Fatty acid composition and δ13 C of bulk and individual fatty acids as marker for authenticating Italian PDO/PGI extra virgin olive oils by means of isotopic ratio mass spectrometry.
European Regulation (EEC) 2568/91 has been setting the minimum requirements in order to allow labeling of oil as extra virgin. These general requirements, are based on physical-chemical and organoleptic parameters directly linked to the freshness and quality of the product. Isotope ratio mass spectrometry (IRMS) was demonstrated to be a useful tool for the discrimination of the origin of unknown samples, because the obtained data are practically independent of the cultivar employed and the production technique. In this work, the evaluation of the composition of fatty acid methyl esters (FAME) alongside with the determination of stable isotope ratio of C in bulk oils and in main FAME constituents have been investigated as a tool to improve geographical discrimination of Italian Protected Designation of Origin/Protected Geographical Indication (PDO/PGI) samples. For this purpose, authentic PDO/PGI extra virgin olive oils were sampled at oil mills and grouped into different sets according to their areas of provenience. The use of principal component analysis and partial least squares discriminant analysis multivariate analysis techniques demonstrated that discrimination of olive oil samples can be done using geographical and pedoclimatic parameters predominantly by using δ(13) C results of bulk and individual fatty acids. Results showed that δ(13) C values are a more reliable marker of origin with respect to fatty acid composition.


Chitosan coatings crosslinked with genipin for corrosion protection of AZ31 magnesium alloy sheets.
In this study, coatings of chitosan crosslinked with genipin were prepared on sheets of AZ31 magnesium alloy and their corrosion protection properties were characterized by means of potentiodynamic polarization and electrochemical impedance spectroscopy (EIS). The coatings were also characterized by means of FTIR and XPS. It was observed that the crosslinking process decreases the corrosion current and shifts the corrosion potential of the alloy to less negative values. The EIS analysis demonstrated that the crosslinking process increases the maximum impedance after short and long exposure times. The superior performance of the crosslinked coatings is related to a lower degree of swelling, as observed in the swelling tests carried out on free-standing films.


[Mechanical circulatory supports. Advanced stage of heart failure].
Heart failure is considering one of the most prevalent diseases in recent years. Despite advances in the treatment with new drugs and devices such as implantable defibrillator and cardiac resynchronization therapy, the progression in heart failure leads to deterioration in the quality of life, frequent hospitalizations and high mortality rates. Advanced stage of this disease remains heart transplantation for treatment choice, although is not the adequate therapy for all the patients. Many of them require support with inotropic drugs, which lead to a progressive deterioration and limited survival, because waiting times are prolonged or transplantation is not performed. Therefore, progress in circulatory support devices allow to consider these as the treatment while awaiting transplantation and an alternative when the transplant is not considered an option. It is our objective to review the progress and the usefulness of circulatory support devices in patients with advanced heart failure, in which the last option is heart transplantation.


BubR1 recruitment to the kinetochore via Bub1 enhances spindle assembly checkpoint signaling.
During mitosis, unattached kinetochores in a dividing cell activate the spindle assembly checkpoint (SAC) and delay anaphase onset by generating the anaphase-inhibitory mitotic checkpoint complex (MCC). These kinetochores generate the MCC by recruiting its constituent proteins, including BubR1. In principle, BubR1 recruitment to signaling kinetochores should increase its local concentration and promote MCC formation. However, in human cells BubR1 is mainly thought to sensitize the SAC to silencing. Whether BubR1 localization to signaling kinetochores by itself enhances SAC signaling remains unknown. Therefore, we used ectopic SAC activation (eSAC) systems to isolate two molecules that recruit BubR1 to the kinetochore, the checkpoint protein Bub1 and the KI and MELT motifs in the kinetochore protein KNL1, and observed their contribution to eSAC signaling. Our quantitative analyses and mathematical modeling show that Bub1-mediated BubR1 recruitment to the human kinetochore promotes SAC signaling and highlight BubR1's dual role of strengthening the SAC directly and silencing it indirectly.


Next-generation multiple myeloma treatment: a pharmacoeconomic perspective.
Advances in the diagnosis and treatment of multiple myeloma have come at a rapid pace, especially with several new drugs entering the market in the last few years. However, access to and affordability of new treatments poses a major challenge, both in the United States and around the world. High costs of life-saving drugs are detrimental to both the personal finances of the individual patient, as well as society which must bear the increasing costs in terms of increased health insurance premiums, taxes, or both. The challenges are not unique to myeloma, but are commonly encountered in several other cancers as well. But to some extent these pharmacoeconomic concerns are amplified in myeloma due to the need for multidrug regimens that combine 2 or more expensive new drugs, continuous therapy, and the prolonged disease course in most patients. We examine current myeloma therapy from a pharmacoeconomic perspective, and discuss the costs involved. We outline the underlying reasons why cancer drugs are so expensive, the measures that are required to lower cost, and propose potential ways in which costs can be reduced while still delivering high-quality care.


Comparison of Conventional and Standardized Bone Densitometry around Implants in Periapical Radiographs during a Three Months Period.
Comparing continuous films taken at different timescales is a way to study the alveolar bone changes around the implant over time. One of the important concerns in quantitative analysis of the alveolar bone changes over the time is to reduce variations in the X-ray imaging geometry and image density. Using a modified XCP film holder together with the bite recording material, parallel periapical radiographs were taken from the implants placements of 16 patients in four steps. Densities of radiographs were measured in a conventional way using the video densitometry device. The same films were also scanned; sequential radiographic density of each patient was homogenised and the density was measured. Density changes obtained in both methods were compared. The data were evaluated using ANOVA, paired t-test and Pearson correlation (α = 0.05). IN THE CONVENTIONAL METHOD OF DENSITOMETRY, THE AVERAGE DENSITIES WERE AS FOLLOWS: before operation 1.0044, after one week 0.9600, after one month 0.9469 and after three months 0.9398. Also, in the standard method of densitometry, the average densities were as follows: before operation 111.7013, after one week 113.4225, after one month 119.4075 and after three months 131.1162. Average density in conventional densitometry were not significantly different in various time stages (P = 0.395). But, the standard densitometry method showed a significant difference (P = 0.001). The average density obtained at different stages in the standard densitometry showed a gradual increase in the bone density in the entire process. Standardising the patient's consecutive radiographic images is essential for quantitative measurements over the time.


Effects of nuclear factor kappa B signaling pathway in human intervertebral disc degeneration.
IL-1β (interleukin-1β) can activate human nucleus pulposus cells with or without nuclear factor kappa B (NF-κB) inhibition. We undertook a descriptive and mechanistic investigation of catabolic effects of NF-κB signaling pathway in intervertebral disc degenerative changes. To clarify the mediatory role of NF-κB signaling pathway in human intervertebral disc degeneration (IDD). IDD is a major cause of lower back pain, but the molecular mechanism behind this process is poorly understood. NF-κB is a family of transcription factors that play a central role in mediating cellular response to damage, stress, and inflammation. Growing evidence implicates chronic activation of NF-κB in many degenerative diseases, but its role in IDD has not been adequately explored. Human nucleus pulposus cells in monolayer culture were exposed to IL-1β, which increases matrix-degrading enzyme activity in the nucleus pulposus, with or without NF-κB inhibition by BAY11-7082; ribonucleic acid was isolated for real-time polymerase chain reaction analysis of gene expression, Western blot analysis was performed to detect the changes of protein expression. NF-κB specific inhibitor BAY11-7082 significantly inhibited IL-1β-induced NF-κB activation. IL-1β-dependent gene upregulation of matrix metalloproteinase (MMP)-3, MMP-9, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4, and ADAMTS-5 was significantly reduced by NF-κB inhibition. The decreased gene expression of aggrecan and type II collagen, induced by IL-1β was also reversed by BAY11-7082. NF-κB inhibition reversed the IL-1β-induced changes of protein expression of MMP-3, MMP-9, MMP-13, ADAMTS-4, ADAMTS-5, aggrecan, and type II collagen. These findings demonstrate that the NF-κB signaling pathway is a key mediator of IDD and represents a therapeutic target for mitigating disc degenerative diseases. N/A.


The Caenorhabditis elegans unc-13 gene product is a phospholipid-dependent high-affinity phorbol ester receptor.
The Caenorhabditis elegans unc-13 mutant is a member of a class of mutants that have un-coordinated movement. Mutations of the unc-13 gene cause diverse defects in C. elegans, including abnormal neuronal connections and modified synaptic transmission in the nervous system. unc-13 cDNA encodes a protein (UNC-13) of 1734 amino acid residues with a predicted molecular mass of 198 kDa and sequence identity to the C1/C2 regions but not to the catalytic domain of the ubiquitously expressed protein kinase C family [Maruyama & Brenner (1991) Proc. Natl. Acad. Sci. U.S.A. 88, 5729-5733]. To characterize the phorbol ester binding site of the UNC-13 protein, cDNA encoding the C1/C2-like regions (amino acid residues 546-940) was expressed in Escherichia coli and the 43 kDa recombinant protein was purified. Phorbol ester binding to the 43 kDa protein was zinc- and phospholipid-dependent, stereospecific and of high affinity (Kd 67 nM). UNC-13 specific antisera detected a protein of approx. 190 kDa in wild-type (N2) but not in mutant (e1019) C. elegans cell extracts. We conclude that UNC-13 represents a novel class of phorbol ester receptor.


Rescue of a submerged convulsing diver.
In 2018, the Medical Panel of the NATO Underwater Diving Working Group (UDWG) discussed the question of the rescue and management of a submerged unresponsive compressed-gas diver. The Panel reviewed the 2012 recommendation by the UHMS Diving Committee with respect to the specific recommendation in a convulsing diver using a half-face mask and separate mouthpiece, to delay surfacing until the clonic phase had subsided if the mouthpiece was in place. There is a paucity of scientific, epidemiological, experimental and observational human studies to substantiate this guidance. Experimental animal studies suggest that the likelihood of a complete airway obstruction during an ongoing seizure is low and that there is a high likelihood of surviving pulmonary barotrauma caused by complete airway closure. Airway management and control is an essential step in the management of the unresponsive diver and would be challenging to achieve in the underwater environment. Even in the military setting, it will be difficult to provide sufficient training to enable divers to handle such a situation. In this very rare scenario it is considered that emergency guidelines should be clear, concise and easy to follow. The UDWG therefore recommends that all unconscious military divers in this situation should be rescued to surface without waiting for clonic seizures to subside. Training organizations for recreational and occupational divers should consider whether this guidance should be applied for civilian divers as well.


