Transient potential gradients and impedance measures of tethered bilayer lipid membranes: pore-forming peptide insertion and the effect of electroporation.
In this work, we present experimental data, supported by a quantitative model, on the generation and effect of potential gradients across a tethered bilayer lipid membrane (tBLM) with, to the best of our knowledge, novel architecture. A challenge to generating potential gradients across tBLMs arises from the tethering coordination chemistry requiring an inert metal such as gold, resulting in any externally applied voltage source being capacitively coupled to the tBLM. This in turn causes any potential across the tBLM assembly to decay to zero in milliseconds to seconds, depending on the level of membrane conductance. Transient voltages applied to tBLMs by pulsed or ramped direct-current amperometry can, however, provide current-voltage (I/V) data that may be used to measure the voltage dependency of the membrane conductance. We show that potential gradients >~150 mV induce membrane defects that permit the insertion of pore-forming peptides. Further, we report here the novel (to our knowledge) use of real-time modeling of conventional low-voltage alternating-current impedance spectroscopy to identify whether the conduction arising from the insertion of a polypeptide is uniform or heterogeneous on scales of nanometers to micrometers across the membrane. The utility of this tBLM architecture and these techniques is demonstrated by characterizing the resulting conduction properties of the antimicrobial peptide PGLa.


Dual role of interleukin-6 in regulating insulin sensitivity in murine skeletal muscle.
Cytokines are elevated in various insulin-resistant states, including type 2 diabetes and obesity, although the contribution of interleukin-6 (IL-6) in the induction of these diseases is controversial. We analyzed the impact of IL-6 on insulin action in murine primary myocytes, skeletal muscle cell lines, and mice (wild type and protein-tyrosine phosphatase 1B [PTP1B] deficient). IL-6 per se increased glucose uptake by activating serine/threonine protein kinase 11 (LKB1)/AMP-activated protein kinase/protein kinase B substrate of 160 kDa (AS160) pathway. A dual effect on insulin action was observed when myotubes and mice were exposed to this cytokine: additive with short-term insulin (increased glucose uptake and systemic insulin sensitivity) but chronic exposure produced insulin resistance (impaired GLUT4 translocation to plasma membrane and defects in insulin signaling at the insulin receptor substrate 1 [IRS-1] level). Three mechanisms seem to operate in IL-6-induced insulin resistance: activation of c-Jun NH(2)-terminal kinase 1/2 (JNK1/2), accumulation of suppressor of cytokine signaling 3 (socs3) mRNA, and an increase in PTP1B activity. Accordingly, silencing JNK1/2 with either small interfering RNA or chemical inhibitors impaired phosphorylation of IRS-1 (Ser307), restored insulin signaling, and normalized insulin-induced glucose uptake in myotubes. When using a pharmacological approach, liver X receptor agonists overcome IL-6-induced insulin resistance by producing downregulation of socs3 and ptp1b gene expression. Finally, the lack of PTP1B confers protection against IL-6-induced insulin resistance in skeletal muscle in vitro and in vivo, in agreement with the protection against the IL-6 hyperglycemic effect observed on glucose and insulin tolerance tests in adult male mice. These findings indicate the important role of IL-6 in the pathogenesis of insulin resistance and further implicate PTP1B as a potential therapeutic target in the treatment of type 2 diabetes.


Endoplasmic reticulum localization of Sec12p is achieved by two mechanisms: Rer1p-dependent retrieval that requires the transmembrane domain and Rer1p-independent retention that involves the cytoplasmic domain.
Yeast Sec12p is a type II transmembrane protein in the ER, which is essential for the formation of transport vesicles. From biochemical and morphological lines of evidence, we have proposed that Sec12p is localized to the ER by two mechanisms: static retention in the ER and dynamic retrieval from the early Golgi compartment. We have also shown that Rer1p, a membrane protein in the Golgi, is required for correct localization of Sec12p. In the present study, we have performed a systematic analysis to determine the ER localization signals in Sec12p corresponding to these two mechanisms. Both the transmembrane domain (TMD) and the NH2-terminal cytoplasmic domain of Sec12p show the ability to localize the protein to the ER. The effect of the TMD is potent and sufficient by itself for the ER localization and is strongly dependent on Rer1p. On the other hand, the cytoplasmic domain shows a moderate ER-localization capability which is independent of Rer1p. The rate of mannosyl modification has been measured to distinguish between retention and retrieval. The cytoplasmic domain significantly delays the transport from the ER to the cis-Golgi. In contrast, the TMD shows only a subtle retardation in the transport from the ER to the cis-Golgi but strictly prevents the transport beyond there. From these observations, we conclude that the TMD mainly acts as the retrieval signal and the cytoplasmic domain contains the retention signal. This study not only supports the two-mechanisms hypothesis but also provides powerful tools to dissect the two.


Evidence for the best treatment for displaced intra-articular calcaneal fractures.
In conclusion, this clinical scenario has proved to be a clinic conundrum. Unfortunately the surgical team may focus on the fracture and may overlook significant patient characteristics that could help in guiding optimal patient care. Operative management is the treatment of choice as long as complications can be minimized. Nonoperative care can lead to successful results in select patients in a high percentage of cases. Percutaneous fixation and arthroscopic reduction are new are- as for scientific study.


Spontaneous shaker rat mutant - a new model for X-linked tremor/ataxia.
The shaker rat is an X-linked recessive spontaneous model of progressive Purkinje cell (PC) degeneration exhibiting a shaking ataxia and wide stance. Generation of Wistar Furth (WF)/Brown Norwegian (BN) F1 hybrids and genetic mapping of F2 sib-sib offspring using polymorphic markers narrowed the candidate gene region to 26 Mbp denoted by the last recombinant genetic marker DXRat21 at 133 Mbp to qter (the end of the long arm). In the WF background, the shaker mutation has complete penetrance, results in a stereotypic phenotype and there is a narrow window for age of disease onset; by contrast, the F2 hybrid phenotype was more varied, with a later age of onset and likely non-penetrance of the mutation. By deep RNA-sequencing, five variants were found in the candidate region; four were novel without known annotation. One of the variants caused an arginine (R) to cysteine (C) change at codon 35 of the ATPase, Ca(2+) transporting, plasma membrane 3 (Atp2b3) gene encoding PMCA3 that has high expression in the cerebellum. The variant was well supported by hundreds of overlapping reads, and was found in 100% of all affected replicas and 0% of the wild-type (WT) replicas. The mutation segregated with disease in all affected animals and the amino acid change was found in an evolutionarily conserved region of PMCA3. Despite strong genetic evidence for pathogenicity, in vitro analyses of PMCA3(R35C) function did not show any differences to WT PMCA3. Because Atp2b3 mutation leads to congenital ataxia in humans, the identified Atp2b3 missense change in the shaker rat presents a good candidate for the shaker rat phenotype based on genetic criteria, but cannot yet be considered a definite pathogenic variant owing to lack of functional changes.


[Malignant tumors of external ear. Review of cases in 1991-2001].
Carcinoma of the external auditory canal is a rare tumor. Owing to the rarity of this disease, the literature provides very few publications of series. There is not a widely accepted staging system. The lack of a standarized staging system impedes meaningful interpretation of the literature. It is very difficult to compare different treatment strategies and results. The study is a retrospective case review. Seven patients with external auditory canal malignancies were studied. We used the staging system proposed by Arriaga et al. for both the Squamous cell carcinoma and non-SCC. We describe the risk factors, the symptoms, the treatment and the outcome and prognosis for our patients.


IS2aR, a computational tool to transform voxelized reference phantoms into patient-specific whole-body virtual CTs for peripheral dose estimation.
The risk of radiogenic cancer induction due to radiotherapy depends on the dose received by the patient's organs. Knowing the position of all organs is needed to assess this dose in a personalized way. However, radiotherapy planning computed tomography (pCT) scans contain truncated patient anatomy, limiting personalized dose evaluation. To develop a simple and freely available computational tool that adapts an ICRP reference computational phantom to generate a patient-specific whole-body CT for peripheral dose computations. Various bone-segmentation methods were explored onto fifteen pCTs, and the one with the highest Sørensen-Dice coefficient was implemented. The reference phantom is registered to the pCT, obtaining a registration transform matrix, which is then applied to create the whole-body virtual CT. Additional validation involved a comparison of absorbed doses to organs delineated on both the pCT and the virtual CT. A dedicated graphical user interface was designed and implemented to house the developed functions for i) selecting a registration region on which automatic bone segmentation and rigid registration will occur, ii) displaying the results of these processes under selectable views, and iii) exporting the final patient-specific whole-body CT. This software was termed IS2aR. The tested whole-body virtual CT generated by IS2aR fulfilled our requirements. IS2aR is a user-friendly computational software to create a personalized whole-body CT containing the original structures in the reference phantom. The personalized dose deposited in peripheral organs can be estimated further to assess second cancer induction risk in epidemiological studies.


Primary care training and the evolving healthcare system.
With growing numbers of patient-centered medical homes and accountable care organizations, and the potential implementation of the Patient Protection and Affordable Care Act, the provision of primary care in the United States is expanding and changing. Therefore, there is an urgent need to create more primary-care physicians and to train physicians to practice in this environment. In this article, we review the impact that the changing US healthcare system has on trainees, strategies to recruit and retain medical students and residents into primary-care internal medicine, and the preparation of trainees to work in the changing healthcare system. Recruitment methods for medical students include early preclinical exposure to patients in the primary-care setting, enhanced longitudinal patient experiences in clinical clerkships, and primary-care tracks. Recruitment methods for residents include enhanced ambulatory-care training and primary-care programs. Financial-incentive programs such as loan forgiveness may encourage trainees to enter primary care. Retaining residents in primary-care careers may be encouraged via focused postgraduate fellowships or continuing medical education to prepare primary-care physicians as both teachers and practitioners in the changing environment. Finally, to prepare primary-care trainees to effectively and efficiently practice within the changing system, educators should consider shifting ambulatory training to community-based practices, encouraging resident participation in team-based care, providing interprofessional educational experiences, and involving trainees in quality-improvement initiatives. Medical educators in primary care must think innovatively and collaboratively to effectively recruit and train the future generation of primary-care physicians.


