Donor-specific transplantation tolerance: the paradoxical behavior of CD4+CD25+ T cells.
To investigate the antigen specificity of regulatory T cells capable of preventing transplant rejection, we have developed two different strategies to achieve tolerance to fully mismatched skin grafts in euthymic mice. A combination of nondepleting Abs targeting CD4, CD8, and CD154 (CD40 ligand) induces dominant transplantation tolerance to fully mismatched skin allografts. Such tolerance is antigen-specific, mediated by regulatory T cells, and can be extended through linked suppression to naïve lymphocytes. The same protocol, when combined with allogeneic bone marrow, enables the development of mixed hematopoietic chimerism and deletional tolerance. Although we cannot exclude that some regulatory T cells may persist in chimeric mice, these cells are insufficient to mediate linked suppression. CD4(+)CD25(+) T cells, whether taken from naïve mice or from mice tolerized through either treatment protocol, were always able to prevent rejection of skin grafts by naïve CD4(+) T cells, and did so with no demonstrable specificity for the tolerizing donor antigens. Such data question whether CD4(+)CD25(+) regulatory T cells alone can account for the antigen specificity of dominant transplantation tolerance.


Worsening of heart failure by coronavirus disease 2019 is associated with high mortality.
Patients with advanced heart failure (HF) with reduced left ventricular ejection fraction (HFrEF) and concurrent coronavirus disease 2019 (COVID-19) might have a higher risk of severe events. We retrospectively studied 16 patients with advanced HFrEF who developed COVID-19 between 1 March and 29 May 2020. Follow-up lasted until 30 September. Ten patients previously hospitalized with decompensated HFrEF were infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during hospitalization. Six patients undergoing ambulatory care at initiation of COVID-19 symptoms were hospitalized because of advanced HFrEF. All patients who experienced worsening of HFrEF due to COVID-19 required higher doses or introduction of additional inotropic drugs or intra-aortic balloon pump in the intensive care unit. The mean intravenous dobutamine dose before SARS-CoV-2 infection in previously hospitalized patients (n = 10) and the median (inter-quartile range) peak intravenous dobutamine dose during SARS-CoV-2 infection in all patients (n = 16) were 2 (0-7) μg/kg/min and 20 (14-20) (P < 0.001), respectively. During follow-up, 56% underwent heart transplantation (n = 2) or died (n = 7). Four patients died during hospitalization from mixed shock consequent to severe acute respiratory syndrome with inflammatory storm syndrome associated with septic and cardiogenic shock during COVID-19. After COVID-19 recovery, two patients died from mixed septic and cardiogenic shock and one from sustained ventricular tachycardia and cardiogenic shock. Five patients were discharged from hospital to ambulatory care. Four were awaiting heart transplantation. Worsening of advanced HF by COVID-19 is associated with high mortality. This report highlights the importance of preventing COVID-19 in patients with advanced HF.


[Diagnosis and management of Imperforated Hymen in an infant].
Imperforated hymen is the most frequent congenital malformation of the female genital tract. Most cases are not investigated in childhood, due to an insufficient genital examination. Its diagnosis and resolution must be early and definitive to avoid subsequent complications. A clinical case of an infant with imperforated himen is presented. To update on genital examination technique necessary to detect this pathology in the newborn and infants, and the proper treatment. 3-months-old infant that consulted due to an increase in bulging volume in the introitus area. In the physical examination, the Valsalva's maneuver was performed allowed the ex pulsion of a homogeneous pearly mass with a smooth surface, between the labia majora, suggesting imperforate hymen. The study was complemented with gynecological ultrasound, which demons trated the presence of hydrocolpos, ruling out other anomalies. A hymenotomy was performed, which allowed for the evacuation of abundant serous material, with no bad smell, and then the hymenectomy was completed by resection of the hymenal membrane. In follow-up monitoring 4 months after the intervention, a widely permeable hymen was found. Neonatal care teams require training on the correct technique of external genital examination of the newborn and infant. Hymenectomy is the technique that allows definitely resolving the condition, avoiding complications.


