showed a negative immunostaining for SDHB, but were negative for SDHD. Our results demonstrate that a positive SDHD immunostaining predicts the presence of an SDHx gene mutation. Because SDHB negative immunostaining is sometimes difficult to interpret in the case of background, the addition of SDHD positive immunohistochemistry will be a very useful tool to predict or validate SDHx gene variants in PGL/PCC.


Role of Chirality in Drugs: An Overview.
Stereochemistry has occupied a great role in the manufacture and development of pharmaceuticals. Chiral properties play an important role in the determination of pharmacological actions of the drug. In recent years, there is a considerable interest in chiral separation to isolate and examine both enantiomers. This article provides an overview about the stereochemistry and its role in drugs, and also, offers approved isolation methods for enantiomeric pairs.


Measurement of body composition changes during hemodialysis by bioimpedance analysis.
Monitoring of fluid exchanges during hemodialysis is essential for the safety and efficacy of the procedure. Bioimpedance analysis has been used for the measurement of body compartments in many settings, but experience during hemodialysis is relatively scare. In a prospective study of 28 cases of chronic renal failure treated by long-term hemodialysis on an outpatient basis, body composition studies were done immediately before and after the dialysis session. Methods included body weight, serum albumin, hematocrit, hemoglobin, sodium and osmolality. These variables were compared with impedance (BIA) findings, namely lean body mass, body fat, total water, extra and intra-cellular water, third space, and exchangeable sodium/potassium ratio. Patients lost weight and improved hematocrit, hemoglobin, osmolality and serum sodium during the therapy, and BIA measurements were consistent with these changes. Lean body mass and body fat markedly changed along the dialysis session (respectively -24.5% and + 7.5%), and these results were out of proportion regarding known gains and losses in that period. It is concluded that BIA was useful for monitoring water fluxes during hemodialysis, but provided unreliable information about body fat and lean body mass.


Small hepatocellular carcinoma: intratumor ethanol treatment using new needle and guidance systems.
Intratumor injection of absolute ethanol to treat small hepatocellular carcinoma sometimes results in incomplete necrosis of the tumor. Causes of this include inhomogeneous distribution of the ethanol and difficulty in identifying the tumor after previous ethanol injections. To solve these problems, the authors designed a multiple-side-hole needle for ethanol injection and implanted one or more small steel coils into the tumor before treatment to serve as a landmark. Six patients thus treated all showed adequate necrosis on follow-up computed tomography, biopsy, and angiography studies; initially elevated serum alpha-fetoprotein levels present in five patients were decreased. A resected surgical specimen obtained in one patient showed extensive necrosis of the tumor as well as of the surrounding healthy liver; only a small locus of equivocally viable cancer cells remained in the tumor margin.