Performance of Classification Systems for Age-Related Macular Degeneration in the Rotterdam Study.
To compare frequently used classification systems for age-related macular degeneration (AMD) in their abilty to predict late AMD. In total, 9066 participants from the population-based Rotterdam Study were followed up for progression of AMD during a study period up to 30 years. AMD lesions were graded on color fundus photographs after confirmation on other image modalities and grouped at baseline according to six classification systems. Late AMD was defined as geographic atrophy or choroidal neovascularization. Incidence rate (IR) and cumulative incidence (CuI) of late AMD were calculated, and Kaplan-Meier plots and area under the operating characteristics curves (AUCs) were constructed. A total of 186 persons developed incident late AMD during a mean follow-up time of 8.7 years. The AREDS simplified scale showed the highest IR for late AMD at 104 cases/1000 py for ages <75 years. The Rotterdam classification showed the highest IR at 89 cases/1000 py >75 years. The 3-Continent harmonization classification provided the most stable progression. Drusen area >10% ETDRS grid (hazard ratio 30.05, 95% confidence interval [CI] 19.25-46.91) was most prognostic of progression. The highest AUC of late AMD (0.8372, 95% CI: 0.8070-0.8673) was achieved when all AMD features present at baseline were included. Highest turnover rates from intermediate to late AMD were provided by the AREDS simplified scale and the Rotterdam classification. The 3-Continent harmonization classification showed the most stable progression. All features, especially drusen area, contribute to late AMD prediction. Findings will help stakeholders select appropriate classification systems for screening, deep learning algorithms, or trials.


Pseudocataplexy: gelastic-atonic seizures.
Gelastic-atonic seizures are characterized by laughing and then loss of muscle tone. They are difficult to differentiate from laughter-induced cataplexy and may be indicative of Niemann-Pick disease Type C or variants. We studied an 11-year-old girl who had gelastic-atonic seizures that were triggered by laughing. The nature of her attacks was established by ictal EEG ambulatory recordings at home. There was no evidence of Niemann-Pick disease.


HDAC1-Mediated lncRNA Stimulatory Factor of Follicular Development to Inhibit the Apoptosis of Granulosa Cells and Regulate Sexual Maturity through miR-202-3p-COX1 Axis.
Abnormal sexual maturity exhibits significant detrimental effects on adult health outcomes, and previous studies have indicated that targeting histone acetylation might serve as a potential therapeutic approach to regulate sexual maturity. However, the mechanisms that account for it remain to be further elucidated. Using the mouse model, we showed that Trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, downregulated the protein level of Hdac1 in ovaries to promote the apoptosis of granulosa cells (GCs), and thus arrested follicular development and delayed sexual maturity. Using porcine GCs as a cell model, a novel sexual maturity-associated lncRNA, which was named as the stimulatory factor of follicular development (SFFD), transcribed from mitochondrion and mediated by HDAC1, was identified using RNA sequencing. Mechanistically, HDAC1 knockdown significantly reduced the H3K27ac level at the -953/-661 region of SFFD to epigenetically inhibit its transcription. SFFD knockdown released miR-202-3p to reduce the expression of cyclooxygenase 1 (COX1), an essential rate-limited enzyme involved in prostaglandin synthesis. This reduction inhibited the proliferation and secretion of 17β-estradiol (E2) while promoting the apoptosis of GCs. Consequently, follicular development was arrested and sexual maturity was delayed. Taken together, HDAC1 knockdown-mediated SFFD downregulation promoted the apoptosis of GCs through the miR-202-3p-COX1 axis and lead to delayed sexual maturity. Our findings reveal a novel regulatory network modulated by HDAC1, and HDAC1-mediated SFFD may be a promising new therapeutic target to treat delayed sexual maturity.


Regulation of complement protein biosynthesis in mononuclear phagocytes.
Proteins of the complement system (with the exception of the terminal components C6-9) are synthesized in mononuclear phagocytes. The extrahepatic macrophage is therefore an important local source of the complement proteins which may serve as a first-line host defence mechanism. Net synthesis and secretion of complement by these cells is a function of maturation of the mononuclear phagocytic series, the tissue from which the cells are isolated, and the state of macrophage activation. To define some of the mechanisms for regulation of complement gene expression in mononuclear phagocytes, the major histocompatibility complex class III genes and C3 have been investigated. These genes are expressed constitutively in hepatocytes and monocytes/macrophages. In the mononuclear phagocyte, interferon-gamma, at physiological concentrations, effects a dose- and time-dependent increase in factor B and C2 mRNA and a corresponding increase in factor B and C2 biosynthesis. This effect is specific inasmuch as the expression of other genes (e.g. C3) is decreased by interferon-gamma, and interferon-alpha and beta at concentrations one to two logs greater have only a minimal effect on C2 and factor B gene expression. Endotoxin acting directly on monocytes has qualitatively different effects on expression of the complement genes. These complex regulatory mechanisms are being investigated with the use of murine fibroblasts transfected with human DNA bearing the relevent complement genes.


Scanning Electron Microscopic Evaluation of Root Canal Surfaces Prepared with LightSpeed & Endowave Rotary System.
The purpose of this in vitro study was to evaluate and compare the cleaning efficiency on root canal surfaces prepared with LightSpeed and Endowave rotary instruments. A total of 30 freshly extracted human mandibular premolars were subjected for the study. They were divided into two groups, each group consisting of 15 teeth. Group 1: The canals were prepared with LightSpeed system. Group 2: Endowave rotary system. All the groups were prepared according to manufacturer's recommendation, using 5.25% sodium hypochlorite and 17% EDTA (dent wash, prime dent) alternately as an irrigants. Crowns of each tooth were removed with diamond disks at the level of cement enamel junction. Canal length was determined by placing a size 10 K-file. The working length was 0.5 mm short of canal length. Two longitudinal grooves were prepared on the lingual and buccal surfaces of each root to facilitate vertical splitting with a chisel after canal instrumentation. The sections were then observed under SEM for presence or absence of debris and smear layer and the photographs were taken at coronal, middle and apical 1/3 with a magnification of 200 X and 1000X respectively. The scores recorded were statistically analysed using one-way-ANOVA and Mann- Whitney test. There was statistically significant difference between two groups. All groups resulted in significantly more debris and smear layer in the apical 1/3 of the canal, compared to coronal and middle 1/3 (p, <0.01). The study demonstrated that, LightSpeed instrumentation removed debris and smear layer more effectively on root canal surface than compared to Endowave instrument.


Effects of recurrent middle ear effusion in preschool years on language and learning.
Eighteen healthy 8- to 11-year-old children with normal hearing and histories of recurrent middle ear effusion (MEE) before the age of 5 years were evaluated by a battery of audiologic, psychologic, language, and achievement tests. Each of these children was compared to a non-MEE sibling who was tested at the same age. None of the children had been diagnosed as learning disabled. Both groups tested in the bright normal range of verbal ability on the WISC-R. Paired comparison of sibling data revealed that the MEE subjects had deficits in verbal ability, auditory decoding, and spelling skills compared with control siblings. Significant strength in visual sequential memory in the MEE subjects suggests that the availability of compensatory strategies for auditory deficits in these subjects. Sex and/or birth order differences did not explain the differences observed between siblings.


Mutual Inhibition with Few Inhibitory Cells via Nonlinear Inhibitory Synaptic Interaction.
In computational neural network models, neurons are usually allowed to excite some and inhibit other neurons, depending on the weight of their synaptic connections. The traditional way to transform such networks into networks that obey Dale's law (i.e., a neuron can either excite or inhibit) is to accompany each excitatory neuron with an inhibitory one through which inhibitory signals are mediated. However, this requires an equal number of excitatory and inhibitory neurons, whereas a realistic number of inhibitory neurons is much smaller. In this letter, we propose a model of nonlinear interaction of inhibitory synapses on dendritic compartments of excitatory neurons that allows the excitatory neurons to mediate inhibitory signals through a subset of the inhibitory population. With this construction, the number of required inhibitory neurons can be reduced tremendously.


Adaptive, behavioral intervention impact on weight gain, physical activity, energy intake, and motivational determinants: results of a feasibility trial in pregnant women with overweight/obesity.
Interventions have modest impact on reducing excessive gestational weight gain (GWG) in pregnant women with overweight/obesity. This two-arm feasibility randomized control trial tested delivery of and compliance with an intervention using adapted dosages to regulate GWG, and examined pre-post change in GWG and secondary outcomes (physical activity: PA, energy intake: EI, theories of planned behavior/self-regulation constructs) compared to a usual care group. Pregnant women with overweight/obesity (N = 31) were randomized to a usual care control group or usual care + intervention group from 8 to 2 weeks gestation and completed the intervention through 36 weeks gestation. Intervention women received weekly evidence-based education/counseling (e.g., GWG, PA, EI) delivered by a registered dietitian in a 60-min face-to-face session. GWG was monitored weekly; women within weight goals continued with education while women exceeding goals received more intensive dosages (e.g., additional hands-on EI/PA sessions). All participants used mHealth tools to complete daily measures of weight (Wi-Fi scale) and PA (activity monitor), weekly evaluation of diet quality (MyFitnessPal app), and weekly/monthly online surveys of motivational determinants/self-regulation. Daily EI was estimated with a validated back-calculation method as a function of maternal weight, PA, and resting metabolic rate. Sixty-five percent of eligible women were randomized; study completion was 87%; 10% partially completed the study and drop-out was 3%. Compliance with using the mHealth tools for intensive data collection ranged from 77 to 97%; intervention women attended > 90% education/counseling sessions, and 68-93% dosage step-up sessions. The intervention group (6.9 kg) had 21% lower GWG than controls (8.8 kg) although this difference was not significant. Exploratory analyses also showed the intervention group had significantly lower EI kcals at post-intervention than controls. A theoretical, adaptive intervention with varied dosages to regulate GWG is feasible to deliver to pregnant women with overweight/obesity.