Dry Eye Disease in Hemifacial Spasm Patients Treated with Botulinum Toxin Type A.
To assess the impact of botulinum toxin type A (BTX-A) on signs and symptoms of dry eye (DE) in affected eye of hemifacial spasm (HFS) patients and to compare the prevalence of DE between affected and non-affected eye in HFS patients. This prospective study included participants with unilateral HFS, who received BTX-A injection as a treatment. The eyes ipsilateral to the spasm side were used as studied eyes and the contralateral eyes were used as controls. The Ocular Surface Disease Index (OSDI) score, tear break-up time (TBUT), corneal fluorescein staining, and Schirmer I test were measured at baseline, 1 and 3 months after BTX-A injection. Fluorescein clearance test (FCT) was evaluated at baseline and at 1 month after BTX-A injection. Thirty-one participants (6 males and 25 females; mean age 61±10 years) were included. The prevalence of DE according to the Asia Dry Eye Society was not significantly different between affected (37.93%) and non-affected eyes (27.6%); P=0.083. At baseline, there was no significant difference in TBUT, Schirmer test, basal tear secretion, presence of delayed tear clearance, and presence of reflex tear secretion between affected and non-affected eyes, while significant difference in Oxford scheme grade was observed (P=0.031). OSDI score, TBUT, Oxford scheme grade, and Schirmer test at 1 month (P=0.817, 0.796, 0.534, 0.556), and 3 months (P=0.803, 0.904, 0.936, 0.684) after BTX-A injection did not significantly change from baseline in affected eyes. FCT results were not significantly different between baseline and at 1-month follow-up in both groups. All findings were corresponding in both naïve and long-term botulinum toxin injection groups. We found no significant effect of BTX-A on signs and symptoms of DE in patients with HFS. Moreover, there was no significant association between HFS and DE. However, we found significant corneal surface damage in the affected eyes, which emphasized importance of ocular surface evaluation and prompt treatment in HFS patients.


In vivo properties of the disaggregase function of J-proteins and Hsc70 in Caenorhabditis elegans stress and aging.
Protein aggregation is enhanced upon exposure to various stress conditions and aging, which suggests that the quality control machinery regulating protein homeostasis could exhibit varied capacities in different stages of organismal lifespan. Recently, an efficient metazoan disaggregase activity was identified in vitro, which requires the Hsp70 chaperone and Hsp110 nucleotide exchange factor, together with single or cooperating J-protein co-chaperones of classes A and B. Here, we describe how the orthologous Hsp70s and J-protein of Caenorhabditis elegans work together to resolve protein aggregates both in vivo and in vitro to benefit organismal health. Using an RNAi knockdown approach, we show that class A and B J-proteins cooperate to form an interactive flexible network that relocalizes to protein aggregates upon heat shock and preferentially recruits constitutive Hsc70 to disaggregate heat-induced protein aggregates and polyQ aggregates that form in an age-dependent manner. Cooperation between class A and B J-proteins is also required for organismal health and promotes thermotolerance, maintenance of fecundity, and extended viability after heat stress. This disaggregase function of J-proteins and Hsc70 therefore constitutes a powerful regulatory network that is key to Hsc70-based protein quality control mechanisms in metazoa with a central role in the clearance of aggregates, stress recovery, and organismal fitness in aging.