CONVERSION OF LEUCOANTHOCYANINS INTO THE CORRESPONDING ANTHOCYANIDINS.
Purified preparations of leucoanthocyanins from several sources were heated in butanol-hydrochloric acid solutions from 50 degrees to 90 degrees C and the rate of production of cyanidin was measured. From the temperature dependence of this rate, the calculated energy of activation was of the order of magnitude of 20,000 calories. The rate-limiting step in this reaction was similar for two preparations tested, and of lower energy for a third, presumably of lower degree of polymerization.


One Step Closer to Safer: Counseling Outcomes from American Academy of Pediatrics Firearm Safe Storage Education Training.
To assess pre and postself-reported counseling outcomes for Safer: Storing Firearms Prevents Harm, an American Academy of Pediatrics universal firearm safe storage counseling training for pediatric clinicians providing health maintenance in outpatient settings. Safer was developed by a national multidisciplinary committee of firearm injury prevention experts with input from firearm-owning families and launched in June 2021. Pediatric clinicians completed baseline and 1-month posttraining surveys after signing up for the Pedialink course from February through June 2022. Primary outcomes included self-reported measures of counseling self-efficacy and frequency. Wilcoxon matched-pairs signed-rank tests compared outcome distributions at baseline and follow-up. Two adjusted, multilevel mixed-effects regression models were conducted. Of 230 clinicians who completed baseline surveys, 146 (64%) completed 1-month postsurveys. Regional representation included Southeast = 67 (46%), Northeast = 24 (16%), Midwest = 21 (14%), Pacific = 15 (10%), Southwest = 11 (8%), and Rocky Mountain = 8 (5%). At follow-up, there was significant improvement in both the distribution of self-efficacy (median [first Quartile-third Quartile = 50 [20-70] at baseline and 80 [60-85] at follow-up; P < .001) and self-reported counseling frequency (median [first Quartile-third Quartile] = 10 [0-50] at baseline and 50 [10-80] at follow-up; P < .001). Adjusted regression model results suggested that self-efficacy significantly improved from baseline to follow-up (time coefficient 25.3; 95% CI = [21.0, 29.5]; P < .001) as did counseling frequency (time coefficient 13.6; 95% CI = [9.2, 18.0]; P < .001). Significant improvement in self-reported counseling self-efficacy and frequency was demonstrated 1 month following Safer training.


Occupational risks of bladder cancer in the United States: I. White men.
We examined the relationship between occupation and bladder cancer risk using data obtained from interviews conducted with 2,100 white males with bladder cancer and 3,874 population controls during the National Bladder Cancer Study, a population-based, case-control study conducted in 10 areas of the United States. The strongest evidence of increased risk among white men was observed for painters, truck drivers, and drill press operatives. For painters, the overall relative risk was 1.5 [95% confidence intervals (CI) = 1.2-2.0]. Among painters who started working prior to 1930, a significant trend in risk with increasing duration of employment as a painter was apparent; the relative risk for such painters employed 10 or more years was 3.0. For truck drivers and drill press operatives, overall risks were 1.3 (CI = 1.1-1.4) and 1.4 (CI = 0.9-2.1), respectively. We observed a significant, positive trend in risk with increasing duration of employment in each of these occupations, with relative risks peaking at approximately two for long-term workers. Excess risks were also observed for workers in several other occupations. In all, we estimate that 21%-25% of bladder cancer diagnosed among white men in the United States is attributable to occupational exposures.


Environmentally responsive plasmonic nanoassemblies for biosensing.
Assemblies of plasmonic nanoparticles enable new modalities for biosensing. Engineered superstructures from metal nanoparticles can enhance the plasmon resonances and chiroptical activity of nanoscale dispersions. Such phenomena are keys to the fabrication of highly sensitive, selective and fast-responding detection platforms, making them promising candidates for clinical applications. This tutorial review summarizes and discusses recent advances in this area. The topics covered in the review include the basic strategies adopted for assembly and engineering of plasmonic nanoparticles, optical properties of the assembled nanostructures and their applications to both in vitro and in vivo detection of biological compounds. We also offer our vision of the future prospects of this field of research. Among emerging applications in this area are novel nanosensors and platforms, for food safety, environmental monitoring, health safeguarding, as well as biodefense.


A Review on Mental Stress Assessment Methods Using EEG Signals.
Mental stress is one of the serious factors that lead to many health problems. Scientists and physicians have developed various tools to assess the level of mental stress in its early stages. Several neuroimaging tools have been proposed in the literature to assess mental stress in the workplace. Electroencephalogram (EEG) signal is one important candidate because it contains rich information about mental states and condition. In this paper, we review the existing EEG signal analysis methods on the assessment of mental stress. The review highlights the critical differences between the research findings and argues that variations of the data analysis methods contribute to several contradictory results. The variations in results could be due to various factors including lack of standardized protocol, the brain region of interest, stressor type, experiment duration, proper EEG processing, feature extraction mechanism, and type of classifier. Therefore, the significant part related to mental stress recognition is choosing the most appropriate features. In particular, a complex and diverse range of EEG features, including time-varying, functional, and dynamic brain connections, requires integration of various methods to understand their associations with mental stress. Accordingly, the review suggests fusing the cortical activations with the connectivity network measures and deep learning approaches to improve the accuracy of mental stress level assessment.


Patient-level outcomes and virologic suppression rates in HIV-infected patients receiving antiretroviral therapy in Rwanda.
The Rwanda national HIV program has been successful at scaling up antiretroviral therapy (ART) to achieve universal access. The AIDSRelief Model of Care focuses on four key principles: (1) earlier initiation of ART; (2) use of durable, highly-potent, and sequence-friendly first-line ART regimens; (3) early detection of treatment failure; and (4) provision of community-based care and support to ensure optimal adherence and follow up/engagement in care. We conducted a retrospective cohort study of randomly-selected HIV-infected patients at AIDSRelief-supported sites using a stratified, random sample of 583 adults (>15 years) who initiated ART from 30 June 2008 to 1 February 2010. At ART initiation, the median patient age was 38 years, and 67% were female. The baseline median CD4+ cell count was 309 cells/mm3. Overall virologic suppression was 91%. Married/ever married status (adjusted prevalence odds ratio [aPOR] 3.75, 95% confidence interval [CI] 1.30-10.78) and self-reported adherence ≥95% in the past month (aPOR 2.76, 95% CI 1.00-7.62) were significantly associated with viral suppression in the multivariable model. Excellent virologic outcomes were achieved in Rwandan AIDSRelief sites utilizing the AIDSRelief Model of Care during the scale-up of ART in the country.


Chemical Composition of East Asian Invasive Knotweeds, their Cytotoxicity and Antimicrobial Efficacy Against Cariogenic Pathogens: An In-Vitro Study.
BACKGROUND Giant knotweeds originating from East Asia, such as Reynoutria japonica, and Reynoutria sachalinensis, and their hybrid such as Reynoutria x bohemica, are invasive plants in Europe and North America. However, R. japonica is also a traditional East Asian drug (Polygoni cuspidati rhizoma) used in Korean folk medicine to improve oral hygiene. The aim of this study was to evaluate the antibacterial activity of acetone extracts of Reynoutria species against dominant caries pathogen such as Streptococcus mutans and alternative pathogens, as well as characterize the phytochemical composition of extracts and examine their cytotoxicity. MATERIAL AND METHODS Ultrasonic extraction was used to obtain polyphenol-rich extracts. The extracts were characterized by HPLC-DAD-ESI-MS. To test bacterial viability, the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) against S. mutans, S. salivarius, S. sanguinis, and S. pyogenes were determined. The cytotoxicity of the extracts to human fibroblasts derived from gingiva was evaluated using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. RESULTS The R. japonica extract had the highest bacteriostatic and bactericidal activity against pathogens causing caries, mainly dominant caries pathogen S. mutans (mean MIC 1000 μg/mL and MBC 2000 μg/mL), which was most likely associated with a higher content of stilbene aglycons and anthraquinone aglycons in the extract. Moreover, the R. japonica extract demonstrated the lowest cytotoxic effect on human fibroblasts and exhibited cytotoxic activity only at the concentration causing the death of all S. mutans. CONCLUSIONS The results indicate that the R. japonica acetone extract can be considered as a natural, antimicrobial agent for caries control.


Characteristics and Outcomes of Physician-to-Physician Telephone Consultation Programs: Environmental Scan.
Telephone consultations between physicians provide quick access to medical advice, allowing patients to be cared for by calling physicians in their local settings. As part of a quality assurance study of a physician-to-physician consultation program in Alberta, Canada, this environmental scan aims to identify the characteristics and outcomes of physician-to-physician telephone consultation programs across several countries. We searched 7 databases to identify English publications in 2007-2017 describing physician-to-physician consultations using telephones as the main technology. To identify Canadian programs, the literature search was supplemented with an additional internet search. The literature search yielded 2336 citations, of which 17 publications were included. Across 7 countries, 14 telephone consultation programs provided primary care providers with access to various specialists through hotlines, paging systems, or call centers. The programs reported on the avoidance of hospitalizations, emergency department visits and specialty visits, caller satisfaction with the telephone consultation, and cost avoidance. Telephone consultation programs between health care providers have facilitated access to specialist care and prevented acute care use. .