White matter abnormalities in bipolar disorder: insights from diffusion tensor imaging studies.
Diffusion tensor imaging (DTI) is a neuroimaging technique with the potential to elucidate white matter abnormalities. Recently, it has been applied to help in better understanding of the pathophysiology of bipolar disorder (BD). This review sought to synthesise existing literature on DTI studies in BD, summarise current findings and highlight brain regions that have consistently been implicated in BD, as well as posit possible future directions for DTI research in BD. The extant findings from this review suggest loss of white matter network connectivity as a possible phenomenon associated with bipolar disorder, involving prefrontal and frontal regions, projection, associative and commissural fibres, with sparse and less consistent evidence implicating the subcortical and non-frontal lobes of the brain. There are some differences in the direction of changes observed in white matter indices, and these may be attributed to factors including sample heterogeneity and limitations of DTI techniques. The possible roles of the parietal, temporal and occipital lobes and subcortical regions in BD await further investigation. Studies of bipolar disorder using DTI lag behind other neuropsychiatric diseases such as schizophrenia, but DTI research in BD is fast gaining pace. The emerging trends from these DTI findings underscore the importance of further research to unravel the underlying neural mechanisms and clinico-anatomical correlations involving white matter in BD.


Timing and Delivery of Fertility Preservation Information to Transgender Adolescents, Young Adults, and Their Parents.
This study aimed to examine transgender adolescents and young adults' (AYA) and their parents' preferences regarding fertility preservation (FP) information provision and discussion timing. Data were derived from two separate studies: an online survey and semistructured qualitative interviews. Survey data were analyzed using descriptive statistics and interview data using conventional content analysis. Survey participants (AYA: 88% and parents: 93%) preferred gender clinic physicians provide FP information, and nearly one-third endorsed mental health professionals (AYA: 28% and parents: 26%) or fertility specialists (AYA: 23% and parents: 30%). Interview participants' FP discussion timing preferences ranged from the initial clinic visit, follow-up visits, before medical intervention, to mentioning FP early but deferring in-depth discussion to follow-up visits. Gender clinic physicians, mental health professionals, and fertility specialists should be prepared to discuss FP with transgender AYA and their parents. Opinions varied regarding when to provide FP information; therefore, discussion timing may need to be individualized.


Transforming growth factor-beta (TGF-beta) and TGF-beta-associated kinase 1 are required for R-Ras-mediated transformation of mammary epithelial cells.
Transforming growth factor-beta (TGF-beta) cooperates with oncogenic members of the Ras superfamily to promote cellular transformation and tumor progression. Apart from the classic (H-, K-, and N-) Ras GTPases, only the R-Ras subfamily (R-Ras, R-Ras2/TC21, and R-Ras3/M-Ras) has significant oncogenic potential. In this study, we show that oncogenic R-Ras transformation of EpH4 cells requires TGF-beta signaling. When murine EpH4 cells were stably transfected with a constitutively active R-Ras(G38V) mutant, they were no longer sensitive to TGF-beta-mediated growth inhibition and showed increased proliferation and transformation in response to exogenous TGF-beta. R-Ras/EpH4 cells require TGF-beta signaling for transformation to occur and they produce significantly elevated levels of endogenous TGF-beta, which signals in an autocrine fashion. The effects of TGF-beta are independent of Smad2/3 activity and require activation of TGF-beta-associated kinase 1 (TAK1) and its downstream effectors c-Jun NH(2)-terminal kinase and p38 mitogen-activated protein kinase as well as the phosphoinositide 3-kinase/Akt and mammalian target of rapamycin pathways. Thus, TAK1 is a novel link between TGF-beta signaling and oncogenic R-Ras in the promotion of tumorigenesis.


Ultrasound and the Complete Urologist Part 2: Outcomes.
Outcome studies help establish the benefits of tests or procedures that can change the delivery of medical care. Through this discourse, the current status of outcomes with respect to the use of ultrasound imaging for AMH as well as its general use during a routine urologic physical examination are explored. The changes in today's health care environment focus on cost reduction and outcomes that produce value in the context of patient satisfaction. The question is whether we have to wait for outcome results (that can take a lengthy period of time to achieve) before the benefits produced by ultrasound can be applied. This report reviews the literature and the role ultrasound can play in today's urologic health care environment. Supportive evidence demonstrates the value that ultrasound use brings to the urologic examining table for AMH and to daily urologic practice to the benefit of both patient and physician.


The exposure assessment in current time study: implementation, feasibility, and acceptability of real-time data collection in a community cohort of illicit drug users.
Objective. We describe the study design and evaluate the implementation, feasibility, and acceptability of an ecological momentary assessment (EMA) study of illicit drug users. Design. Four sequential field trials targeting observation of 30 individuals followed for a four week period. Participants. Participants were recruited from an ongoing community-cohort of current or former injection drug users. Of 113 individuals enrolled, 109 completed study procedures during four trials conducted from November 2008 to May 2013. Methods. Hand-held electronic diaries used in the initial trials were transitioned to a smartphone platform for the final trial with identical data collection. Random-prompts delivered five times daily assessed participant location, activity, mood, and social context. Event-contingent data collection involved participant self-reports of illicit drug use and craving. Main Outcome Measures. Feasibility measures included participant retention, days of followup, random-prompt response rates, and device loss rate. Acceptability was evaluated from an end-of-trial questionnaire. Sociodemographic, behavioral, clinical, and trial characteristics were evaluated as correlates of weekly random-prompt response rates ≥80% using logistic regression with generalized estimating equations. Results. Study participants were a median of 48.5 years old, 90% African American, 52% male, and 59% HIV-infected with limited income and educational attainment. During a median followup of 28 days, 78% of 11,181 random-prompts delivered were answered (mean of 2.8 responses daily), while 2,798 participant-initiated events were reported (30% drug use events; 70% craving events). Self-reported acceptability to study procedures was uniformly favorable. Device loss was rare (only 1 lost device every 190 person-days of observation). Higher educational attainment was consistently associated with a higher response rate to random-prompts, while an association of HIV infection with lower response rates was not observed after accounting for differences in trial recruitment procedures. Conclusion. Near real-time EMA data collection in the field is feasible and acceptable among community-dwelling illicit drug users. These data provide the basis for future studies of EMA-informed interventions to prevent drug relapse and improve HIV treatment outcomes in this population.


F-18 FDG PET/CT findings of primary sinonasal hemangiopericytoma: rare location in a young adult patient.
Hemangiopericytoma is an unusual tumor that arises from vascular smooth muscle cells. It is mainly experienced in 4th to 5th decades of life and soft-tissue masses are frequently located in lower extremity, pelvis, and retroperitoneum. A 29-year-old man presented with a mass lesion in the right nasal cavity associated with shortness of breath as a result of nasal obstruction. An incisional biopsy was reported as sinonasal type hemangiopericytoma. The initial staging F-18 flourodeoxyglucose PET/CT successfully demonstrated severe hypermetabolism in the primary tumor location and mild-to-moderate hypermetabolism in the right upper jugular lymph node.


Process evaluation of a primary healthcare validation study of a culturally adapted depression screening tool for use by Aboriginal and Torres Strait Islander people: study protocol.
Process evaluations are conducted alongside research projects to identify the context, impact and consequences of research, determine whether it was conducted per protocol and to understand how, why and for whom an intervention is effective. We present a process evaluation protocol for the Getting it Right research project, which aims to determine validity of a culturally adapted depression screening tool for use by Aboriginal and Torres Strait Islander people. In this process evaluation, we aim to: (1) explore the context, impact and consequences of conducting Getting It Right, (2) explore primary healthcare staff and community representatives' experiences with the research project, (3) determine if it was conducted per protocol and (4) explore experiences with the depression screening tool, including perceptions about how it could be implemented into practice (if found to be valid). We also describe the partnerships established to conduct this process evaluation and how the national Values and Ethics: Guidelines for Ethical Conduct in Aboriginal and Torres Strait Islander Health Research is met. Realist and grounded theory approaches are used. Qualitative data include semistructured interviews with primary healthcare staff and community representatives involved with Getting it Right. Iterative data collection and analysis will inform a coding framework. Interviews will continue until saturation of themes is reached, or all participants are considered. Data will be triangulated against administrative data and patient feedback. An Aboriginal and Torres Strait Islander Advisory Group guides this research. Researchers will be blinded from validation data outcomes for as long as is feasible. The University of Sydney Human Research Ethics Committee, Aboriginal Health and Medical Research Council of New South Wales and six state ethics committees have approved this research. Findings will be submitted to academic journals and presented at conferences. ACTRN12614000705684.


Multimodal imaging probes based on Gd-DOTA conjugated quantum dot nanomicelles.
Recently, multimodal nanoparticles integrating dual- or tri-imaging modalities into a single hybrid nanosystem have attracted plenty of attention in biomedical research. Here, we report the fabrication of two types of multimodal micelle-encapsulated nanoparticles, which were systematically characterized and thoroughly evaluated in terms of their imaging potential and biocompatibility. Optical and magnetic resonance (MR) imaging probes were integrated by conjugating DOTA-gadolinium (Gd) derivative to quantum dot based nanomicelles. Two amphiphilic block copolymer micelles, amine-terminated mPEG-phospholipid and amine-modified Pluronic F127, were chosen as the capping agents because of their excellent biocompatibility and ability to prevent opsonization and prolong circulation time in vivo. Owing to their different hydrophobic-hydrophilic structure, the micellar aggregates exhibited different sizes and protection of core QDs. This work revealed the differences between these nanomicelles in terms of the stability over a wide range of pH, along with their cytotoxicity and the capacity for chelating gadolinium, thus providing a useful guideline for tailor-making multimodal nanoparticles for specific biomedical applications.


Effective state-based surveillance for multidrug-resistant organisms related to health care-associated infections.
In September 2008, the Council of State and Territorial Epidemiologists and the Centers for Disease Control and Prevention sponsored a meeting of public health and infection-control professionals to address the implementation of surveillance for multidrug-resistant organisms (MDROs)-particularly those related to health care-associated infections. The group discussed the role of health departments and defined goals for future surveillance activities. Participants identified the following main points: (1) surveillance should guide prevention and infection-control activities, (2) an MDRO surveillance system should be adaptable and not organism specific, (3) new systems should utilize and link existing systems, and (4) automated electronic laboratory reporting will be an important component of surveillance but will take time to develop. Current MDRO reporting mandates and surveillance methods vary across states and localities. Health departments that have not already done so should be proactive in determining what type of system, if any, will fit their needs.