The relationship between socio-demographic factors and depression: a cross sectional study among civilian participants in hostilities in Ukraine.
There is still a gap in scientific knowledge in relation to civilian participants in hostilities. This is despite the fact that there is an extensive body of literature on major depressive disorder (MDD) in individuals who have experienced armed conflict. The purpose of this article is to identify socio-demographic factors which are related to levels of depression among civilian participants in the war in Ukraine, based on a cross-sectional study that was conducted in 2019 from a convenience sample of 314 Ukrainian adults (235 males). Depression was assessed via the Beck Depression Inventory. Multiple regression analyses were conducted to identify possible predictors of depression. Significant predictors were: loss of a loved one, place of residence, age, health insurance, financial situation, and marital status (F (6, 224) = 10.515, p < 0.001, R2 = 0.21; Adjusted R2 = 0.19). They also show that symptoms of depression resulting from the loss of a loved one due to war can be reduced through participation in an educational system. Having children is associated with a risk of more severe depression. Specialists are encouraged to engage in face-to-face interviews and to maintain a supportive and safe environment for participants in hostilities, e.g., in the area of education.


Seroprevalence of Zika virus among asymptomatic pregnant mothers and their newborns in the Najran region of southwest Saudi Arabia.
Zika virus (ZIKV) is a teratogenic flavivirus that can cause microcephaly. Its main vector, Aedes aegypti, has been previ.ously identified in Saudi Arabia, but no ZIKV infection has yet been reported. Nevertheless, the country is at risk from ZIKV because it receives many travelers throughout the year, including pilgrims from ZIKV-endemic countries. Screen asymptomatic pregnant mothers and their newborns attending a major hospital in the Najran region for subclinical or past infections with ZIKV, using ELISA and RT-PCR. Cross-sectional. Najran Maternity and Children Hospital (NMCH). All pregnant women admitted to NMCH in labor between November 2016 and July 2017 were included in the study. Clinical and demographic data were collected by pre-validated physician-administered questionnaires. Paired umbilical and maternal serum samples were collected and frozen at -60°C, using ELISA to measure anti-ZIKA IgG and IgM antibodies and RT-PCR to further investigate positive samples. Maternal and newborn serum anti-ZIKV IgM and IgG and ZIKV RT-PCR. 410 mother-newborn pairs. The median gestational age was 38.5 weeks (range 33-42). Most (n=342, 83.41%) of the women were from Najran city. All of the newborns had normal growth parameters with no congenital malformations. None of the mothers had symptoms suggestive of ZIKV infection; 3 (0.7%) exhibited a low-grade fever (38°C), but did not test positive for anti-ZIKV antibodies. Thirty-five (8.53%) of mothers had travelled inside Saudi Arabia, but none outside the country. Twenty-four (5.85%) mothers tested positive for anti-ZIKV IgM and 52 (12.68%) tested positive for anti-ZIKV IgG, but all infant samples were negative. All seropositive ZIKV IgM were also ZIKV IgG positive, but RT-PCR test.ing of all seropositive samples was negative. Although previous (resolved) ZIKV infection and cross-reactivity of the ELISA method with other flaviviruses cannot be ex.cluded, the study found no confirmed cases of acute ZIKV infection. However, given the presence of the vector in Saudi Arabia, the presence of presumptive positive serology and the ongoing risk of ZIKV entry via a regular influx of travelers from endemic areas, we propose that continuous surveillance be conducted for ZIKV as well for other flaviviruses. Larger-scale nationwide studies are strongly recommended to gain a broader view of the potential threat from ZIKV in the country. Small sample size, unavailability of plaque reduction neutralization tests to confirm serology results, and RT-PCR was only conducted on ELISA-positive serum samples, due to resource constraints. None.