Studies on the in situ nitrogen degradability corrected for bacterial contamination of concentrate feeds in steers.
Stable 15N was used to evaluate the influence of bacterial contamination on in situ DM and N degradabilities (Dg) of meat and bone meal (MBM), soybean meal (SBM), and wheat bran (WB) in two steers. Bacterial DM and N contamination ranged from 2.4 to 28.6% and 2.1 to 56.8% of residual DM and N, respectively. Effective N degradability increased when bacterial contamination was taken into account (P < .05). The difference was low for MBM (2.4%) and for SBM (3.4%) but high for WB (12.2%). Theoretically, using solid-associated bacteria should give the most accurate correction for bacterial contamination; however, results showed that Dg of N based on liquid-associated bacteria were identical for MBM and SBM (P > .05) and different for WB (P < .05). In a second experiment, five treatments were applied to incubated feeds to remove bacteria fixed to the residues and consequently to determine directly the Dg of DM and N corrected for the bacterial contamination without the need for a marker. These treatments involved chilling for 6 h at 4 degrees C in saline solution alone (T1) or with a commercial detergent (T2), or with sodium dodecyl sulfate (T3) or with methylcellulose (T4), followed by pummeling in a stomacher for 5 min. The last treatment was only machine washing twice (T5). The Dg of DM can be directly determined following the first four treatments, nevertheless their application to MBM and SBM led to higher Dg of N than that corrected for the bacterial contamination determined in the first experiment (P < .05).(ABSTRACT TRUNCATED AT 250 WORDS)


Growth curves of clients' emotional experience and their association with emotion regulation and symptoms.
Emotional experience during psychotherapy is considered a core mechanism of change. Yet the sheer experience itself may not necessarily be beneficial; instead, the trajectories of emotional experience need to be explored as possible predictors of treatment outcomes. This study investigated whether clients' pre-treatment levels of emotion regulation and symptoms predicted patterns of session-to-session change in emotional experience. We also explored which patterns better predict clients' improvement in emotion regulation and symptoms from pre- to post treatment. One-hundred and seven clients undergoing psychodynamic psychotherapy completed questionnaires on their symptoms and emotion regulation at pre- and post- treatment. They also reported their level of emotional experience at the end of each session. Pre-treatment symptoms and difficulties in emotion regulation predicted greater instability in emotional experience. Higher mean levels of emotional experience during treatment were associated with an improvement in emotion regulation, and greater stability during treatment was associated with improvement in emotion regulation and symptoms. These findings lend weight to the idea that experiencing emotion in the therapeutic environment has significant implications for clients' ability to manage their emotions outside the session. However, emotions experienced in an unstable manner within therapy are associated with poorer outcomes. Clinical and methodological significance of this article: Therapists can benefit from observing the patterns and not only the level of their clients' emotional experiences. The identification of clients' difficulties early in treatment may help therapists guide clients through the delicate process of carefully attending to their emotions.


Continuous, on-line DNA sequencing using oligodeoxynucleotide primers with multiple fluorophores.
A method for sequencing DNA by using a difluoresceinated primer and laser excitation is described. Dideoxy protocols have been determined that provide sequences for 600 bases starting with base 1 with less than 1% error in a single load. Electrophoresis is at 20 W and the bands are detected 24 cm from the bottom of the loading well with a scanning fluorescence detector. Bands are imaged on a TV screen in two dimensions. The sequences can be read from the TV screen manually or semiautomatically by using a simple software program. The system allows more bases to be read with a lower error rate than any other reported automated sequencing method.


Magnetic and structural relationship of RFe2Si2 and R(Fe(1-x)M(x))2Si2(x = 0-1) systems (R = La, Y and Lu, M = Ni, Mn and Cu).
Due to the similarity between AFe(2)As(2) (A = Ba, Sr) and RFe(2)Si(2) (R = La, Y and Lu), the RFe(2)Si(2) system has been proposed as a potential candidate for a new high TC superconducting family containing Fe-Si (instead of Fe-As) layers as a structural unit. Various R (Fe(1-x)M(x))2 Si(2) (M = Ni and Cu) materials were synthesized and measured for their magnetic properties. None of these materials is superconducting down to 1.8 K. A pronounced peak at 232 K was observed in the magnetization curve of YFe(2)Si(2). (57)Fe Mössbauer studies confirm the absence of any long-range magnetic ordering below 232 K. Similar peaks at various temperatures also appear in R (Fe(1-x)M(x))2 Si2 samples. For Y (Fe(1-x)Mn(x))2 Si(2) the peak position is dramatically affected by the magnetic Mn dopants. Four independent factors affect the peak position and shift it to lower temperatures: (i) the lattice parameters, (ii) the concentration of x (iii) the applied magnetic field and (iv) the magnetic nature of M. It is proposed that the magnetic peaks observed in RFe(2)Si(2) and in R (Fe(1-x)M(x))2 Si(2) represent a new nearly ferromagnetic Fermi liquid system, its nature is yet to be determined.


Perspectives on PARP inhibitors as pharmacotherapeutic strategies for breast cancer.
Introduction Approximately 10% of all breast cancer cases occur in individuals who have germline pathogenic variants of the BRCA 1, BRCA 2, and other genes associated with impaired DNA damage repair that is associated with an increased risk of breast, ovarian, and other cancers. Inhibitors of poly-ADP ribose polymerase (PARP) induce synthetic lethality in cancer cells harboring such pathogenic variants.Area covered In this review, the authors review the mechanisms of action, antitumor activity, and adverse events associated with PARP inhibitors for the treatment of advanced breast cancer. The authors then summarize the area and provide their expert perspectives on the area.Expert opinion Two PARP inhibitors are approved in metastatic breast cancer, including olaparib and talozaparib. Both agents were approved based on phase III trials demonstrating that they were associated with improved progression-free survival compared with treatment of physician's choice in patients receiving second-third line therapy for locally advanced, inoperable, or metastatic breast cancer in patients with germline pathogenic BRCA 1 or BRCA2 variants.


Expression and polarized targeting of a rab3 isoform in epithelial cells.
Pathways of polarized membrane traffic in epithelial tissues serve a variety of functions, including the generation of epithelial polarity and the regulation of vectorial transport. We have identified a candidate regulator of polarized membrane traffic in epithelial cells (i.e., rab3B), which is a member of the rab family of membrane traffic regulators. Rab3B is highly homologous to a brain-specific rab3 isoform (rab3A) that targets in a polarized fashion to the presynaptic nerve terminal, where it probably regulates exocytosis. The coding region for human rab3B was cloned from epithelial mRNA using a reverse-transcription polymerase chain reaction strategy. This cDNA clone hybridized to a single mRNA species in Northern blots of poly(A)+ RNA isolated from epithelial cell lines. A rab3B-specific antibody that was raised against recombinant fusion protein recognized a 25-kD band in immunoblots of cell lysates prepared from cultured epithelial cells (e.g., T84 and HT29-CL19A), but not from a variety of nonepithelial cells (e.g., PC12 neuroendocrine cells). Immunofluorescence analysis confirmed that rab3B protein is preferentially expressed in cultured epithelial cells as well as in a number of native epithelial tissues, including liver, small intestine, colon, and distal nephron. Rab3B localized to the apical pole very near the tight junctions between adjacent epithelial cells within all of these cell lines and native epithelial tissues, as determined by immunofluorescence and immunoelectron microscopic analysis. Moreover, this pattern of intracellular targeting was regulated by cell contact; namely, rab3B was reversibly retrieved from the cell periphery as epithelial cell contact was inhibited by reducing the extracellular Ca2+ concentration. Our results indicate that neurons and epithelial cells express homologous rab3 isoforms that target in a polarized fashion within their respective tissues. The pattern and regulation of rab3B targeting in epithelial cells implicates this monomeric GTPase as a candidate regulator of apical and/or junctional protein traffic in epithelial tissues.


Health care reform: Part I.
The provision of access to quality health care services and the restraint of rapidly escalating health care costs are dominant policy issues. This article traces governmental and private efforts to develop a health insurance program and enact various reforms from the beginning of the 20th century to 1980. Problems and challenges that influence the current debate will be shown to have existed over the past several decades.


Altered interactions among thin filament proteins modulate cardiac function.
The transition of cardiac muscle myofilament activity from the diastolic to the systolic state is a complex allosteric/co-operative process, rich in potential control mechanisms. The rate and intensity of the transition is modulated by the mechanical state of the myofilaments, by covalent and non-covalent mechanisms, and by the isoform population of myofilament proteins. Moreover, the process is altered in pathological states and subject to modification by pharmacological agents with potential use as inotropic drugs. We present here a current perception of the process, with focus on molecular interactions of the thin filament components, especially troponin I. Our discussion is couched in terms of what we believe to be pressing questions in the current state of knowledge of this system. These questions are as follows: what is the topology of the thin filament and how do thin filament proteins regulate the activation of cross-bridge cycling? What is the relative role of protein phosphorylation of thin filament proteins in the regulation of the cardiac activity and dynamics? What is the relative role of feedback effects of cross-bridge binding on thin filament activity? Answers to these questions have taken on new significance, with the identification of familial hypertrophic cardiomyopathy as a "sarcomeric" disease related to missense mutations in myosin, troponin T. and tropomyosin. As discussed, new and exciting developments in this area are bringing us closer to the answers to these questions.


Effectiveness and Characterization of Severely Energy-Restricted Diets in People with Class III Obesity: Systematic Review and Meta-Analysis.
Severely energy-restricted diets are used in obesity management, but their efficacy in people with class III obesity (body mass index ≥40 kg/m2) is uncertain. The aims of this systematic review and meta-analysis were to determine the effectiveness and characteristics of severely energy-restricted diets in people with class III obesity. As there was a lack of publications reporting long-term dietary interventions and randomised controlled trial designs, our original publication inclusion criteria were broadened to include uncontrolled study designs and a higher upper limit of energy intake. Eligible publications reported studies including adults with class III obesity and that assessed a diet with daily energy intake ≤5000 kJ for ≥4 weeks. Among 572 unique publications from 4 databases, 11 were eligible and 10 were suitable for meta-analysis. Our original intention was to classify comparison arms into short-term (<6 months) and long-term (>1 year) interventions. Due to the lack of long-term data found, comparison arms were classified according to the commonalities in dietary intervention length among the included publications, namely dietary interventions of 4 weeks' duration and those of ≥6 weeks' duration. After a 4-week severely energy-restricted diet intervention, the pooled average weight loss was 9.81 (95% confidence interval 10.80, 8.83) kg, with a 95% prediction interval of 6.38 to 13.25 kg, representing a loss of approximately 4.1 to 8.6% of initial body weight. Diets ≥6 weeks' duration produced 25.78 (29.42, 22.15) kg pooled average weight loss, with a 95% prediction interval of 13.77 to 37.80 kg, representing approximately 10.2 to 28.0% weight loss. Daily dietary prescriptions ranged from 330 to 5000 kJ (mean ± standard deviation 2260 ± 1400 kJ), and had wide variations in macronutrient composition. The diets were administered mostly via liquid meal replacement products. While the included publications had a moderate risk of bias score, which may inflate reported weight loss outcomes, the published data to date suggest that severely energy-restricted diets, delivered via diets of varying composition, effectively produce clinically relevant weight loss (≥10% of initial body weight) when used for 6 weeks or more in people with class III obesity.