Patient Reported Outcome Measure (PROM) of Quality of Life After Prostatectomy - Results from a 5-Year Study.
Prostate cancer is the most common cancer among men in Sweden, and treatment is negatively affecting the patients' quality of life. Even so, long term experiences are sparse and implications for nursing practice are little known. The aim of this study was to determine areas of functioning and factors impacting quality of life, QOL, during and five years after radical prostatectomy (RP) using a quality of life questionnaire and a specific module for prostate cancer. A longitudinal study was performed with consecutively included Swedish men from baseline and after RP treatment (n=222) from 2003 to 2011 to obtain their opinions on quality of life. Data was gathered through a mail out - mail in procedure at baseline, 3 months, 1-3 and 5 years after treatment with a response rate of 94.14% - 75.2%. One reminder was sent on each occasion. Identified areas with increased functioning after five years were emotional and social functioning. QOL ratings did not change over the years. Sexual activity and functioning decreased and hormonal treatment-related symptoms increased. Impact on QOL was found regarding emotional and social functioning, nausea/vomiting, pain and hormone-related symptoms. Increasing age, living with a partner and educational level had no significant impact on QOL. Implications for nursing are to initially focus on physical problems and at times for follow-up visits pay attention also to emotional and social aspects of life. To be able to make a difference in the patient's life, nurses need to bridge the gap between in-hospital treatment and everyday life outside hospital.


Impact of the legalisation of recreational cannabis use.
In recent years, there have been important legislative changes in many countries regarding the use of cannabis for medicinal and/or recreational purposes, which have facilitated access to it. Uruguay, Canada and some of the US states are the only jurisdictions that have legalised recreational consumption, applying different legislative models. The aim of this review is to analyse the effects that the legalisation of recreational cannabis has had on its use and its consequences. In general, the evidence accumulated to date indicates that the legalisation of cannabis has been associated with a decrease in the price of the substance, higher concentration of THC (potency), greater diversity of presentations for consumption, lower risk perception and an increase in consumption in adults and moderately in adolescents (even though it is illegal for them to consume), as well as an increase in the adverse consequences derived from cannabis consumption on public health. There has been a decrease in drug-related arrests, but the illegal market continues to be frequently used. No increase in the demand for treatment due to cannabis consumption has been detected. Therefore, these legislative changes have so far failed to achieve their main objectives, which were to suppress the illegal market and protect the most vulnerable groups, while on the contrary, they seem to imply an increase in some of the negative aspects associated with cannabis consumption. However, taking into account that most of these legislative changes have entered into force relatively recently, a longer follow-up period is required to be able to draw definitive conclusions.


An algorithm of calculating transport parameters of thermoelectric materials using single Kane band model with Riemann integral methods.
We develop an algorithm called SKBcal to conveniently calculate within minutes the thermoelectric transport parameters such as reduced Fermi level (η), electronic thermal conductivity (κe), lattice thermal conductivity (κl), Hall factor (A), effective mass (m*), quality factor (β) and theoretical zT within the framework of single Kane band (SKB) model. The generalized Fermi-Dirac integral for SKB model is integrated by left Riemann integral method. A concept of significant digits of relative error is involved to determine the accuracy of calculation. Furthermore, a combined program of "For" and "While" is coded to set up an iteration for refining the reduced Fermi level. To easily obtain the quality factor, we re-derive the expression into a formula related to carrier mobility. The results calculated by SKBcal are consistent with the data reported in the literatures.


Traumatic rupture of the thoracic aorta. A 20-year review: 1969-1989.
Acute traumatic injury of the thoracic aorta is a relatively common injury of deceleration accidents, usually high-speed motor vehicle accidents. The limiting factor to successful hospital management of these victims is early diagnosis and prompt treatment. Between the years 1969 and 1989, 116 patients (87 men and 29 women) were diagnosed with acute traumatic injury of the aorta. The mean age was 38.8 years, and the range was from 11 to 82 years. All patients who presented to the hospital emergency departments were included in the evaluation. The diagnosis of aortic rupture was firmly established by either aortography, surgery, or autopsy. The site of injury in 105 (90.5%) was the aortic isthmus, that is, distal to the origin of the left subclavian artery. In nine patients (7.7%) the injury was in the distal thoracic aorta. Two patients had injuries of the aortic arch. Associated injuries were present in 110 patients (95%); 16 patients (14%) had no external evidence of chest injury. Only 50 patients (43%) had the opportunity for definitive surgical management. The nonsurgical group comprised 66 patients (57%), and all of these died; the cause of death was unsuspected, suspected, or diagnosed after hospital admission from the emergency department. Of the surgical group, 36 patients (72%) survived, whereas 14 patients (28%) died. The surgical management comprised partial cardiopulmonary bypass in 31 patients that included seven deaths, simple aortic cross-clamping in 14 that included six deaths, and nonheparinized bypass in four that included one death. The incidence of paraplegia was 3.2% (one patient) with cardiopulmonary bypass, 28.5% (four patients) with simple aortic cross-clamping and none with nonheparinized bypass.(ABSTRACT TRUNCATED AT 250 WORDS)


Nuclei-specific thalamic connectivity predicts seizure frequency in drug-resistant medial temporal lobe epilepsy.
We assessed correlations between the resting state functional connectivity (RSFC) of different thalamic nuclei and seizure frequency in patients with drug-resistant medial temporal lobe epilepsy (mTLE). Seventeen patients with mTLE and 17 sex-/age-/handedness-matched controls participated. A seed-based correlation method for the resting-state FMRI data was implemented to get RSFC maps of 70 thalamic nuclei seed masks. Group statistics for individual RSFC for subjects and seed masks were performed to obtain within-group characteristics and between-group differences with age covariates. A linear regression was applied to test whether seizure frequency correlated with thalamic nuclear RSFC with the whole brain in mTLE patients. RSFC of thalamic nuclei showed spatially distinguishable connectivity patterns that reflected principal inputs and outputs that were derived from priori anatomical knowledge. We found group differences between normal control and mTLE groups in RSFC for nuclei seeds located in various subdivisions of thalamus. The RSFCs in some of those nuclei were strongly correlated with seizure frequency. Mediodorsal thalamic nuclei may play important roles in seizure activity or in the regulation of neuronal activity in the limbic system. The RSFC of motor- and sensory-relay nuclei may help elucidate sensory-motor deficits associated with chronic seizure activity. RSFC of the pulvinar nuclei of the thalamus could also be a key reflection of symptom-related functional deficits in mTLE.


Measuring the Advantage of Kalman-Weighted Averaging for Auditory Brainstem Response Hearing Evaluation in Infants.
The purposes of this study were to (a) measure the effects of Kalman-weighted averaging methods on auditory brainstem response (ABR) threshold, latency, and amplitude; (b) translate lab findings to the clinical setting; and (c) estimate cost savings when ABRs can be obtained in nonsedated infants. ABRs were recorded in 40 adults with normal hearing during induced motor noise conditions using the Kalman-weighted averaging method implemented on a commercial system, the Vivosonic Integrity (Vivosonic Inc., Toronto, Ontario, Canada). The device was then used to test 34 infants in awake and asleep states. The advantages of the Kalman-weighted averaging method were modeled in terms of time saved for conducting an ABR evaluation. Kalman-weighted ABR threshold estimates were 6-7 dB lower than with conventional methods during induced motor noise. When used to obtain ABRs in infants who were awake, the number of sweeps required to obtain a result was significantly greater than that required for a sleeping infant but well within the range for clinical application. The use of Kalman-weighted averaging provides a measurable advantage over conventional methods and may reduce costs for the pediatric audiology practice.


DNAPL flow and complex electrical resistivity evolution in saturated porous media: A coupled numerical simulation.
Induced Polarization (IP) is a non-intrusive geophysical method to monitor Dense Non-Aqueous Phase Liquid (DNAPL) contamination and remediation processes underground. In this study, an advanced numerical code simulating DNAPL flow and complex electrical resistivity is presented. The model was validated against existing IP results and image measurements that were carried out previously in a series of 2D tank experiment. Multiphase flow modeling in porous media is coupled with electrical current modeling to simulate the process of DNAPL migration and the associated IP response. This brings a broader view of the contamination in space and time compared to surface and borehole measurements, especially when the results are supported by field measurements or laboratory experiments. The simulations are developed in 3D and are performed in COMSOL Multiphysics®. The simulations using petrophysical relationships for in-phase and quadrature resistivity and the results of the experiments are in complete accordance with each other in the parts of the tank where the saturation of DNAPL is relatively low (i.e. especially in the cone of depression in the pumping scenario). However, the parts associated with high saturation of DNAPL show high errors between the in-phase resistivity simulations and the results from experiments. The present work can be regarded as a preliminary study toward further applications of coupled IP-multiphase flow for more accurate detection and monitoring of DNAPLs. It is suggested that the choice of tool/approach in this study be extended to larger-scale studies for further investigation.


The Thiamine diphosphate dependent Enzyme Engineering Database: a tool for the systematic analysis of sequence and structure relations.
Thiamine diphosphate (ThDP)-dependent enzymes form a vast and diverse class of proteins, catalyzing a wide variety of enzymatic reactions including the formation or cleavage of carbon-sulfur, carbon-oxygen, carbon-nitrogen, and especially carbon-carbon bonds. Although very diverse in sequence and domain organisation, they share two common protein domains, the pyrophosphate (PP) and the pyrimidine (PYR) domain. For the comprehensive and systematic comparison of protein sequences and structures the Thiamine diphosphate (ThDP)-dependent Enzyme Engineering Database (TEED) was established. The TEED http://www.teed.uni-stuttgart.de contains 12048 sequence entries which were assigned to 9443 different proteins and 379 structure entries. Proteins were assigned to 8 different superfamilies and 63 homologous protein families. For each family, the TEED offers multisequence alignments, phylogenetic trees, and family-specific HMM profiles. The conserved pyrophosphate (PP) and pyrimidine (PYR) domains have been annotated, which allows the analysis of sequence similarities for a broad variety of proteins. Human ThDP-dependent enzymes are known to be involved in many diseases. 20 different proteins and over 40 single nucleotide polymorphisms (SNPs) of human ThDP-dependent enzymes were identified in the TEED. The online accessible version of the TEED has been designed to serve as a navigation and analysis tool for the large and diverse family of ThDP-dependent enzymes.