Associations of antiphospholipid antibodies with splanchnic vein thrombosis: a systematic review with meta-analysis.
Splanchnic vein thrombosis (SVT) refers to Budd-Chiari syndrome (BCS) and portal vein system thrombosis (PVST). Current practice guidelines have recommended the routine screening for antiphospholipid antibodies (APAs) in patients with SVT. A systematic review and meta-analysis of observational studies was performed to explore the association between APAs and SVT. The PubMed, EMBASE, and ScienceDirect databases were searched for all relevant papers, in which the prevalence of positive APAs or levels of APAs should be compared between BCS or noncirrhotic PVST patients versus healthy controls, or between cirrhotic patients with portal vein thrombosis (PVT) versus those without PVT. Fourteen studies were eligible. Only 1 study evaluated the role of APAs in BCS patients and found that positive immunoglobulin (Ig) G anticardiolipin antibody (aCL) was more frequently observed in BCS patients than in healthy controls; however, the associations of other APAs with BCS were not evaluated. Positive IgG aCL was more frequently observed in noncirrhotic patients with PVST than in healthy controls; however, other APAs, such as IgM aCL, lupus anticoagulants (LAs), anti-β2-glycoprotein-I antibody (aβ2GPI), and aβ2GPI-oxidized low-density lipoprotein antibody (ox-LDL) were not associated with noncirrhotic PVST. Positive unclassified aCL was more frequently observed in cirrhotic patients with PVT than in those without PVT; however, the association of IgG aCL and IgM aCL with the development of PVT in liver cirrhosis remained inconsistent among studies. The risk of BCS and noncirrhotic PVST might be increased by positive IgG aCL but not IgM aCL, LA, aβ2GPI, or aβ2GPI ox-LDL. However, the evidence regarding APAs in BCS originated from only 1 study. The association between APAs and PVT in liver cirrhosis was unclear.


Techniques of acetabular reconstruction.
Acetabular reconstruction and revision can be a complex and demanding procedure. This article reviews the essential components of preoperative planning and surgical technique. In addition, there are several unsolved issues that will be addressed along the author's current prejudice on these issues.


Cardiac state diagnosis using higher order spectra of heart rate variability.
Heart rate variability (HRV) refers to the regulation of the sinoatrial node, the natural pacemaker of the heart, by the sympathetic and parasympathetic branches of the autonomic nervous system. Heart rate variability analysis is an important tool to observe the heart's ability to respond to normal regulatory impulses that affect its rhythm. A computer-based intelligent system for analysis of cardiac states is very useful in diagnostics and disease management. Like many bio-signals, HRV signals are nonlinear in nature. Higher order spectral analysis (HOS) is known to be a good tool for the analysis of nonlinear systems and provides good noise immunity. In this work, we studied the HOS of the HRV signals of normal heartbeat and seven classes of arrhythmia. We present some general characteristics for each of these classes of HRV signals in the bispectrum and bicoherence plots. We also extracted features from the HOS and performed an analysis of variance (ANOVA) test. The results are very promising for cardiac arrhythmia classification with a number of features yielding a p-value < 0.02 in the ANOVA test.


The Association Between Illness Acceptance and Quality of Life in Women with Breast Cancer.
Breast cancer is the most common cause of cancer death in women. The aim of the study was to investigate the association between illness acceptance and quality of life (QoL) in patients with breast cancer. The study included 150 patients who had undergone surgery for breast cancer. The following standardized questionnaires were used: the Acceptance of Illness Scale (AIS), the EORT QLQ-C30 (The European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire Core 30), and the EORT QL-BR 23 (Quality of Life Questionnaire for Breast Cancer) for QoL evaluation. Socio-clinical data were obtained from the patients' medical records. In the study group, the overall QoL score was 62.67±17.11 in the BCT group, 63±14.3 in the MTX group, and the highest: 65.5±20.2 in the reconstruction group. Comparative analysis showed that patients in the BCT group reported significantly more fatigue (p=0.007) and appetite loss (p=0.032) than those in the MTX+R group. Patients in the MTX group were significantly less satisfied with their body image (p=0.001) and experienced more financial troubles (p=0.013) than the remaining patients. Patients in the MTX+R group reported significantly better sexual function and more sexual enjoyment than the remaining patients (p<0.001). All patients scored high for illness acceptance, though patients in the MTX group had lower scores (28.17±7.2) than the others: 31.84±6.51 in the BCT group and 32.78±7.97 in the MTX+R group. The comparative analysis of QoL according to the level of AIS showed the significantly better QoL and less intense symptoms within all the domains except for the insomnia and hair loss domains in the group of high AIS in comparison with medium and lack of AIS. Acceptance of illness significantly correlated with 4 domains of the QLQ-C30 (p<0.05). The correlation between illness acceptance and overall QoL was positive (r=0.243; p=0.003) - the higher the acceptance, the better the QoL. Correlations with pain, diarrhoea, and financial difficulties were negative. Illness acceptance was positively correlated with QoL in 3 domains of the EORTC-BR23: body image (p<0.001), sexual function (p=0.015), and sexual enjoyment (p=0.047), and negatively with the "treatment side effects" (p=0.011). The level of illness acceptance varies depending on the treatment method, and is the lowest in the group of women having undergone a mastectomy, and the highest in patients after a mastectomy with immediate breast reconstruction. Acceptance of illness improves the QoL of women treated for breast cancer, regardless of the specific treatment method.