European Network of Forensic Science Institutes (ENFSI): Evaluation of new commercial STR multiplexes that include the European Standard Set (ESS) of markers.
To support and to underpin the European initiative to increase the European set of standard markers (ESS), by the addition of five new loci, a collaborative project was organised by the European Network of Forensic Science Institutes (ENFSI) DNA working group in order to assess the new multiplex kits available. We have prepared allele frequency databases from 26 EU populations. Concordance studies were carried out to verify that genotyping results were consistent between kits. Population genetics studies were conducted and it was estimated that F(ST)<0.001. The results showed that the kits were comparable to each other in terms of performance and major discrepancy issues were highlighted. We provide details of allele frequencies for each of the populations analysed per laboratory.


The neural basis of logopenic progressive aphasia.
Logopenic progressive aphasia (LPA) is defined clinically by impairments of naming and sentence repetition. The relationship between these impairments and their neural basis has, however, not yet been determined. We aimed to localize cortical thinning associated with naming and repetition deficits using cortical thickness measurements. Consecutive LPA cases (n = 15) were matched with healthy controls (n = 16). All LPA cases underwent general cognitive testing and language assessment using the progressive aphasia language scale. Word retrieval and verbal short-term memory, the core cognitive processes involved in LPA, were assessed using visual confrontation naming and forward digit-span tasks. Cortical thickness was estimated vertex-by-vertex using Freesurfer. The pattern of cortical thinning for the LPA group as well as the location of cortical thinning linked to the impairment of each core cognitive process was estimated using general linear models. LPA cases showed extensive left-sided cortical thinning in which the temporo-parietal junction had the greatest involvement. Impaired naming was associated with cortical thinning of the supramarginal gyrus (BA 40), while reduced digit-span score, regarded as a surrogate marker for sentence repetition, was correlated with thinning of the left superior temporal gyrus (BA 22 and 42). These results suggest that the core manifestations of LPA emerge from the damage to segregated and non-overlapping cortical regions typically affected in this focal presentation of Alzheimer's disease.


The in vitro elution characteristics of antifungal-loaded PMMA bone cement and calcium sulfate bone substitute.
The use of antimicrobial-loaded delivery vehicles, most often as antibiotic beads, is common practice for the treatment of deep musculoskeletal infections. The elution of antibacterial drugs from various bone cements has been extensively studied. However, much less is known about the elution of other antimicrobials from these materials. In particular, the use of this approach for fungal infections has not been well studied despite growing concern about these difficult-to-treat organisms. Voriconazole is a broad-spectrum and highly effective antifungal that has been used in the treatment of resistant fungal pathogens. We examined the in vitro elution characteristics of voriconazole from nonabsorbable polymethylmethacrylate (PMMA) beads and from absorbable calcium sulfate beads. Voricanazole-containing beads were immersed in a 5-mL bath of phosphate-buffered saline at room temperature and placed on an orbital shaker. Eluent samples were collected over the course of 2 weeks. Concentrations of the antifungal drug in solution were measured using high-performance liquid chromatography. To verify biologic activity of the eluted antifungal, collected samples were also tested against control yeasts. We found that samples collected out to 2 weeks contained relatively high voriconazole concentrations and enough active antifungal activity to inhibit growth of the control yeasts. These data demonstrate that voriconazole retains its antifungal activity when mixed into either PMMA or calcium sulfate beads, and elutes out of beads at biologically effective concentrations over a time period of at least 2 weeks. Therefore, incorporation of voriconazole into either absorbable or nonabsorbable beads appears to be a reasonable strategy for the local delivery of a potent, broad-spectrum antifungal agent to an infected wound bed.


Adult stem cells and mammalian epimorphic regeneration-insights from studying annual renewal of deer antlers.
Mammalian organ regeneration is the "Holy Grail" of modern regenerative biology and medicine. The most dramatic organ replacement is known as epimorphic regeneration. To date our knowledge of epimorphic regeneration has come from studies of amphibians. Notably, these animals have the ability to reprogram phenotypically committed cells at the amputation plane toward an embryonic-like cell phenotype (dedifferentiation). The capability of mammals to initiate analogous regeneration, and whether similar mechanisms would be involved if it were to occur, remain unclear. Deer antlers are the only mammalian appendages capable of full renewal, and therefore offer a unique opportunity to explore how nature has solved the problem of mammalian epimorphic regeneration. Following casting of old hard antlers, new antlers regenerate from permanent bony protuberances, known as pedicles. Studies through morphological and histological examinations, tissue deletion and transplantation, and cellular and molecular techniques have demonstrated that antler renewal is markedly different from that of amphibian limb regeneration (dedifferentiation-based), being a stem cell-based epimorphic process. Antler stem cells reside in the pedicle periosteum. We envisage that epimorphic regeneration of mammalian appendages, other than antler, could be made possible by recreating comparable milieu to that which supports the elaboration of that structure from the pedicle periosteum.


Analysis of microsatellite instability (MSI) in pediatric gonadal and extra-gonadal germ cell tumors.
Gonadal and extragonadal pediatric germ cell tumors (GCTs) are rare neoplasms with different clinical behavior. Although surgery and cisplatin-based chemotherapy are resolutive in most cases, some patients do not respond to chemotherapy and have a worse outcome. Microsatellite instability (MSI) was correlated to resistance to chemotherapy and sensitivity to immunotherapy in different neoplasms. A series of 21 pediatric GCTs were tested by immuno-histochemistry and PCR to evaluate MSI status. Next generation sequencing was applied to further evaluate cases with discordant results between immunohistochemistry and PCR. Twenty-one cases of pediatric GCT were included in the series. The mean age ranged between 1 and 10 years. Nine cases were gonadal GCTs and the remaining 12 were extra-gonadal GCTs. By immunohistochemistry, one case showed a deficit of Mismatch repair (MMR) proteins. This case was a 1-year-old children affected by gonadal yolk sac tumor. However, all cases resulted microsatellite stable (MSS) by PCR and NGS. MSI was not detected in our series of pediatric GCTs, as well as the data present in literature about adult patients with GCTs. Molecular techniques could have a role to confirm the MSI status in case of dMMR by immunohistochemistry.


Fracture risk in adult patients treated with growth hormone replacement therapy for growth hormone deficiency: a prospective observational cohort study.
To our knowledge, no controlled studies of the effects of long-term growth hormone replacement on fracture risk in adult patients with growth hormone deficiency exist. We assessed the effect of growth hormone treatment on fracture risk in patients with growth hormone deficiency from the international Hypopituitary Control and Complications Study (HypoCCS) surveillance database. In this prospective cohort study, patients with growth hormone deficiency were analysed from the HypoCCS database of adults with hypopituitarism from the USA, Canada, Japan, and 14 European countries. Patients were eligible if they were aged 18 years or older and had an established diagnosis of growth hormone deficiency, either alone or with multiple pituitary hormone deficiencies, as identified by clinical history and biochemical testing. Patients were assessed over a mean follow-up period of 4·6 years (SD 3·8). The effect of growth hormone treatment on fracture risk was assessed by Cox proportional hazard modelling with adjustment for several confounders. Between Jan 3, 1996, and Dec 15, 2012, we enrolled 10,673 patients to this study. Of the enrolled patients, 1032 patients were excluded from assessment because of incomplete data, leaving 9641 in the analysis cohort. Of these patients, 8374 of received growth hormone and 1267 did not. Annual fracture incidence rate was lower in patients who received growth hormone than in those who did not (fracture incidence rate 1·19% vs 1·91%, hazard ratio [HR] 0·69, 95% CI 0·54-0·88). However, no difference in fracture risk was observed between patients who did and did not receive growth hormone treatment in the subgroup of patients with pre-existing osteoporosis (n=826; 0·97, 0·48-1·95). Our results suggest that growth hormone replacement therapy could be protective against fracture for adult patients with growth hormone deficiency without previously reported osteoporosis. Starting growth hormone therapy before the onset of osteoporosis might be optimum for bone health of adult patients with growth hormone deficiency. Eli Lilly and Co.


T1-weighted MR imaging of the brain using a fast inversion recovery pulse sequence.
The purpose of this paper was to develop and evaluate a fast inversion recovery (FIR) technique for T1-weighted MR imaging of contrast-enhancing brain pathology. The FIR technique was developed, capable of imaging 24 sections in approximately 7 minutes using two echoes per repetition and an alternating echo phase encoding assignment. Resulting images were compared with conventional T1-weighted spin echo (T1SE) images in 18 consecutive patients. Compared with corresponding T1SE images, FIR images were quantitatively comparable or superior for lesion-to-background contrast and contrast-to-noise ratio (CNR). Gray-to-white matter and cerebrospinal fluid (CSF)-to-white matter contrast and CNR were statistically superior in FIR images. Qualitatively, the FIR technique provided comparable lesion detection, improved lesion conspicuity, and superior image contrast compared with T1SE images. Although FIR images had greater amounts of image artifacts, there was not a statistically increased amount of interpretation-interfering image artifact. FIR provides T1-weighted images that are superior to T1SE images for a number of image quality criteria.


[Methylation of prokaryotic RNA by S-methyl methionine in in vivo experiments].
While cultivating the E. coli 113-3 strain on the mineral medium containing S-methyl-(methyl-3H)-methionine, the incorporation of methyl groups into 4S, 16S and 23S RNA proved to be over 5 times more effective as compared with the control, when L-(methyl-3H)-methionine acted as a donor of methyl groups. The ratio of methylated components has much in common and significant differences. All the types of RNA of E. coli grown in the mineral medium containing S-methyl methionine showed an increased ratio of m2G as compared with the corresponding types of RNA of E. coli after its cultivation in the methionine containing mineral medium.


Nuclear factor kappaB- and specificity protein 1-dependent p53-mediated bi-directional regulation of the human manganese superoxide dismutase gene.
Tumor suppressor p53 is known to activate certain sets of genes while suppressing others. However, whether p53 can both activate and suppress the same gene is unclear. To address this question, concentration-dependent p53 effect on the manganese superoxide dismutase (MnSOD) gene was investigated. By transfecting p53 in PC-3 cells, we demonstrate that low concentrations of p53 increase while high concentrations suppress MnSOD expression. The physiological relevance of this effect was determined in vitro and in vivo using combined UVB-mediated activation and small interference RNA-mediated suppression of p53. Results were consistent with the bi-directional effect of p53 on MnSOD expression. MnSOD-promoter/enhancer analysis demonstrates that p53 is suppressive to the promoter activity regardless of the presence or absence of putative p53 binding sites. However, a low level of p53 increases MnSOD gene transcription in the presence of the intronic-enhancer element, and this effect is dependent on nuclear-factor kappaB (NF-kappaB) binding sites. Expression of p53 enhances nuclear levels of p65 with corresponding increase in the DNA-binding activity of NF-kappaB as detected by electrophoretic mobility shift and chromatin immunoprecipitation assays. Transfection of p65 small interference RNA reduces the positive effect of p53 on MnSOD gene transcription. These data suggest that p65 can overcome the negative effect of p53 on MnSOD expression. However, when the level of p53 was further increased, the suppressive effect of p53 outweighed the positive effect of p65 and led to the suppression of MnSOD gene transcription. These results demonstrated that p53 can both suppress and induce MnSOD expression depending on the balance of promoter and enhancer binding transcription factors.