Inhaled antibiotics in mechanically ventilated patients.
During the last decade, inhaled antibiotics, especially colistin, has been widely used worldwide as a therapeutic option, supplementary to conventional intravenous antibiotics, for the treatment of multidrug-resistant (MDR) Gram-negative nosocomial and ventilator-associated pneumonia (VAP). Antimicrobial aerosols are commonly used in mechanically ventilated patients with VAP, although information regarding their efficacy and optimal technique of administration has been limited. Recent studies showed that the administration of inhaled antibiotics in addition to systemic antibiotics provided encouraging results associated with low toxicity for the management of VAP mainly due to MDR Gram negative bacteria. Although the theory behind aerosolized administration of antibiotics seems to be sound, there are limited data available to support the routine use of this modality since very few randomized controlled trials (RCTs) have still examined the efficacy of this approach in patients with VAP. Additionally, this route of antibiotic delivery has not been approved until now neither by the FDA nor by the European Medicines Agency (EMEA) in patients with VAP. However, since the problem of VAP due to MDR bacteria has been increased worldwide RCTs are urgently needed in order to prove the safety, efficiency and efficacy of inhaled antimicrobial agents administered alone or in conjunction with parenteral antibiotics for the management of VAP in critically ill patients. Indeed, more data are needed to establish the appropriate role of inhaled antibiotics for the treatment of VAP.


Hedgehog regulated Slit expression determines commissure and glial cell position in the zebrafish forebrain.
Three major axon pathways cross the midline of the vertebrate forebrain early in embryonic development: the postoptic commissure (POC), the anterior commissure (AC) and the optic nerve. We show that a small population of Gfap+ astroglia spans the midline of the zebrafish forebrain in the position of, and prior to, commissural and retinal axon crossing. These glial ;bridges' form in regions devoid of the guidance molecules slit2 and slit3, although a subset of these glial cells express slit1a. We show that Hh signaling is required for commissure formation, glial bridge formation, and the restricted expression of the guidance molecules slit1a, slit2, slit3 and sema3d, but that Hh does not appear to play a direct role in commissural and retinal axon guidance. Reducing Slit2 and/or Slit3 function expanded the glial bridges and caused defasciculation of the POC, consistent with a ;channeling' role for these repellent molecules. By contrast, reducing Slit1a function led to reduced midline axon crossing, suggesting a distinct role for Slit1a in midline axon guidance. Blocking Slit2 and Slit3, but not Slit1a, function in the Hh pathway mutant yot (gli2DR) dramatically rescued POC axon crossing and glial bridge formation at the midline, indicating that expanded Slit2 and Slit3 repellent function is largely responsible for the lack of midline crossing in these mutants. This analysis shows that Hh signaling helps to pattern the expression of Slit guidance molecules that then help to regulate glial cell position and axon guidance across the midline of the forebrain.


[Morphological manifestations of tumor progression].
The paper reviews the theory of Foulds on the tumor progression and necessitates a wider application and study of this theory. The biological basis of the tumor progression consists in variability and heterogeneity of the tumor cells undergoing tissue selection resulting in the appearance of tumor cells with new properties. Morphological manifestations of the tumor progression subdivided into 4 groups are considered. Some aspects of application of the theory to histological diagnosis of tumors and its significance in treatment of cancer are described. It is emphasized that the theses of the tumor progression theory are essential for the development of basic research in oncology.


Tumor Growth Inhibition Modelling Based on Receptor Occupancy and Biomarker Activity of a New Bcl-2 Inhibitor in Mice.
The Bcl-2 inhibitor S 55746 ((S)-N-(4-hydroxyphenyl)-3-(6-(3-(morpholinomethyl)-1,2,3,4-tetrahydroisoquinoline-2-carbonyl)benzo[d][1,3]dioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide) is able to restore apoptosis functions impaired by tumorigenesis in mice. Data from pharmacokinetic (PK), biomarker, and tumor growth studies in a xenograft mouse model were considered for population modeling. The aim of the modeling exercise was to link the kinetics of the drug to the biomarker and tumor-size time profiles to better understand its dose-effect relationship. The PK, caspase kinetics, and tumor dynamics were successfully characterized by the proposed pharmacokinetic-pharmacodynamic model. The nonlinear plasma PK was best described by a two-compartment disposition model with both saturable absorption and elimination. Caspase was activated above the effective drug-concentration threshold (CTHRE ), at which near-maximal activity was reached. Increasing the dose did not increase the activation but better sustained it. Tumor growth followed a biphasic pattern, with caspase having an all-or-none inhibiting effect, consistent with the bistability property of the caspase pathway. For tumor eradication, the CTHRE in plasma was 2876 ng ml-1, and the relative caspase activity threshold (CaspTHRE) was 46.5. There was a strong relationship between the time spent above these thresholds and tumor growth inhibition. Tumor growth was inhibited by 50% when CaspTHRE was exceeded 13.8% of the time and when CTHRE was exceeded 8.1% of the time per dosing. This semimechanistic approach, based on experimental mice data and in vitro parameters, provides an interesting tool to quantify or simulate antitumor effects and, eventually, to plan phase 1 studies.


Generic pixel-wise speckle detection in Fourier-domain optical coherence tomography images.
We present a generic phase-domain processing method for detecting speckles in Fourier-domain optical coherence tomography (OCT) images. The physics behind the interferometry is revisited and analytically along with simulation results it indicates that the speckle formation comes with phase distortion to the complex OCT signal. The first and the second derivatives of phase along the imaging depth are then calculated for speckle identification. The phase-domain processing method was applied to images acquired by both spectral-domain OCT and swept-source OCT systems, and the experimental results show that this method enables pixel-wise speckle identification.


Dual mode microwave tool for dielectric analysis and thermal ablation treatment of organic tissue.
A dual mode tool design to analyze organic tissue and locally perform thermal ablation treatment is presented. The tool is made of an array of split-ring resonators. It can operate on a sensing mode to track the relative dielectric changes from the organic tissue and on a treatment mode to perform thermal ablation at different input powers. The measurements were done with phantoms of human tissue. The tool is able to focus a hot spot of approximately 0.2mm with a temperature of 109 °C at an input power of 10W.


Assay of Na-K-ATPase in cultured nephron epithelia.
Microenzymatic methods have been utilized in the past to quantify the activity of Na-K-ATPase in tubular segments of the mammalian nephron. The assay reported here measures the precipitated inorganic phosphate liberated by the hydrolysis of gamma-32P-ATP. Activity data in single nephron segments of the cortical collecting tubule (CCT) confirm previous work; specifically, first data on enzyme activity in small cultured cell populations derived from CCT demonstrate that the Na-carrier enzyme can be quantified in nephron cell cultures.


Challenges in the development of anti-epidermal growth factor receptor therapies in breast cancer.
Emerging results from clinical trials with epidermal growth factor receptor (EGFR) inhibitors indicate good tolerability and, at best, modest to no clinical activity in a variety of epithelial tumors, including breast carcinomas. Although the EGFR is widely expressed in epithelial cancers, there is no evidence of frequent EGFR alterations at the DNA level in human tumors. This lack of molecular evidence to suggest a pathogenic role for the EGFR in breast cancer questions the notion that use of single-agent EGFR inhibitors in this disease is a viable therapeutic approach. The lack of selection of EGFR-dependent breast tumors into trials with EGFR inhibitors suggests the possibility that these were not true-negative studies. A point of view regarding challenges in the development of anti-EGFR therapies for patients with breast cancer and possible approaches to overcome them is presented in this article.


Lingual tonsil abscess with parapharyngeal extension: a case report.
Lingual tonsil abscess is a rare disorder previously reported only once in the English literature. Because of their similar structure to that of the palatine tonsils, the lingual tonsils have the propensity to develop infection in the same way. The progression of infection, however, is different in that the lingual tonsils lack a capsule, thus preventing the formation of a peritonsillar abscess. Therefore, the only place for infection to spread is either into the tongue or into the parapharyngeal space. Here we present our experience with the latter, and we provide radiographic evidence of the disease. Lingual tonsil abscess, although rare, is an important potential cause of airway obstruction and must be considered in the case of a sore throat with a normal oropharyngeal exam.


Second generation fusion neutron time-of-flight spectrometer at optimized rate for fully digital data acquisition.
The progress on high-rate event recording of data is taken as starting point to revisit the design of fusion neutron spectrometers based on the TOF (time-of-flight) technique. The study performed was aimed at how such instruments for optimized rate (TOFOR) can be further developed to enhance the plasma diagnostic capabilities based on measurement of the 2.5 MeV dd neutron emission from D plasmas, especially the weak spectral components that depend on discrimination of extraneous events. This paper describes a design (TOFOR II) adapted for use with digital wave form recording of all detector pulses providing information on both amplitude (pulse height) and timing. The results of simulations are presented and the performance enhancement is assessed in comparison to the present.