NUMERICAL METHODS FOR SOLVING THE MULTI-TERM TIME-FRACTIONAL WAVE-DIFFUSION EQUATION.
In this paper, the multi-term time-fractional wave-diffusion equations are considered. The multi-term time fractional derivatives are defined in the Caputo sense, whose orders belong to the intervals [0,1], [1,2), [0,2), [0,3), [2,3) and [2,4), respectively. Some computationally effective numerical methods are proposed for simulating the multi-term time-fractional wave-diffusion equations. The numerical results demonstrate the effectiveness of theoretical analysis. These methods and techniques can also be extended to other kinds of the multi-term fractional time-space models with fractional Laplacian.


Low extraversion and high neuroticism as indices of genetic and environmental risk for social phobia, agoraphobia, and animal phobia.
The authors examined the extent to which two major personality dimensions (extraversion and neuroticism) index the genetic and environmental risk for three phobias (social phobia, agoraphobia, and animal phobia) in twins ascertained from a large, population-based registry. Lifetime phobias and personality traits were assessed through diagnostic interview and self-report questionnaire, respectively, in 7,800 twins from female-female, male-male, and opposite-sex pairs. Sex-limited trivariate Cholesky structural equation models were used to decompose the correlations among extraversion, neuroticism, and each phobia. In the best-fitting models, genetic correlations were moderate and negative between extraversion and both social phobia and agoraphobia, and that between extraversion and animal phobia was effectively zero. Genetic correlations were high and positive between neuroticism and both social phobia and agoraphobia, and that between neuroticism and animal phobia was moderate. All of the genetic risk factors for social phobia and agoraphobia were shared with those that influence extraversion and neuroticism; in contrast, only a small proportion of the genetic risk factors for animal phobia (16%) was shared with those that influence personality. Shared environmental experiences were not a source of correlations between personality traits and phobias, and unique environmental correlations were relatively modest. Genetic factors that influence individual variation in extraversion and neuroticism appear to account entirely for the genetic liability to social phobia and agoraphobia, but not animal phobia. These findings underline the importance of both introversion (low extraversion) and neuroticism in some psychiatric disorders.


scruff: an R/Bioconductor package for preprocessing single-cell RNA-sequencing data.
Single-cell RNA sequencing (scRNA-seq) enables the high-throughput quantification of transcriptional profiles in single cells. In contrast to bulk RNA-seq, additional preprocessing steps such as cell barcode identification or unique molecular identifier (UMI) deconvolution are necessary for preprocessing of data from single cell protocols. R packages that can easily preprocess data and rapidly visualize quality metrics and read alignments for individual cells across multiple samples or runs are still lacking. Here we present scruff, an R/Bioconductor package that preprocesses data generated from the CEL-Seq or CEL-Seq2 protocols and reports comprehensive data quality metrics and visualizations. scruff rapidly demultiplexes, aligns, and counts the reads mapped to genome features with deduplication of unique molecular identifier (UMI) tags. scruff also provides novel and extensive functions to visualize both pre- and post-alignment data quality metrics for cells from multiple experiments. Detailed read alignments with corresponding UMI information can be visualized at specific genome coordinates to display differences in isoform usage. The package also supports the visualization of quality metrics for sequence alignment files for multiple experiments generated by Cell Ranger from 10X Genomics. scruff is available as a free and open-source R/Bioconductor package. scruff streamlines the preprocessing of scRNA-seq data in a few simple R commands. It performs data demultiplexing, alignment, counting, quality report and visualization systematically and comprehensively, ensuring reproducible and reliable analysis of scRNA-seq data.