Cytidine analogs are synthetic lethal with base excision repair default due to MBD4 deficiency.
Inactivating mutations of MBD4 have been reported in subsets of various tumors. A deficiency of this DNA glycosylase, recognizing specifically T:G mismatch resulting from the deamination of methyl-cytosine, results in a hypermutated phenotype due to the accumulation of CpG>TpG transitions. Here, we hypothesize that the difference in DNA metabolism consecutive to MBD4 deficiency may result in specific cytotoxicities in MBD4-deficient tumor cells in a synthetic lethality fashion. After a large-scale drug repurposing screen, we show in two isogenic MBD4 knock-out cell models that the inactivation of MBD4 sensitizes cancer cells to cytidine analogs. We further confirm the exquisite activity of gemcitabine in an MBD4-deficient co-clinical model as (i) it completely prevented the development of an MBD4-deficient uveal melanoma patient-derived xenograft and (ii) treatment in the corresponding patient resulted in an exceptional tumor response. These data suggest that patients harboring MBD4-deficient tumors may be treated efficiently by cytidine analogs.


A study on upper extremity cumulative trauma disorder in different unorganised sectors of West Bengal, India.
Cumulative trauma disorder (CTD) is a term used to describe a class of soft tissue injuries that result due to a number of occupational activities. These disorders commonly occur among workers who are engaged in highly repetitive jobs involving continuous hand exertion, vibration and localized mechanical pressure. In the present investigation, an attempt was made to evaluate the prevalence of CTD among workers associated with strenuous hand intensive jobs in unorganized sectors in India and to highlight the unsafe working conditions to which these workers have been exposed for several years. For this purpose, an experiment was performed on 25 male workers from each group. The groups were classified into meat cutters, typists, tailors, visual display terminal (VDT) operators & weavers. For the symptom survey, a questionnaire and checklist method was implemented. Along with these, a detailed time study was performed among the workers during different activities in the total work cycle. For this study a two-tail chi-square test of independence was applied to determine whether or not the feeling of discomfort had any significant association with the repetitiveness of the work. From the observations and analysis of the results, it was revealed that all the activities are repetitive, i.e. over 50% of the work cycle of each activity involved the respective main activity where similar kinds of motion patterns were performed. Therefore it can be concluded that high repetitiveness, prolonged work activity and remaining in static posture for a prolong period of time may be regarded as the causative factors in the occurrence of CTD.


Correlation between insertion torque and survival rates in immediately loaded implants in the maxilla: a retrospective study.
To evaluate the correlation between insertion torque and the survival rate of immediately loaded implants placed in the maxilla. 390 NobelReplace (Nobel Biocare USA, Yorba Linda, CA) Select Tapered implants were placed in the maxillae of 145 patients over a 15-month period. The insertion torque was measured using a digital torque wrench. Of the 390 implants, only 9 (2.3%) failed. The 381 successful implants were placed with an average insertion torque of 72.1 N.cm. Insertion torque values of failed implants varied between 45.8 and 134 N.cm, with an average of 69.2 N.cm. Of the 9 failed implants, 8 were abutments in multiple-unit implant-supported prostheses, and 1 was a single-unit implant. Although insertion torque undoubtedly plays an important role in the primary stability and successful osseointegration of implants, there seems to be no correlation between insertion torque and implant failure above 35 N.cm threshold of insertion torque. Immediate loading of implants in the maxilla is a successful and predictable treatment option with failure rates approaching those of traditional 2-stage surgery.


The role of sleepiness on arterial stiffness improvement after CPAP therapy in males with obstructive sleep apnea: a prospective cohort study.
Obstructive sleep apnea (OSA) is associated with increased cardiovascular risk. This study aim to assess differences in changes in arterial stiffness of two groups of patients, defined as having daytime sleepiness or not, after continuous positive airway pressure (CPAP) treatment. A selected cohort of consecutive male patients, under 65 years old, with moderate to severe OSA and without great number of comorbidities was studied. The diagnosis was confirmed by home respiratory poligraphy. Sleepiness was considered with an Epworth Sleepiness Scale (ESS) > 10. An ambulatory blood pressure (BP) monitoring and carotid-femoral pulse wave velocity (cf-PWV) measurements were performed, before and after four months under CPAP. Compliant patients, sleepy and non-sleepy, were compared using linear mixed effects regression models. A further stratified analysis was performed with non-sleepy patients. Thirty-four patients were recruited, with mean age 55.2 (7.9) years, 38.2% were sleepy, 79.4% with hypertension, 61.8% with metabolic syndrome and 82.4% with dyslipidaemia. In univariable analysis, cf-PWV was strongly related to systolic BP parameters and age, but also to antihypertensive drugs (p = 0.030), metabolic syndrome (p = 0.025) and daytime sleepiness (p = 0.004). Sleepy patients had a more severe OSA, with AHI 44.8 (19.0) vs 29.7 (15.7) events/h (p = 0.018), but sleep study parameters were not associated with cf-PWV values. On multivariable regression, a significant interaction between time (CPAP) and sleepiness (p = 0.033) was found. There was a weak evidence of a cf-PWV reduction after CPAP treatment (p = 0.086), but the effects of treatment differed significantly between groups, with no changes in non-sleepy patients, while in sleepy patients a significant decrease was observed (p = 0.012). Evaluating non-sleepy patients group under CPAP therapy, results showed that both higher pulse pressure (p = 0.001) and lower LDL-cholesterol levels (p = 0.015) at baseline were associated to higher cf-PWV changes. Patients with daytime sleepiness had a more severe OSA and presented a greater arterial stiffness improvement after CPAP therapy, independently from age and BP. Besides sleepiness, cf-PWV reduction after CPAP therapy was mainly associated to CV risk factors, and less to sleep study parameters. Clinicaltrials.gov NCT02273089 23.10.2014 retrospectively registered.


Arabidopsis and Nicotiana anthocyanin production activated by maize regulators R and C1.
Anthocyanin pathway-specific transcriptional activators R and C1 from the monocot maize were expressed in two dicots, Arabidopsis thaliana and Nicotiana tabacum. Expression of R caused augmented anthocyanin pigmentation in both plant species and augmented trichome (hair) production in Arabidopsis. Alone, C1 had no effect. Hybrid transgenic Arabidopsis expressing both C1 and R produced anthocyanins in root, petal, and stamen tissues that normally never express anthrocyanins. When R was expressed in the transparent testa glabrous (without anthocyanins and trichomes) mutant of Arabidopsis, the deficiency was complemented and both anthocyanins and trichomes were restored.


Mitochondrial creatine kinase isoform expression does not correlate with its mode of action.
In adult mammalian ventricular tissue, mitochondrial creatine kinase (mi-CK), which is bound to the outer surface of the mitochondrial inner membrane, is functionally coupled to oxidative phosphorylation. This is shown, in saponin-permeabilized rat ventricular fibres, by a shift in the apparent K(m) of mitochondrial respiration for ADP from 300 +/- 56 microM to 111 +/- 40 microM (P < 0.001) on the addition of 25 mM creatine, due to a local accumulation of ADP close to the ATP/ADP translocator. We have found that in atrial fibres, the apparent K(m) for ADP is high, but is not decreased by creatine, suggesting an absence of coupling in this tissue, as has previously been observed in smooth muscle. mi-CK is encoded by two different genes, which are expressed in a tissue-specific manner: the sarcomeric isoform is expressed in ventricular and skeletal muscles, while the ubiquitous isoform is expressed in smooth muscle, brain and other tissues. In order to determine whether a specific function can be attributed to the expression of a specific isoform, we investigated mi-CK mRNA expression by Northern blot analysis. Hybridization with synthetic oligonucleotides specific for each mi-CK isoform showed the expression of only the sarcomeric isoform in rat atria. This result was confirmed by PCR using primers specific for each isoform. In addition, electrophoretic analysis of CK isoforms showed no difference in the octamer/dimer ratio of mi-CK in the atria and ventricles. In atria, unlike the soleus or ventricles, the maximum potential rate of mitochondrial phosphocreatine synthesis was lower than the maximal rate of ATP production by the mitochondria. The total CK/adenylate kinase ratio was also lower in atria than in the other tissues, suggesting a greater contribution of adenylate kinase to adenine nucleotide compartmentation in this tissue. The functional differences between mi CK in the two cardiac tissues seem to imply a specific arrangement of the proteins in the intermembrane space rather than the expression of specific isoforms.


An ingenious one-dimensional zirconium phosphonate with efficient strontium exchange capability and moderate proton conductivity.
A new 1-D zirconium phosphonate [(CH3)2NH2]2[Zr(CH2(HPO3)(PO3))2] (SZ-5) was synthesized via a solvothermal reaction and its single crystal structure was elucidated. SZ-5 exhibits efficient strontium exchange capability with high uptake capacity and selectivity, as further demonstrated by the radioactive Sr-90 removal from a real contaminated seawater sample with an extremely high ionic strength. In addition, the measured proton conductivity at 90 °C and 90% relative humidity (RH) is 5.65 × 10-4 S cm-1. The efficient ion-exchange ability and the moderate proton conductivity suggest the potential applications of SZ-5 in fuel cells or in the remediation of contaminated water.