Combating Obesity: Harnessing the Synergy of Postbiotics and Prebiotics for Enhanced Lipid Excretion and Microbiota Regulation.
Obesity is a chronic metabolic disease that can be induced by a high-fat diet (HFD) and predisposes to a variety of complications. In recent years, various bioactive substances, such as probiotics, prebiotics, and postbiotics, have been widely discussed because of their good anti-lipid and anti-inflammatory activities. In this paper, soybean protein isolate was used as a substrate to prepare the postbiotic. Compound prebiotics (galactose oligosaccharides, fructose oligosaccharides, and lactitol) preparation Aunulife Postbiotics and Prebiotics Composition (AYS) is the research object. Weight loss and bowel movements in mice induced by a high-fat diet were studied. Moreover, qualitative and quantitative analyses of small-molecule metabolites in AYS were performed to identify the functional molecules in AYS. After 12 weeks of feeding, the weight gain of mice that were fed with high-dose AYS (group H) and low-dose AYS (group L) from 4 to 12 weeks was 6.72 g and 5.25 g (p < 0.05), both of which were significantly lower than that of the high-fat diet (group DM, control group) group (7.73 g) (p < 0.05). Serum biochemical analysis showed that TC, TG, and LDL-C levels were significantly lower in mice from the H and L groups (p < 0.05). In addition, the fecal lipid content of mice in the L group reached 5.89%, which was significantly higher than that of the DM group at 4.02% (p < 0.05). The study showed that AYS changed the structure of the intestinal microbiota in mice on a high-fat diet, resulting in a decrease in the relative abundance of Firmicutes and Muribaculaceae and an increase in the relative abundance of Bacteroidetes, Verrucomicrobia, and Lactobacillus. The metabolomics study results of AYS showed that carboxylic acids and derivatives, and organonitrogen compounds accounted for 51.51% of the AYS metabolites, among which pantothenate, stachyose, betaine, and citrate had the effect of preventing obesity in mice. In conclusion, the administration of prebiotics and postbiotic-rich AYS reduces weight gain and increases fecal lipid defecation in obese mice, potentially by regulating the intestinal microbiota of mice on a high-fat diet.


Geographic disparities in prostate cancer outcomes--review of international patterns.
This study reviewed the published evidence as to how prostate cancer outcomes vary across geographical remoteness and area level disadvantage. A review of the literature published from January 1998 to January 2014 was undertaken: Medline and CINAHL databases were searched in February to May 2014. The search terms included terms of 'Prostate cancer' and 'prostatic neoplasms' coupled with 'rural health', 'urban health', 'geographic inequalities', 'spatial', 'socioeconomic', 'disadvantage', 'health literacy' or 'health service accessibility'. Outcome specific terms were 'incidence', 'mortality', 'prevalence', 'survival', 'disease progression', 'PSA testing' or 'PSA screening', 'treatment', 'treatment complications' and 'recurrence'. A further search through internet search engines was conducted to identify any additional relevant published reports. 91 papers were included in the review. While patterns were sometimes contrasting, the predominate patterns were for PSA testing to be more common in urban (5 studies out of 6) and affluent areas (2 of 2), higher prostate cancer incidence in urban (12 of 22) and affluent (18 of 20), greater risk of advanced stage prostate cancer in rural (7 of 11) and disadvantaged (8 of 9), higher survival in urban (8 of 13) and affluent (16 of 18), greater access or use of definitive treatment services in urban (6 of 9) and affluent (7 of 7), and higher prostate mortality in rural (10 of 20) and disadvantaged (8 of 16) areas. Future studies may need to utilise a mixed methods approach, in which the quantifiable attributes of the individuals living within areas are measured along with the characteristics of the areas themselves, but importantly include a qualitative examination of the lived experience of people within those areas. These studies should be conducted across a range of international countries using consistent measures and incorporate dialogue between clinicians, epidemiologists, policy advocates and disease control specialists.


Auditory Mismatch Negativity in Response to Changes of Counter-Balanced Interaural Time and Level Differences.
Interaural time differences (ITD) and interaural level differences (ILD) both signal horizontal sound source location. To achieve a unified percept of our acoustic environment, these two cues require integration. In the present study, we tested this integration of ITD and ILD with electroencephalography (EEG) by measuring the mismatch negativity (MMN). The MMN can arise in response to spatial changes and is at least partly generated in auditory cortex. In our study, we aimed at testing for an MMN in response to stimuli with counter-balanced ITD/ILD cues. To this end, we employed a roving oddball paradigm with alternating sound sequences in two types of blocks: (a) lateralized stimuli with congruently combined ITD/ILD cues and (b) midline stimuli created by counter-balanced, incongruently combined ITD/ILD cues. We observed a significant MMN peaking at about 112-128 ms after change onset for the congruent ITD/ILD cues, for both lower (0.5 kHz) and higher carrier frequency (4 kHz). More importantly, we also observed significant MMN peaking at about 129 ms for incongruently combined ITD/ILD cues, but this effect was only detectable in the lower frequency range (0.5 kHz). There were no significant differences of the MMN responses for the two types of cue combinations (congruent/incongruent). These results suggest that-at least in the lower frequency ranges (0.5 kHz)-ITD and ILD are processed independently at the level of the MMN in auditory cortex.


Apoplexy in sellar metastasis: a case report and review of literature.
Apoplexy in sellar metastasis is very rare with only a few case reports in literature. A case of apoplexy in sellar metastasis from follicular thyroid carcinoma is reported and the literature is briefly reviewed. The patient presented with sudden onset headache and bi-lateral loss of vision following thyroidectomy in a case of follicular carcinoma thyroid with proven sellar metastasis. CT scan showed hyperdense blood in sellar mass suggestive of apoplexy in sellar metastasis. The patient underwent early trans-sphenoidal decompression. Apoplexy in sellar metastasis, although very rare, can be clinico-radiologically indistinguishable from pituitary apoplexy and should be especially considered in the differential diagnosis of patients with known primary neoplastic disease. In view of similar patho-physiological mechanism, sellar metastasis with apoplexy should be managed in a similar manner as pituitary apoplexy.


Preoperative Function Assessment of Ex Vivo Kidneys with Supervised Machine Learning Based on Blood and Urine Markers Measured during Normothermic Machine Perfusion.
Establishing an objective quality assessment of an organ prior to transplantation can help prevent unnecessary discard of the organ and reduce the probability of functional failure. In this regard, normothermic machine perfusion (NMP) offers new possibilities for organ evaluation. However, to date, few studies have addressed the identification of markers and analytical tools to determine graft quality. In this study, function and injury markers were measured in blood and urine during NMP of 26 porcine kidneys and correlated with ex vivo inulin clearance behavior. Significant differentiation of kidneys according to their function could be achieved by oxygen consumption, oxygen delivery, renal blood flow, arterial pressure, intrarenal resistance, kidney temperature, relative urea concentration, and urine production. In addition, classifications were accomplished with supervised learning methods and histological analysis to predict renal function ex vivo. Classificators (support vector machines, k-nearest-neighbor, logistic regression and naive bayes) based on relevant markers in urine and blood achieved 75% and 83% accuracy in the validation and test set, respectively. A correlation between histological damage and function could not be detected. The measurement of blood and urine markers provides information of preoperative renal quality, which can used in future to establish an objective quality assessment.


Understanding the Mechanisms of Fe Deficiency in the Rhizosphere to Promote Plant Resilience.
One of the most significant constraints on agricultural productivity is the low availability of iron (Fe) in soil, which is directly related to biological, physical, and chemical activities in the rhizosphere. The rhizosphere has a high iron requirement due to plant absorption and microorganism density. Plant roots and microbes in the rhizosphere play a significant role in promoting plant iron (Fe) uptake, which impacts plant development and physiology by influencing nutritional, biochemical, and soil components. The concentration of iron accessible to these live organisms in most cultivated soil is quite low due to its solubility being limited by stable oxyhydroxide, hydroxide, and oxides. The dissolution and solubility rates of iron are also significantly affected by soil pH, microbial population, organic matter content, redox processes, and particle size of the soil. In Fe-limiting situations, plants and soil microbes have used active strategies such as acidification, chelation, and reduction, which have an important role to play in enhancing soil iron availability to plants. In response to iron deficiency, plant and soil organisms produce organic (carbohydrates, amino acids, organic acids, phytosiderophores, microbial siderophores, and phenolics) and inorganic (protons) chemicals in the rhizosphere to improve the solubility of poorly accessible Fe pools. The investigation of iron-mediated associations among plants and microorganisms influences plant development and health, providing a distinctive prospect to further our understanding of rhizosphere ecology and iron dynamics. This review clarifies current knowledge of the intricate dynamics of iron with the end goal of presenting an overview of the rhizosphere mechanisms that are involved in the uptake of iron by plants and microorganisms.


Molecular cloning, characterization, and complete nucleotide sequence of the gene for pneumolysin, the sulfhydryl-activated toxin of Streptococcus pneumoniae.
A recombinant lambda bacteriophage has been isolated that carries DNA from Streptococcus pneumoniae and expresses a potent hemolysin that has been shown to be pneumolysin, the sulfhydryl-activated toxin of the pneumococcus. Hemolytic activity is inhibited by cholesterol and neutralized by serum against streptolysin O. The cloned gene expresses two polypeptides (Mrs, 56,000 and 53,000) in an Escherichia coli in vitro transcription-translation system, and both are precipitated by the addition of anti-alveolysin serum and anti-streptolysin O serum in the presence of Staphylococcus aureus cells. Expression of pneumolysin occurs when the gene is cloned in both possible orientations in pUC8. The DNA sequence of a 5-kilobase ClaI fragment that carries the pneumolysin gene has been determined. An open reading frame was identified that encodes a polypeptide of 471 amino acids that is hydrophobic in character and has an N-terminal amino acid sequence which is identical to that deduced from amino acid sequencing of the purified protein. The predicted amino acid sequence of the polypeptide reveals a single cysteine residue located 44 residues from the C terminus. Putative promoter and ribosome binding sites have been identified 5' to the pneumolysin coding sequence.


Bitter taste inhibiting agents for whey protein hydrolysate and whey protein hydrolysate beverages.
Whey protein hydrolysates (WPH) are known for bioactivity and functionality, but WPH also have a distinct bitter taste. Identification of effective bitter taste inhibiting agents for WPH would broaden the use of this ingredient. The objective of this study was to evaluate the effectiveness of 24 documented bitter taste inhibitors for WPH. Two spray-dried WPH with different levels of hydrolysis (DH) were evaluated with each potential inhibitor. Quinine hydrochloride (quinine) was presented as a control with each WPH. Percent bitter taste inhibition was reported relative to quinine bitterness. Effective bitter taste inhibitors were subsequently evaluated in WPH beverages with vanilla and chocolate flavoring followed by descriptive analysis. The compounds evaluated did not inhibit bitter taste of quinine and the 2 WPH in a similar manner (P < 0.05). Effective bitter taste inhibitors (P < 0.05) of both WPH were sucralose, fructose, sucrose, adenosine 5' monophosphate (5'AMP), adenosine 5'monophosphate disodium (5'AMP Na(2) ), sodium acetate, monosodium glutamate, and sodium gluconate. Sodium chloride inhibited bitter taste of WPH with high DH but not WPH with low DH. Amino acids (l-Lysine, l-arginine) inhibited bitter taste of quinine but not WPH. All effective inhibitors in rehydrated WPH were also effective in the beverage applications. Sweeteners (fructose, sucralose, and sucrose) enhanced vanilla and chocolate flavors in beverages. Most salts and a nucleotide, while effective for bitter taste inhibition, suppressed vanilla and chocolate flavors and potentiated other flavors (that is, sour aromatic), and basic tastes (salty, sour). The bitter taste of whey protein hydrolysates (WPH) limits their use as ingredients. This study identified effective bitter taste inhibitors of WPH with different peptide composition and provides insights for effective bitter inhibitors for product applications with WPH.