User Perceptions and Experiences of a Handheld 12-Lead Electrocardiographic Device in a Clinical Setting: Usability Evaluation.
Cardiac arrhythmias are a leading cause of death. The mainstay method for diagnosing arrhythmias (eg, atrial fibrillation) and cardiac conduction disorders (eg, prolonged corrected QT interval [QTc]) is by using 12-lead electrocardiography (ECG). Handheld 12-lead ECG devices are emerging in the market. In tandem with emerging technology options, evaluations of device usability should go beyond validation of the device in a controlled laboratory setting and assess user perceptions and experiences, which are crucial for successful implementation in clinical practice. This study aimed to evaluate clinician and patient perceptions and experiences, regarding the usability of a handheld 12-lead ECG device compared to a conventional 12-lead ECG machine, and generalizability of this user-centered approach. International Organization for Standardization Guidelines on Usability and the Technology Acceptance Model were integrated to form the framework for this study, which was conducted in outpatient clinics and cardiology wards at Westmead Hospital, New South Wales, Australia. Each patient underwent 2 ECGs (1 by each device) in 2 postures (supine and standing) acquired in random sequence. The times taken by clinicians to acquire the first ECG (efficiency) using the devices were analyzed using linear regression. Electrocardiographic parameters (QT interval, QTc interval, heart rate, PR interval, QRS interval) and participant satisfaction surveys were collected. Device reliability was assessed by evaluating the mean difference of QTc measurements within ±15 ms, intraclass correlation coefficient, and level of agreement of the devices in detecting atrial fibrillation and prolonged QTc. Clinicians' perceptions and feedback were assessed with semistructured interviews based on the Technology Acceptance Model. A total of 100 patients (age: mean 57.9 years, SD 15.2; sex: male: n=64, female n=36) and 11 clinicians (experience acquiring ECGs daily or weekly 10/11, 91%) participated, and 783 ECGs were acquired. Mean differences in QTc measurements of both handheld and conventional devices were within ±15 ms with high intraclass correlation coefficients (range 0.90-0.96), and the devices had a good level of agreement in diagnosing atrial fibrillation and prolonged QTc (κ=0.68-0.93). Regardless of device, QTc measurements when patients were standing were longer duration than QTc measurements when patients were supine. Clinicians' ECG acquisition times improved with usage (P<.001). Clinicians reported that device characteristics (small size, light weight, portability, and wireless ECG transmission) were highly desired features. Most clinicians agreed that the handheld device could be used for clinician-led mass screening with enhancement in efficiency by increasing user training. Regardless of device, patients reported that they felt comfortable when they were connected to the ECG devices. Reliability and usability of the handheld 12-lead ECG device were comparable to those of a conventional ECG machine. The user-centered evaluation approach helped us identify remediable action to improve the efficiency in using the device and identified highly desirable device features that could potentially help mass screening and remote assessment of patients. The approach could be applied to evaluate and better understand the acceptability and usability of new medical devices.