Bone mineral density in children and adolescents with systemic lupus erythematosus, juvenile dermatomyositis, and systemic vasculitis: relationship to disease duration, cumulative corticosteroid dose, calcium intake, and exercise.
To describe the frequency of abnormal bone mineralization in a population of children with juvenile systemic lupus erythematosus (JSLE), juvenile dermatomyositis (JDM), and systemic vasculitis; and to investigate the relationship of bone mineral density (BMD) to cumulative corticosteroid dose, disease duration, Tanner stage, calcium intake, and exercise in these patients. A retrospective chart review of children attending the pediatric rheumatology clinic at British Columbia's Children's Hospital was conducted to obtain demographic data (sex, ethnicity, disease duration, cumulative corticosteroid dose, and mean daily corticosteroid dose). All patients had at least one BMD measurement by dual energy x-ray absorptiometry (DEXA) at lumbar spine, hip, and total body. BMD was expressed as g/cm2 and Z scores; an abnormal Z score was defined as </= -1.5. Daily calcium intake was calculated. Physical activity was scored using a validated 7 day physical activities questionnaire. A total of 36 patients were studied. Twenty-five patients had SLE, 7 had JDM, and 4 had systemic vasculitis. Fourteen subjects were Caucasian, 13 Asian, 6 East Indian, and 3 Canadian First Nations. An abnormal Z score at one or more sites was found in 10/25 (40%) with JSLE, and in 3/11 (27%) JDM/vasculitis patients. The mean Z scores (+/- SD) for lumbar spine were -1.02 (+/- 1.2), for hip -0.88 (+/- 1.3), and for total body -0.77 (+/- 1.5). Compared to children with normal bone densities, those with lower bone density tended to be younger (13.5 +/- 2.2 vs 15.5 +/- 1.7 yrs), received higher corticosteroid dosages at the time of the study (0.78 +/- 0.6 vs 0.35 +/- 0.2 mg/kg), and were more often prepubertal (OR for total body scores 5, 95% CI 0.7-46). Decreased bone density is common in children with SLE and other systemic rheumatic diseases. Age, corticosteroid dose, and pubertal stage all appear to have some influence on bone mass in these children.


Calcium absorptive efficiency is positively related to body size.
Calcium absorption efficiency is a more important determinant of calcium balance than calcium intake itself. The sources of variability in absorptive performance are only partly elucidated. The aim of the study was to explore the relationship between body size and calcium absorption efficiency. Metabolic studies were performed on an inpatient metabolic unit in an academic health sciences center. One hundred seventy-eight women, with an average age of 50.2 yr, were studied from one to five times and yielded an aggregate data set containing 633 individual studies. Calcium absorption fraction was measured by the dual-tracer method. Observed values were expressed as residuals from predicted values for each woman's actual calcium intake, using the previously published relationship between intake and absorption. Absorption residuals were significantly positively correlated with height, weight, and surface area, and after adjusting for estrogen status, these body size variables accounted for approximately 4% of the total variability. The magnitude of the effect is such that a woman 1.8 m in height would absorb 30+% more calcium from a given intake than a woman 1.4 m tall.


[Compulsive buying or oniomania: an overview].
Compulsive buyers feel a continual urge to buy things, irrespective of their financial means. Interest in compulsive buying has increased in the last 30 to 40 years. Since excessive buying is a regularly occurring feature of various illnesses encountered in clinical practice, one wonders whether it should be regarded as a sign of the times, as a symptom of a psychiatric illness or as a distinct psychiatric syndrome. To gain insight into the concept of compulsive buying. We searched the literature via Medline, PubMed and PsycInfo for the period from 1966 to August 2008. results Compulsive buying is a relatively unknown and poorly defined phenomenon, despite its high incidence and high comorbidity. It is characterised by chronic, repetetive behaviour; it occurs mainly among women and should be classified as an impulsive-control disorder nos. Current aetiological models lack explanatory power and, so far, the most successful treatment results seem to have been achieved with behavioural therapy. Compulsive buying should be considered from a neuroscientific perspective. Future research needs to have a neuropsychiatric focus and a contextual psychological and behavioural analytical approach in which inhibition, self-control and rule-governed behaviour play a central role.


New approaches for studying and exploiting an old protuberance, the plant trichome.
Much recent study of plant trichomes has focused on various aspects of glandular secreting trichomes (GSTs) and differentiation of simple trichomes. This Botanical Briefing will highlight: research on various aspects of, and manipulation of glandular secreting trichomes; molecular aspects of the differentiation and development of simple trichomes of arabidopsis and cotton; how methods for manipulation of model systems used in the above work can be applied to expand our understanding of less studied surface structures of plants. The Briefing will cover: established and suggested roles of simple and glandular secreting trichomes; recent results regarding solute and ion movement in trichomes; methods for isolating trichomes; recent studies of trichome differentiation and development; attempts to modify metabolism in secreting trichomes; efforts to exploit trichomes for commercial and agronomic purposes.


Moisture Stress as a Requirement for Flowering of Coffee.
When coffee plants are watered at relatively short intervals, so as to maintain the water content of the soil at close to field capacity, the flower buds remain dormant and no fruits are formed. Irrigation or rain induces flowering only when preceded by a period of water shortage. Water stress is apparently essential to break the dormancy of coffee flower buds.


Association of the Ras-antagonistic Rap1/Krev-1 proteins with the Golgi complex.
Ras oncogenes encode 21-kDa GTP-binding proteins that are capable of transforming immortalized cells in culture. Ras proteins are bound to the inner face of the plasma membrane by their C-terminal extremity and are thought to transmit their mitogenic signals via an "effector" domain spanning amino acids 32-42. Two ras-related human genes rap1A and rap1B encode 95% homologous 21-kDa proteins that share with Ras p21 the same effector domain and a similar C-terminal Cys-Ali-Ali-Xaa sequence (where Ali is an aliphatic amino acid; also known as a CAAX sequence). The product of the rap1A gene is identical to that of the Krev-1 cDNA, whose overexpression is capable of reverting the phenotype of Ki-ras-transformed NIH 3T3 cells. Antibodies that do not cross-react with Ras and other Ras-related proteins were obtained by immunizing rabbits with a peptide encompassing residues 121-137 of Rap1 proteins. These antibodies were used to investigate the subcellular localization of Rap1 proteins by indirect immunofluorescence and fractionation techniques. Rap1 proteins were found to be tightly bound to cellular membranes. They did not colocalize with Ras proteins on the plasma membrane and were discovered to be associated with the Golgi complex.


Nephronectin is Correlated with Poor Prognosis in Breast Cancer and Promotes Metastasis via its Integrin-Binding Motifs.
Most cancer patients with solid tumors who succumb to their illness die of metastatic disease. While early detection and improved treatment have led to reduced mortality, even for those with metastatic cancer, some patients still respond poorly to treatment. Understanding the mechanisms of metastasis is important to improve prognostication, to stratify patients for treatment, and to identify new targets for therapy. We have shown previously that expression of nephronectin (NPNT) is correlated with metastatic propensity in breast cancer cell lines. In the present study, we provide a comprehensive analysis of the expression pattern and distribution of NPNT in breast cancer tissue from 842 patients by immunohistochemical staining of tissue microarrays from a historic cohort. Several patterns of NPNT staining were observed. An association between granular cytoplasmic staining (in <10% of tumor cells) and poor prognosis was found. We suggest that granular cytoplasmic staining may represent NPNT-positive exosomes. We found that NPNT promotes adhesion and anchorage-independent growth via its integrin-binding and enhancer motifs and that enforced expression in breast tumor cells promotes their colonization of the lungs. We propose that NPNT may be a novel prognostic marker in a subgroup of breast cancer patients.


Use of photopatterned porous polymer monoliths as passive micromixers to enhance mixing efficiency for on-chip labeling reactions.
We introduce a passive micromixer with novel architecture using photopatterned porous polymer monoliths (PPM) and demonstrate an improvement in mixing efficiency by monitoring the fluorescence of an on-chip labeling reaction. UV light was used to photopattern a periodic arrangement of PPM structures directly within the channel of a plastic microfluidic chip. By optimizing the composition of the polymerization solution and irradiation time we demonstrate the ability to photopattern PPM in regularly repeating 100 microm segments at the tee-junction of the disposable device. To evaluate the efficiency of this dual functional mixer-reactor fluorescamine and lysine were introduced in separate channels upstream of the tee-junction and the intensity of laser-induced fluorescence resulting from the fluorogenic labeling reaction was monitored. The fluorescence level after the photopatterned periodic monolith configuration was 22% greater than both an equivalent 1 cm continuous segment of PPM and an open channel. Results indicate that this periodic arrangement of PPM, with regularly spaced open areas between 100 microm plugs of PPM, is directly responsible for enhancing the mixing and overall rate of chemical reaction in the system. In addition to facilitating preparation of a dual functional mixer-reactor, the ability to accurately photopattern PPM is an enabling technology for seamlessly integrating multiple monoliths into a single device. This technology will be particularly important to proteomic applications requiring preconcentration, enzymatic digestion and two-dimensional separations.


Functional repression of cAMP response element in 6-hydroxydopamine-treated neuronal cells.
Impaired survival signaling may represent a central mechanism in neurodegeneration. 6-Hydroxydopamine (6-OHDA) is an oxidative neurotoxin used to injure catecholaminergic cells of the central and peripheral nervous systems. Although 6-OHDA elicits phosphorylation of several kinases, downstream transcriptional effects that influence neuronal cell death are less defined. The cAMP response element (CRE) is present in the promoter sequences of several important neuronal survival factors. Treatment of catecholaminergic neuronal cell lines (B65 and SH-SY5Y) with 6-OHDA resulted in repression of basal CRE transactivation. Message levels of CRE-driven genes such as brain-derived neurotrophic factor and the survival factor Bcl-2 were decreased in 6-OHDA-treated cells, but message levels of genes lacking CRE sequences were not affected. Repression of CRE could be reversed by delayed treatment with cAMP several hours after initiation of 6-OHDA injury. Furthermore, restoration of CRE-driven transcription was associated with significant neuroprotection. In contrast to observations in other model systems, the mechanism of CRE repression did not involve decreased phosphorylation of its binding protein CREB. Instead, total CREB and phospho-CREB (pCREB) were increased in the cytoplasm and decreased in the nucleus of 6-OHDA-treated cells. 6-OHDA also decreased nuclear pCREB in dopaminergic neurons of primary mouse midbrain cultures. Co-treatment with cAMP promoted/restored nuclear localization of pCREB in both immortalized and primary culture systems. Increased cytoplasmic pCREB was observed in degenerating human Parkinson/Lewy body disease substantia nigra neurons but not in age-matched controls. Notably, cytoplasmic accumulation of activated upstream CREB kinases has been observed previously in both 6-OHDA-treated cells and degenerating human neurons, supporting a potential role for impaired nuclear import of phosphorylated signaling proteins.