MHC antigen expression in human oral squamous carcinoma cell lines.
This study quantified the constitutive and interferon-gamma (IFN-gamma) stimulated expression of MHC class I (HLA-ABC and beta 2 microglobulin) and class II antigens (HLA-DR, -DP, -DQ) on normal and malignant oral keratinocytes using radioimmunoassay and immunocytochemical techniques. Normal keratinocytes and three of four malignant cell lines (H103, H157, H314) expressed MHC class I antigens constitutively; IFN-gamma increased MHC class I expression with significant changes in normals, H157 and H314. Normal keratinocytes expressed significantly more constitutive MHC class I antigens than H103 and H157 and significantly more IFN-gamma stimulated MHC class I antigens than H103, H157 and H314. MHC class II antigens predominantly were not expressed constitutively on normals, H103 and H157 but, in H314, HLA-DR, -DP and -DQ antigens were demonstrated on 35, 11 and 5 per cent of cells, respectively, and resulted in a non-coordinated pattern of expression (HLA-DR greater than -DP = -DQ). IFN-gamma induced HLA-DR on normals, H103 and H157, whilst HLA-DP and -DQ remained undetectable. In H314, IFN-gamma enhanced HLA-DR, -DP and -DQ (significant increase of HLA-DQ) but the interrelationship between these antigens was maintained (HLA-DR greater than -DP = -DQ). Normal keratinocytes expressed significantly more IFN-gamma stimulated HLA-DR than H103 and H157 but significantly less HLA-DR than H314 under similar experimental conditions. One oral malignant cell line (H191) did not express MHC class I and MHC class II antigens either constitutively or in response to IFN-gamma. The results demonstrate aberrant patterns of MHC expression (absence, enhanced, diminished) in the different malignant oral keratinocyte cell lines.


Mapping and Characterization of HCMV-Specific Unconventional HLA-E-Restricted CD8 T Cell Populations and Associated NK and T Cell Responses Using HLA/Peptide Tetramers and Spectral Flow Cytometry.
HCMV drives complex and multiple cellular immune responses, which causes a persistent immune imprint in hosts. This study aimed to achieve both a quantitative determination of the frequency for various anti-HCMV immune cell subsets, including CD8 T, γδT, NK cells, and a qualitative analysis of their phenotype. To map the various anti-HCMV cellular responses, we used a combination of three HLApeptide tetramer complexes (HLA-EVMAPRTLIL, HLA-EVMAPRSLLL, and HLA-A2NLVPMVATV) and antibodies for 18 surface markers (CD3, CD4, CD8, CD16, CD19, CD45RA, CD56, CD57, CD158, NKG2A, NKG2C, CCR7, TCRγδ, TCRγδ2, CX3CR1, KLRG1, 2B4, and PD-1) in a 20-color spectral flow cytometry analysis. This immunostaining protocol was applied to PBMCs isolated from HCMV- and HCMV+ individuals. Our workflow allows the efficient determination of events featuring HCMV infection such as CD4/CD8 ratio, CD8 inflation and differentiation, HCMV peptide-specific HLA-EUL40 and HLA-A2pp65CD8 T cells, and expansion of γδT and NK subsets including δ2-γT and memory-like NKG2C+CD57+ NK cells. Each subset can be further characterized by the expression of 2B4, PD-1, KLRG1, CD45RA, CCR7, CD158, and NKG2A to achieve a fine-tuned mapping of HCMV immune responses. This assay should be useful for the analysis and monitoring of T-and NK cell responses to HCMV infection or vaccines.


Communicating science, mediating presence: reflections on the present, past and future of conferencing.
The move online of almost all meetings in 2020 in the wake of the COVID-19 pandemic threw into sharp relief the taken-for-granted centrality of conferences within scientific culture. While its impact on science has yet to be fully grasped, for the authors of this special issue, this situation held heuristic power for understanding the meanings and functions, now and historically, of international scientific conferencing. Ongoing discussions in the academic world about the pros and cons of virtual meetings bring out the central place of presence in these events and its mediation across space and time by modern infrastructures and technologies. From their rise in the mid-nineteenth century to the experiences of the present day, as well as in imagined futures, international conferences have been about communication. Following James Carey, they can be considered both as places for sharing knowledge and as rituals aimed at fostering and performing communities.


Serum visfatin as a metabolic biomarker in obese patients with gestational diabetes mellitus.
Visfatin is an adipokine produced and secreted by the adipose tissue. It exerts an insulin-like effect by the insulin receptor-1 and has a hypoglycemic effect. We aimed to investigate how serum visfatin changes in women with gestational diabetes mellitus (GDM), and whether it is predictive of neonatal outcomes. Visfatin levels were prospectively measured in peripheral blood serum by enzyme immunoassay in 210 pregnant women, 156 of which were diagnosed with GDM, 18 of which suffered from pregnancy-induced hypertension (PIH) and 36 healthy controls. Patients with obesity class II (median=2.562 ng/mL) and class III (median=6.2940 ng/mL) had higher serum visfatin than overweight patients (median=0.735 ng/mL); (Mann-Whitney U test, P=0.037 and P=0.023, respectively). In GDM patients with BMI above 30, serum visfatin was associated to glycosylated hemoglobin (Spearman correlation test, R=0.26, P=0.045). Women with BMI above 25 treated with insulin had lower serum visfatin levels than those treated with diet only (Mann-Whitney U test, P=0.045). No correlation was found between visfatin and parameters of lipid profile such as HDL, LDL, or triglycerides (Spearman correlation tests, R=-0.051, -0.1, 0.0019; P=0.54, 0.29, 0.98, respectively). We observed that visfatin was not associated with birth weight (Spearman correlation test, R=-0.014, P=0.86) or adverse neonatal outcome as measured by umbilical artery pH below 7.25 (Mann-Whitney U test, P=0.55) or Apgar score below 10 (Mann-Whitney U test, P=0.21). In GDM patients with higher BMI, serum visfatin was elevated, correlated positively with glycosylated hemoglobin, and decreased upon treatment with insulin therapy.


[Interaction of various tetrahydroisoquinoline alkaloids with opiate receptors in the rat hypothalamus and midbrain].
Using the method of competitive displacement of 3H-labelled naloxone, (D-ala2)-met enkephalinamide or (D-ala2)-D-leu-enkephalin by salsolinol, 1-methyl-6-hydroxy-1, 2, 3, 4-tetrahydro-beta-carboline or 1-methyl-6-methoxy-1, 2, 3, 4-tetrahydro-beta-carboline from opiate receptors, it was shown that the alkaloids studied were capable to cause specific interactions with rat hypothalamus and midbrain however, exhibiting distinctly less affinity as compared with morphine or its analogs. The sodium ratios, determined from the effective doses of the tetrahydroisoquinoline alkaloids corresponding to the alkaloid concentrations, which induce 50% displacement of 3H-naloxone from the opiate receptors in the presence or absence of 100 mM NaCl, have been found to be 0.75 for salsolinol and 3.6 for beta-carbolines studied. The data obtained suggest that salsolinol, similar to naloxone, is a "pure" morphine antagonist, whereas the beta-carbolines studied may be classified with the agonist-antagonist type. A considerable decrease in the affinity of mu-type opiate receptors has been found in presence of salsolinol in the incubation medium. The possible mechanisms of pharmacological action of the alkaloids and their relation to development of alcohol dependence and tolerance are discussed.


Differential subcellular localization of human MutY homolog (hMYH) and the functional activity of adenine:8-oxoguanine DNA glycosylase.
The post-replicative adenine:8-oxo-7,8-dihydroguanine (GO) mismatch is crucial for G:C to T:A transversion. This mismatch is corrected by Escherichia coli MutY which excises the adenine from A:GO. A candidate gene coding for the human counterpart of MutY has been cloned as hMYH. However, the function and enzyme activities of the gene product have not been identified. We previously demonstrated that an epitope-tagged hMYH protein behaves as a mitochondrial protein. In the present study, we have identified an alternative hMYH transcript, termed type 2, which differs in the exon 1 sequence of the known transcript (type 1). A nuclear localization for the type 2 protein was revealed by detection of epitope-tagged protein in COS-7 cells. Expression of both type 1 and type 2 transcripts was reduced in post-mitotic tissues. hMYH cDNA suppressed the mutator phenotype of E.coli mutY. In vitro expressed hMYH showed adenine DNA glycosylase activity toward the A:GO substrate. The protein can bind to A:GO, and to T:GO and G:GO without apparent catalysis. These results represent the first demonstration of the function of the hMYH gene product which is differentially transported into the nucleus or the mitochondria by alternative splicing


Epidemiological profile and genetic resistance of Neisseria gonorrhoeae infection in women in a poor region of São Paulo, Brazil.
Gonorrhea is a bacterial infection in the urogenital tract, transmitted by sexual or perinatal contact, caused by Neisseria gonorrhoeae, a gram-negative diplococcus. The present study evaluates the frequency of N. gonorrhoeae in women treated at Hospital Wladimir Arruda in poor area of São Paulo and also verifies the presence of genetic resistance against three antimicrobials of different classes: Tetracycline, Azithromycin and Ciprofloxacin. This is an observational and descriptive study with a quantitative approach. Samples were collected at Hospital Escola Wladimir Arruda. The volunteers are women from 16 to 65 years of age. Sociodemographic, gynecological, sexual and health data are collected through a questionnaire, their symptoms/clinical manifestation were requested by the medical records, and then the participant is referred for collection of samples of cervical vaginal smear. The samples were screened for N. gonorrhoeae (dcmH gene) and tested for resistance genes to Tetracycline, Azithromycin and Ciprofloxacin through PCR. In the total of 127 samples analyzed by Real-Time PCR, 23 were positive and correspond to a general prevalence of a gonococcal infection in the studied population of 17% (CI:95%), and the participants were married (43.4%), had active sexual life (56.5%) and did not use any type of condom during sexual intercourse (52.1%). The resistance to the tetM ribosomal gene was found in 14 samples, prevalence of 60% (CI= 95%). We have described a concerning frequency of N. gonorrhoeae infection in females attended in an outcare patient. Also, most of the strains detected presented resistance to one or more antimicrobials.