Assessing the quality of dental services using SERVQUAL model.
The measurement of service quality had an important role in managing service provided, diagnosing the problem, and assessing service performance. Patient satisfaction is one of the key indicators of quality in health-care organizations. This study aimed to measure the service quality gap of patients' perceptions and expectations in five dimensions at a military Specialized Dental Clinic in Iran using SERVQUAL tool. This was a cross-sectional and descriptive-analytical study conducted at a Specialized Dental Clinic affiliated to the Armed Forces of Islamic Republic of Iran in Tehran in 2013. All 385 patients referring to the studied clinic during two working shifts of morning and evening in one month, August, were selected. The required data were collected using the modified SERVQUAL questionnaire. The collected data were analyzed using SPSS 20.0 through statistical tests including Mann-Whitney and Kruskal-Wallis tests. P < 0.05 was considered statistically significant. The results showed that the quality of services provided to patients was significantly lower than their expectations and the quality gaps were statistically significant in all studied dimensions (P < 0.001). Furthermore, the highest and lowest quality gap was related to empathy (-1.16) and reliability (-0.61), respectively. According to the results, the managers of this clinic should take steps toward improving the quality of services in all dimensions, especially responsiveness and empathy, through planning properly, prioritizing services, and reviewing processes with regard to the patients' expectations.


Regulation of human nucleus pulposus cells by peptide-coupled substrates.
Nucleus pulposus (NP) cells are derived from the notochord and differ from neighboring cells of the intervertebral disc in phenotypic marker expression and morphology. Adult human NP cells lose this phenotype and morphology with age in a pattern that contributes to progressive disc degeneration and pathology. Select laminin-mimetic peptide ligands and substrate stiffnesses were examined for their ability to regulate human NP cell phenotype and biosynthesis through the expression of NP-specific markers aggrecan, N-cadherin, collagen types I and II, and GLUT1. Peptide-conjugated substrates demonstrated an ability to promote expression of healthy NP-specific markers, as well as increased biosynthetic activity. We show an ability to re-express markers of the juvenile NP cell and morphology through control of peptide presentation and stiffness on well-characterized polyacrylamide substrates. NP cells cultured on surfaces conjugated with α3 integrin receptor peptides P4 and P678, and on α2, α5, α6, β1 integrin-recognizing peptide AG10, show increased expression of aggrecan, N-cadherin, and types I and II collagen, suggesting a healthier, more juvenile-like phenotype. Multi-cell cluster formation was also observed to be more prominent on peptide-conjugated substrates. These findings indicate a critical role for cell-matrix interactions with specific ECM-mimetic peptides in supporting and maintaining a healthy NP cell phenotype and bioactivity. NP cells reside in a laminin-rich environment that deteriorates with age, including a loss of water content and changes in the extracellular matrix (ECM) structure that may lead to the development of a degenerated IVD. There is great interest in methods to re-express healthy, biosynthetically active NP cells using laminin-derived biomimetic peptides toward the goal of using autologous cell sources for tissue regeneration. Here, we describe a novel study utilizing several laminin mimetic peptides conjugated to polyacrylamide gels that are able to support an immature, healthy NP phenotype after culture on "soft" peptide gels. These findings can support future studies in tissue regeneration where cells may be directed to a desired regenerative phenotype using niche-specific ECM peptides.


Complex interventions can increase osteoporosis investigations and treatment: a systematic review and meta-analysis.
Osteoporosis is affecting over 200 million people worldwide. Despite available guidelines, care for these patients remains sub-optimal. We developed an osteoporosis tool to address the multiple dimensions of chronic disease management. Findings from its evaluation showed a significant increase from baseline in osteoporosis investigations and treatment, so we are revising this tool to include multiple chronic conditions including an update of evidence about osteoporosis. Our objectives were to conduct a systematic review of osteoporosis interventions in adults at risk for osteoporosis. We searched bibliometric databases for randomized controlled trials (RCTs) in any language evaluating osteoporosis disease management interventions in adults at risk for osteoporosis. Reviewer pairs independently screened citations and full-text articles, extracted data, and assessed risk of bias. Analysis included random effects meta-analysis. Primary outcomes were osteoporosis investigations and treatment, and fragility fractures. Fifty-five RCTs and one companion report were included in the analysis representing 165,703 patients. Our findings from 55 RCTs and 18 sub-group meta-analyses showed that complex implementation interventions with multiple components consisting of at least education + feedback + follow-up significantly increased the initiation of osteoporosis medications, and interventions with at least education + follow-up significantly increased the initiation of osteoporosis investigations. No significant impact was found for any type of intervention to reduce fracture. Complex interventions that include at least education + follow-up or feedback have the most potential for increasing osteoporosis investigations and treatment. Patient education appears to be an important component in osteoporosis disease management.