TATDN1 promotes the development and progression of breast cancer by targeting microRNA-140-3p.
To explore whether long non-coding RNA (lncRNA) TATDN1 can promote the proliferation and cell cycle progression of breast cancer cells by adsorbing microRNA-140-3p, thus participating in the development of breast cancer (BCa). Expressions of TATDN1 and microRNA-140-3p in BCa tissues and paracancerous tissues were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Meanwhile, TATDN1 expression in BCa cell lines was detected as well. Regulatory effects of TATDN1 and microRNA-140-3p on proliferation and cell cycle progression of BCa cells were evaluated by Cell Counting Kit-8 (CCK-8) and flow cytometry, respectively. The binding relationship of microRNA-140-3p to NOVA1 and TATDN1 was examined by dual-luciferase reporter gene assay. Finally, rescue experiments were conducted to explore whether TATDN1 can regulate NOVA1 expression by adsorbing microRNA-140-3p to exert its biological function in BCa. TATDN1 was highly expressed in BCa tissues and cell lines. Upregulation of TATDN1 promoted the proliferative potential and cell cycle progression of MCF-7 and MDA-MB-231 cells. Dual-luciferase reporter gene assay indicated that TATDN1 could bind to microRNA-140-3p, which was lowly expressed in BCa. Overexpression of microRNA-140-3p inhibited the proliferative potential and cell cycle progression of MCF-7 and MDA-MB-231 cells. Moreover, microRNA-140-3p partially inhibited the role of TATDN1 in regulating cellular behaviors of BCa cells. NOVA1 was predicted to be the target gene of microRNA-140-3p. Overexpression of NOVA1 partially abolished the inhibitory effects of microRNA-140-3p on proliferation and cell cycle progression of MCF-7 and MDA-MB-231 cells. TATDN1 promotes the proliferative potential and cell cycle progression of BCa cells through adsorbing microRNA-140-3p to upregulate NOVA1 expression.


Promoter methylation of HIN-1 in the progression to esophageal squamous cancer.
HIN-1 (High in normal-1) is a tumor suppressor gene that is highly expressed in many epithelial tissues, including breast lung, trachea, pancreas, prostrate and salivary gland. Inactivation of HIN-1 expression by promoter methylation is frequent in many epithelial carcinomas and carcinoma-in-situ, including breast, lung and nasopharyngeal. Because HIN-1 silencing commences at an early stage of malignant transformation in these tissues, it may be a useful marker for tumor presence. The status of HIN-1 regulation in esophageal cancer has not been previously reported. Thus, we analyzed 18 esophageal cancer cell lines for HIN-1 expression and methylation by reverse transcription PCR (RT-PCR) and methylation specific PCR (MSP). HIN-1 gene silencing and promoter methylation was present in 13 (72%) of the cell lines. Bisulfite-treated sequencing confirmed the methylation status in cell lines and demonstrated dense methylation of HIN-1 throughout the promoter region. Epigenetic changes of HIN-1 were examined throughout the progression of carcinogenesis in esophageal squamous lesions through analysis of archived surgical specimens from patients with normal esophageal mucosa (n = 17), grade I dysplasia (n = 39), grade II dysplasia (n = 12), grade III dysplasia (n = 9), and invasive squamous esophageal cancer (n = 126). Methylation of HIN-1 was present in 0% of normal mucosa, 31% of grade I dysplasia, 33% of grade II dysplasia, 44% of grade III dysplasia, and 50% of esophageal cancer specimens analyzed. These studies demonstrate HIN-1 silencing is associated with dense promoter region hypermethylation in esophageal cancer and suggest that methylation of HIN-1 is an early event in dysplastic transformation.


Genetic variations and humoral immune responses to myelin oligodendroglia glycoprotein in adult phenotypes of X-linked adrenoleukodystrophy.
The lack of phenotype/genotype association in X-linked adrenoleukodystrophy (X-ALD) has prompted the search for disease modifying factors. We previously demonstrated increased serum antibody responses against myelin oligodendrocyte glycoprotein (MOG) in various clinical phenotypes of X-ALD allowing speculations that myelin specific humoral immune responses might be involved in phenotype generation of X-ALD. In the present study, we investigated the possible association of (1) a naturally occurring variable number tandem repeat (vntr) polymorphism (C allele) in the 3' flanking region of the interleukin-6 gene (IL-6), previously demonstrated to modify the course of Alzheimer's disease, systemic lupus erythematodes and Multiple Sclerosis (MS), (2) a tetranucleotide repeat polymorphism (TAAA)(n) in the 3' flanking region of the MOG gene and (3) HLA class II alleles with adult clinical phenotypes and serum antibody responses to MOG in 70 adult X-ALD patients. HLA class II alleles, (TAAA)(n) MOG gene polymorphisms, and IL-6 C allele were not associated with clinical phenotypes. Anti-MOG antibodies were detectable in 17/54 X-ALD patients (31.5%). Anti-MOG antibodies were associated with the 226 bp (TAAA)(n) MOG gene polymorphism but not with distinct clinical phenotypes.


Unmet need for mental health care in schizophrenia: an overview of literature and new data from a first-admission study.
We present an overview of the literature on the patterns of mental health service use and the unmet need for care in individuals with schizophrenia with a focus on studies in the United States. We also present new data on the longitudinal course of treatments from a study of first-admission patients with schizophrenia. In epidemiological surveys, approximately 40% of the respondents with schizophrenia report that they have not received any mental health treatments in the preceding 6-12 months. Clinical epidemiological studies also find that many patients virtually drop out of treatment after their index contact with services and receive little mental health care in subsequent years. Clinical studies of patients in routine treatment settings indicate that the treatment patterns of these patients often fall short of the benchmarks set by evidence-based practice guidelines, while at least half of these patients continue to experience significant symptoms. The divergence from the guidelines is more pronounced with regard to psychosocial than medication treatments and in outpatient than in inpatient settings. The expansion of managed care has led to further reduction in the use of psychosocial treatments and, in some settings, continuity of care. In conclusion, we found a substantial level of unmet need for care among individuals with schizophrenia both at community level and in treatment settings. More than half of the individuals with this often chronic and disabling condition receive either no treatment or suboptimal treatment. Recovery in this patient population cannot be fully achieved without enhancing access to services and improving the quality of available services. The recent expansion of managed care has made this goal more difficult to achieve.


Selective extraction of triazine herbicides based on a combination of membrane assisted solvent extraction and molecularly imprinted solid phase extraction.
A selective extraction technique based on the combination of membrane assisted solvent extraction and molecularly imprinted solid phase extraction for triazine herbicides in food samples was developed. Simazine, atrazine, prometon, terbumeton, terbuthylazine and prometryn were extracted from aqueous food samples into a hydrophobic polypropylene membrane bag containing 1000μL of toluene as the acceptor phase along with 100mg of MIP particles. In the acceptor phase, the compounds were re-extracted onto MIP particles. The extraction technique was optimised for the type of organic acceptor solvent, amount of molecularly imprinted polymers particles in the organic acceptor phase, extraction time and addition of salt. Toluene as the acceptor phase was found to give higher triazine binding onto MIP particles compared to hexane and cyclohexane. Extraction time of 120min and 100mg of MIP were found to be optimum parameters. Addition of salt increased the extraction efficiency for more polar triazines. The selectivity of the technique was demonstrated by extracting spiked cow pea and corn extracts where clean chromatograms were obtained compared to only membrane assisted solvent extraction or only molecularly imprinted solid phase extraction. The study revealed that this combination may be a simple way of selectively extracting compounds in complex samples.


Tsukamurella tyrosinosolvens sp. nov.
Chemotaxonomic and 16S ribosomal DNA sequence analyses of four bacterial isolates from blood cultures from patients with cardiac pacemaker implants and sputa of patients with chronic lung infections clearly demonstrated that these bacteria belong to the genus Tsukamurella. DNA-DNA hybridization data, as well as the physiological characteristics of the isolates, indicate that they are closely related and belong to a single species that differs from previously described members of the genus Tsukamurella. The name Tsukamurella tyrosinosolvens sp. nov. is proposed for these isolates, and the new species is represented by strain IMMIB D-1397T (= DSM 44234T). Strain IMMIB D-1397T exhibits 53.4, 53.5, and 54.7% DNA-DNA relatedness to Tsukamurella paurometabola DSM 20162T, Tsukamurella inchonensis DSM 44067T, and Tsukamurella pulmonis DSM 44142T, respectively.


[Female prostate gland].
The review presents the results of a systematic literature on evaluation of the female prostate gland, its morphology and physiology available in Russian libraries and the U.S. National Library of Medicine.


Targeting synthetic lethality in DNA damage repair pathways as an anti-cancer strategy.
During the process of tumorigenesis, certain cancers are known to develop deficiencies in one or more major pathways of DNA damage repair, rendering them critically dependent on alternative repair processes for maintaining genomic integrity and viability. Targeting these alternative DNA repair mechanisms is a potentially highly-specific anti-cancer strategy, as their inhibition is theoretically toxic only to tumor cells and not to normal tissues. We will review here the rationale behind this strategy and provide examples of its application. We will also discuss several as yet unanswered questions surrounding this strategy, including whether human cancers frequently harbor synthetically lethal interactions in DNA repair and, if so, how patients might be identified who would benefit from targeting such interactions.


Influence of environmental parameters on movements and habitat utilization of humpback whales (Megaptera novaeangliae) in the Madagascar breeding ground.
Assessing the movement patterns and key habitat features of breeding humpback whales is a prerequisite for the conservation management of this philopatric species. To investigate the interactions between humpback whale movements and environmental conditions off Madagascar, we deployed 25 satellite tags in the northeast and southwest coast of Madagascar. For each recorded position, we collated estimates of environmental variables and computed two behavioural metrics: behavioural state of 'transiting' (consistent/directional) versus 'localized' (variable/non-directional), and active swimming speed (i.e. speed relative to the current). On coastal habitats (i.e. bathymetry < 200 m and in adjacent areas), females showed localized behaviour in deep waters (191 ± 20 m) and at large distances (14 ± 0.6 km) from shore, suggesting that their breeding habitat extends beyond the shallowest waters available close to the coastline. Males' active swimming speed decreased in shallow waters, but environmental parameters did not influence their likelihood to exhibit localized movements, which was probably dominated by social factors instead. In oceanic habitats, both males and females showed localized behaviours in shallow waters and favoured high chlorophyll-a concentrations. Active swimming speed accounts for a large proportion of observed movement speed; however, breeding humpback whales probably exploit prevailing ocean currents to maximize displacement. This study provides evidence that coastal areas, generally subject to strong human pressure, remain the core habitat of humpback whales off Madagascar. Our results expand the knowledge of humpback whale habitat use in oceanic habitat and response to variability of environmental factors such as oceanic current and chlorophyll level.