Caloric Restriction and the Nutrient-Sensing PGC-1α in Mitochondrial Homeostasis: New Perspectives in Neurodegeneration.
Mitochondrial activity progressively declines during ageing and in many neurodegenerative diseases. Caloric restriction (CR) has been suggested as a dietary intervention that is able to postpone the detrimental aspects of aging as it ameliorates mitochondrial performance. This effect is partially due to increased mitochondrial biogenesis. The nutrient-sensing PGC-1α is a transcriptional coactivator that promotes the expression of mitochondrial genes and is induced by CR. It is believed that many of the mitochondrial and metabolic benefits of CR are due to increased PGC-1α activity. The increase of PGC-1α is also positively linked to neuroprotection and its decrement has been involved in the pathogenesis of many neurodegenerative diseases. This paper aims to summarize the current knowledge about the role of PGC-1α in neuronal homeostasis and the beneficial effects of CR on mitochondrial biogenesis and function. We also discuss how PGC-1α-governed pathways could be used as target for nutritional intervention to prevent neurodegeneration.


Oesophageal tone in patients with achalasia.
The diagnosis and classification of oesophageal motility disorders is currently based on assessment of the phasic contractile activity of the oesophagus. Tonic muscular contraction of the oesophageal body (oesophageal tone) has not been well characterised. To quantify oesophageal tonic activity in healthy subjects and in patients with achalasia. Oesophageal tone was measured in 14 patients with untreated achalasia and in 14 healthy subjects. In eight patients with achalasia, oesophageal tone was again measured one month after either endoscopic or surgical treatment. Tonic wall activity was quantified by means of a flaccid intraoesophageal bag, 5 cm long and of 120 ml maximal capacity, which was placed and maintained 5 cm above the lower oesophageal sphincter and connected to an external electronic barostat. The experimental design included measurement of oesophageal basal tone and compliance as well as the oesophageal tone response to a nitric oxide donor (0.5 ml amyl nitrite inhalation). Oesophageal basal tone, expressed as the intrabag (intraoesophageal) volume at a minimal distending pressure (2 mm Hg), did not differ significantly between patients with achalasia and healthy controls (6.6 (2.5) ml versus 4.1 (0.8) ml, respectively). Oesophageal compliance (volume/pressure relation during intraoesophageal distension) was significantly increased in achalasia (oesophageal extension ratio: 3.2 (0.4) ml/mm Hg versus 1.9 (0.2) ml/mm Hg; p < 0.01). Amyl nitrite inhalation induced oesophageal relaxation both in patients and in controls, but the magnitude of relaxation was greater in the latter (intrabag volume increase: 15.3 (2.4) ml versus 36.2 (7.1) ml; p < 0.01). In patients with achalasia, oesophageal tonic activity, and not only phasic activity, is impaired. Although oesophageal compliance is increased, residual oesophageal tone is maintained so that a significant relaxant response may occur after pharmacological stimulation.


Cellular origin and microRNA profiles of circulating extracellular vesicles in different stages of diabetic nephropathy.
Diabetic nephropathy (DN) is a major complication of diabetes and the main cause of end-stage renal disease. Extracellular vesicles (EVs) are small cell-derived vesicles that can alter disease progression by microRNA (miRNA) transfer. In this study, we aimed to characterize the cellular origin and miRNA content of EVs in plasma samples of type 2 diabetes patients at various stages of DN. Type 2 diabetes patients were classified in three groups: normoalbuminuria, microalbuminuria and macroalbuminuria. The concentration and cellular origin of plasma EVs were measured by flow cytometry. A total of 752 EV miRNAs were profiled in 18 subjects and differentially expressed miRNAs were validated. Diabetic patients with microalbuminuria and/or macroalbuminuria showed elevated concentrations of total EVs and EVs from endothelial cells, platelets, leucocytes and erythrocytes compared with diabetic controls. miR-99a-5p was upregulated in macroalbuminuric patients compared with normoalbuminuric and microalbuminuric patients. Transfection of miR-99a-5p in cultured human podocytes downregulated mammalian target of rapamycin (mTOR) protein expression and downregulated the podocyte injury marker vimentin. Type 2 diabetes patients with microalbuminuria and macroalbuminuria display differential EV profiles. miR-99a-5p expression is elevated in EVs from macroalbuminuria and mTOR is its validated mRNA target.


Auto-detection of necessity for MRI-guided online adaptive replanning using a machine learning classifier.
MRI-guided adaptive radiation therapy (MRgART), particularly daily online adaptive replanning (OLAR) can substantially improve radiation therapy delivery, however, it can be labor-intensive and time-consuming. Currently, the decision to perform OLAR for a treatment fraction is determined subjectively. In this work, we develop a machine learning algorithm based on structural similarity index measure (SSIM) and change in entropy to quickly and objectively determine whether OLAR is necessary for a daily MRI set. A total of 109 daily MRI sets acquired on a 1.5T MR-Linac during MRgART for 22 pancreatic cancer patients each treated with five fractions were retrospectively analyzed. For each daily MRI set, OLAR and reposition (No-OLAR) plans were created and the superior plan with the daily fraction determined per clinical dose-volume criteria. SSIM and entropy maps were extracted from each daily MRI set, with respect to its reference (e.g., dry-run) MRI in the region enclosed by 50-100% isodose surfaces. A total of six common features were extracted from SSIM maps. Pearson's rank correlation coefficient was utilized to rule out redundant SSIM features. A t-test was used to determine significant SSIM features which were combined with the change in entropy to develop anensemble machine classifier with fivefold cross validation. The performance of the classifier was evaluated using the area under the curve (AUC) of the receiver operating characteristic curve. A machine learning classifier model using two SSIM features (mean and full width at half maximum) and change in entropy was determined to be able to significantly discriminate between No-OLAR and OLAR groups. The obtained machine learning ensemble classifier can predict OLAR necessity with a cross validated AUC of 0.93. Misclassification was found primarily for No-OLAR cases with dosimetric plan quality closely comparable to the corresponding OLAR plans, thus, are not a major practical concern. A machine learning classifier based on simple first-order image features, that is, SSIM features and change in entropy, was developed to determine when OLAR is necessary for a daily MRI set with practical acceptable prediction accuracy. This classifier may be implemented in the MRgART process to automatically and objectively determine if OLAR is required following daily MRI.


Therapeutic Drug Monitoring of Tacrolimus-Personalized Therapy: Second Consensus Report.
Ten years ago, a consensus report on the optimization of tacrolimus was published in this journal. In 2017, the Immunosuppressive Drugs Scientific Committee of the International Association of Therapeutic Drug Monitoring and Clinical Toxicity (IATDMCT) decided to issue an updated consensus report considering the most relevant advances in tacrolimus pharmacokinetics (PK), pharmacogenetics (PG), pharmacodynamics, and immunologic biomarkers, with the aim to provide analytical and drug-exposure recommendations to assist TDM professionals and clinicians to individualize tacrolimus TDM and treatment. The consensus is based on in-depth literature searches regarding each topic that is addressed in this document. Thirty-seven international experts in the field of TDM of tacrolimus as well as its PG and biomarkers contributed to the drafting of sections most relevant for their expertise. Whenever applicable, the quality of evidence and the strength of recommendations were graded according to a published grading guide. After iterated editing, the final version of the complete document was approved by all authors. For each category of solid organ and stem cell transplantation, the current state of PK monitoring is discussed and the specific targets of tacrolimus trough concentrations (predose sample C0) are presented for subgroups of patients along with the grading of these recommendations. In addition, tacrolimus area under the concentration-time curve determination is proposed as the best TDM option early after transplantation, at the time of immunosuppression minimization, for special populations, and specific clinical situations. For indications other than transplantation, the potentially effective tacrolimus concentrations in systemic treatment are discussed without formal grading. The importance of consistency, calibration, proficiency testing, and the requirement for standardization and need for traceability and reference materials is highlighted. The status for alternative approaches for tacrolimus TDM is presented including dried blood spots, volumetric absorptive microsampling, and the development of intracellular measurements of tacrolimus. The association between CYP3A5 genotype and tacrolimus dose requirement is consistent (Grading A I). So far, pharmacodynamic and immunologic biomarkers have not entered routine monitoring, but determination of residual nuclear factor of activated T cells-regulated gene expression supports the identification of renal transplant recipients at risk of rejection, infections, and malignancy (B II). In addition, monitoring intracellular T-cell IFN-g production can help to identify kidney and liver transplant recipients at high risk of acute rejection (B II) and select good candidates for immunosuppression minimization (B II). Although cell-free DNA seems a promising biomarker of acute donor injury and to assess the minimally effective C0 of tacrolimus, multicenter prospective interventional studies are required to better evaluate its clinical utility in solid organ transplantation. Population PK models including CYP3A5 and CYP3A4 genotypes will be considered to guide initial tacrolimus dosing. Future studies should investigate the clinical benefit of time-to-event models to better evaluate biomarkers as predictive of personal response, the risk of rejection, and graft outcome. The Expert Committee concludes that considerable advances in the different fields of tacrolimus monitoring have been achieved during this last decade. Continued efforts should focus on the opportunities to implement in clinical routine the combination of new standardized PK approaches with PG, and valid biomarkers to further personalize tacrolimus therapy and to improve long-term outcomes for treated patients.