A phosphatase cascade by which rewarding stimuli control nucleosomal response.
Dopamine orchestrates motor behaviour and reward-driven learning. Perturbations of dopamine signalling have been implicated in several neurological and psychiatric disorders, and in drug addiction. The actions of dopamine are mediated in part by the regulation of gene expression in the striatum, through mechanisms that are not fully understood. Here we show that drugs of abuse, as well as food reinforcement learning, promote the nuclear accumulation of 32-kDa dopamine-regulated and cyclic-AMP-regulated phosphoprotein (DARPP-32). This accumulation is mediated through a signalling cascade involving dopamine D1 receptors, cAMP-dependent activation of protein phosphatase-2A, dephosphorylation of DARPP-32 at Ser 97 and inhibition of its nuclear export. The nuclear accumulation of DARPP-32, a potent inhibitor of protein phosphatase-1, increases the phosphorylation of histone H3, an important component of nucleosomal response. Mutation of Ser 97 profoundly alters behavioural effects of drugs of abuse and decreases motivation for food, underlining the functional importance of this signalling cascade.


Effects of Stamm gastrostomy on gastric emptying rate in rats.
Although frequency of gastroesophageal reflux (GER) increases after gastrostomy, the role of gastric emptying in GER has not been evaluated. In this study, we examined the effects of Stamm gastrostomy on gastric emptying rate in rats and whether Stamm gastrostomy induces GER or not. Sprague-Dawley rats were divided into three groups. Stamm gastrostomy was done in the first group (SG). Sham operation was carried out in group 2 and the 3rd group served as control. Gastric emptying was assessed using both liquid and solid meals in each group at postoperative 14th day. For solid meal emptying, after fasting of 16 h, the rats were fed for 3 h and gastric emptying rate was measured at the fifth hour. Methylcellulose was used for emptying of liquids and it was given after the animals were fasted for 16 h and gastric emptying rate was measured 30 min later. Histological evaluation for GER was performed in all groups. GER was observed pathophysiologically in 5 of the 7 rats in SG group. Gastric emptying rates of liquid and solid meals were found to be similar in control, SG or sham groups. Surgical gastrostomy does not affect the gastric emptying of solid and liquid meals in rats. Other mechanisms should be considered in the development of GER observed following gastrostomy.


The shame spiral of addiction: Negative self-conscious emotion and substance use.
The bidirectional associations between negative self-conscious emotions such as shame and guilt and substance use are poorly understood. Longitudinal research is needed to examine the causes, consequences, and moderators of negative self-conscious emotions in people who use substances. Using parallel process latent growth curve modeling, we assessed bidirectional associations between shame and guilt and substance use (i.e., number of days in the past 30 used stimulants, alcohol to intoxication, other substances, or injected drugs) as well as the moderating role of positive emotion. Emotions were assessed using the Differential Emotions Scale. The sample included 110 sexual minority cisgender men with biologically confirmed recent methamphetamine use, enrolled in a randomized controlled trial in San Francisco, CA. Participants self-reported emotions and recent substance use behaviors over six time points across 15 months. Higher initial levels of shame were associated with slower decreases in stimulant use over time (b = 0.23, p = .041) and guilt was positively associated with stimulant use over time (β = 0.85, p < .0001). Initial levels of guilt and alcohol use were positively related (b = 0.29, p = .040), but over time, they had a negative relationship (β = -0.99, p < .0001). Additionally, higher initial levels of other drug use were associated with slower decreases in shame over time (b = 0.02, p = .041). All results were independent of depression, highlighting the specific role of self-conscious emotions. Shame and guilt are barriers to reducing stimulant use, and expanded efforts are needed to mitigate the deleterious effects of these self-conscious emotions in recovery from a stimulant use disorder.


Profiling recombinant major birch pollen allergen Bet v 1a and carbamylated variants with CZE and CIEF.
A preparation of recombinant birch pollen allergen of Betula verrucosa isoform 1a (Bet v 1a) containing chemically modified (carbamylated) variants has been analyzed by CZE and CIEF. In CZE, employing a 100 mmol/L MES buffer at pH 6.50, with 0.4 mmol/L tetraethylenepentamine (TEPA) added, allowed for the resolution of 17 protein fractions. The CIEF profiling of the allergen preparation required a combination of a wide-pH-range carrier ampholyte (CA) of pH 3-10 with two narrow-range CAs of pH 5-6 and 5-7. For CIEF, 91 mmol/L of glycine at pH 2.12 and 20 mmol/L of CHES at pH 10.00 were applied as anolyte and catholyte, respectively. The generated pH gradient was nonlinear with a flat slope for pH 4-6, thus providing an improved resolution. In CIEF, up to 18 protein fractions were distinguished as well. The pI of the target allergen Bet v 1a was 4.9 as determined by means of two pI marker compounds flanking the allergen. Relative purity of the target allergen within the preparation containing carbamylated variants was in accordance for both separation systems and varied between 40.7 and 42.8%.


Epidemiology of measles cases, vaccine effectiveness, and performance towards measles elimination in The Gambia.
In 2011, member states of the World Health Organization (WHO) Africa Regional Office (AFRO) resolved to eliminate Measles by 2020. Our study aims to assess The Gambia's progress towards the set AFRO measles elimination target and highlight surveillance and immunisation gaps to better inform future measles prevention strategies. A retrospective review of measles surveillance data for the period 2011-2019, was extracted from The Gambia case-based measles surveillance database. WHO-UNICEF national coverage estimates were used for estimating national level MCV coverage. Measles post campaign coverage survey coverage estimates were used to estimate national measles campaign coverage. One hundred and twenty-five of the 863 reported suspected cases were laboratory confirmed as measles cases. More than half (53.6%) of the confirmed cases have unknown vaccination status, 24% of cases were vaccinated, 52.8% of cases occurred among males, and 72.8% cases were among urban residents. The incidence of measles cases per million population was lowest (0) in 2011-2012 and highest in 2015 and 2016 (31 and 23 respectively). The indicator for surveillance sensitivity was met in all years except in 2016 and 2019. Children aged 5-9 years (Incidence Rate Ratio-IRR = 0.6) and residents of Central River region (IRR = 0.21) had lower measles risk whilst unvaccinated (Adjusted IRR = 5.95) and those with unknown vaccination status (IRR 2.21) had higher measles risk. Vaccine effectiveness was 89.5%. The Gambia's quest to attain measles elimination status by 2020 has registered significant success but it is unlikely that all target indicators will be met. Vaccination has been very effective in preventing cases. There is variation in measles risk by health region, and it will be important to take it into account when designing prevention and control strategies. The quality of case investigations should be improved to enhance the quality of surveillance for decision making.


Novel ribosomal mutations affecting translational accuracy, antibiotic resistance and virulence of Salmonella typhimurium.
Many mutations in rpsL cause resistance to, or dependence on, streptomycin and are restrictive (hyperaccurate) in translation. Dependence on streptomycin and hyperaccuracy can each be reversed phenotypically by mutations in either rpsD or rpsE. Such compensatory mutations have been shown to have a ram phenotype (ribosomal ambiguity), increasing the level of translational errors. We have shown recently that restrictive rpsL alleles are also associated with a loss of virulence in Salmonella typhimurium. To test whether ram mutants could reverse this loss of virulence, we have isolated a set of rpsD alleles in Salmonella typhimurium. We found that the rpsD alleles restore the virulence of strains carrying restrictive rpsL alleles to a level close to that of the wild type. Unexpectedly, three out of seven mutant rpsD alleles tested have phenotypes typical of restrictive alleles of rpsL, being resistant to streptomycin and restrictive (hyperaccurate) in translation. These phenotypes have not been previously associated with the ribosomal protein S4. Furthermore, all seven rpsD alleles (four ram and three restrictive) can phenotypically reverse the hyperaccuracy associated with restrictive alleles of rpsL. This is the first demonstration that such compensations do not require that the compensating rpsD allele has a ribosomal ambiguity (ram) phenotype.


Diffusion-controlled reactions of enzymes. A comparison between Chou's model and Alberty-Hammes-Eigen's model.
In the study of enzyme-catalysed mechanisms, it is often necessary to calculate the upper limit of enzyme reaction, which is usually used as an important criterion to identify whether an assumed enzyme-catalysed mechanism is reasonable or not. Basically, in the existing methods, there are two kinds of models, the Alberty-Hammes-Eigen model and the Chou model, for calculating the upper limit of enzyme reaction. In this paper, an analysis and comparison between these two models are made. It is pointed out that the magnitude of the van der Waals' binding energy between enzyme and substrate molecules will play a key role in deciding whether there is a significant difference or not for the results calculated from these two models. Through such a comparison, the role of the major protein outside the active site of an enzyme molecule also becomes obvious; if the van der Waals' binding energy is very small, the major protein will act like a 'wall', blocking the flow of some substrate molecules to the active site; while if the van der Waals' energy is greater than 3 kT (where k is the Boltzmann constant and T the absolute temperature), the major protein will behave like a 'accelerator', speeding up the flow of the substrate molecules to the active site around the enzyme molecule.


Surface modification of pharmaceutical nanocarriers with ascorbate residues improves their tumor-cell association and killing and the cytotoxic action of encapsulated paclitaxel in vitro.
To evaluate the potential of ascorbate as a novel ligand in the preparation of pharmaceutical nanocarriers with enhanced tumor-cell specific binding and cytotoxicity. Palmitoyl ascorbate was incorporated into liposomes at varying concentrations. A stable formulation was selected based on size and zeta potential measurements. A co-culture of cancer cells with GFP expressing non-cancer cells was used to determine the specificity of palmitoyl ascorbate liposome binding. Liposomes were fluorescently labeled to facilitate analysis by flow cytometry and fluorescence microscopy. The cytotoxic action of palmitoyl ascorbate liposomes against a variety of cell types was assayed using a standard metabolic assay. The cytotoxic effect of a low dose of paclitaxel incorporated in palmitoyl ascorbate liposomes on various cell lines was also determined. Palmitoyl ascorbate liposomes associated preferentially with various cancer cells compared to non-cancer cells in a co-culture model. Palmitoyl ascorbate liposomes exhibited anti-cancer toxicity in numerous cancer cell lines. Furthermore, ascorbate liposomes enhanced the effectiveness of encapsulated paclitaxel compared to paclitaxel encapsulated in 'plain' liposomes. Surface modification of liposomes with ascorbate residues represents a novel way to target and kill certain types of tumor cells and additionally can potentiate the effect of paclitaxel delivered by the liposomes.