An increased platelet-leukocytes interaction at the culprit site of coronary artery occlusion in acute myocardial infarction: a pathogenic role for "no-reflow" phenomenon?
Distal protection devices have been shown to reduce the incidence of "no flow" phenomenon during primary percutaneous coronary intervention (PCI). So far, it has not been well clarified which mechanism is mainly involved in distal coronary protection. To investigate the activation state of leukocytes and platelets locally present within the blood from the site of coronary occlusion. Ten patients with acute myocardial infarction (AMI) underwent primary PCI with an embolization protection device and aspiration catheter (PercuSurge GuardWire) were included. The following functional parameters: a) monocyte and neutrophils surface molecules; b) platelet surface activatory antigens; c) leukocytes-platelet conjugates were studied by flow cytometry in blood obtained from the site of coronary occlusion and from peripheral femoral artery. The leukocyte-platelet adhesion index was significantly higher in the aspirated blood at the site of coronary occlusion than in the peripheral arterial blood for both monocytes (0.226+/-0.04 vs. 0.084+/-0.01; p=0.004) and neutrophils (1.372+/-0.3 vs. 0.524+/-0.1; p=0.02). Moreover, the volume of coaggregates exhibited a significant increase in coronary blood for both populations (p=0.02 for monocytes and for neutrophils). Interestingly, a significant up-regulation of the adhesive molecule CD18 was observed in coronary blood respect to systemic circulation either in monocytes (p=0.01) than in neutrophils (p=0.003). A significant up-regulation of monocyte (HLA-DR) and neutrophil (CD66b) activatory molecules expression was also observed in the aspirated coronary compared to peripheral artery blood (p=0.02 and p=0.03 for HLA-DR and CD66b, respectively). These data indicate an increased leukocyte-platelet functional interaction in AMI at the site of plaque rupture relative to the systemic circulation, which may be one of the pathogenetic mechanisms responsible for myocardial "no-reflow" phenomenon.


Patients' perspective, quality of life and treatment goals in Hailey-Hailey disease: Lessons learned from the German National Registry.
Hailey-Hailey disease (HHD) remains a difficult-to-treat dermatosis and little is known about the patient's perception of the disease activity, the treatment success and its impact on quality-of-life (QoL). To obtain better understanding of HHD patients' needs regarding their medical condition, financial burden, QoL, subjective well-being and treatment thereof as well as satisfaction to evaluate common treatments' 'real-life' relevance. With initiation of the national registry for Darier's disease (DD; Morbus Darier, MD) and Hailey-Hailey disease (HH) MDHHgermany, patients with HHD diagnosis were included starting June 2020. To assess subjective symptoms, patients filled out questionnaires such as the DLQI (dermatological life quality index), numeric rating scale (NRS) for itch, pain and burning sensation, as well as the SWLS (satisfaction with life scale) questionnaire to quantify overall satisfaction in life. Additionally, data on therapies were collected along with the patients' satisfaction of those and their medical care. Furthermore, patients assessed financial aspects and work ability. One hundred and two patients were recruited from dermatology clinics, office-based dermatologists and self-help platforms across Germany between June 2020 and February 2023, 90 were eligible and analysed (mean: 49.91 years, 73.33% females, 26.67% males). 39.77% stated according to the DLQI their life is severely/very severely affected. Satisfaction with life was mediocre. Burning sensation was most pronounced among subjective symptoms (NRS 5.85 ± 2.80). Systemic treatments were rated as ineffective according to 56.92%, 25.56% had never received one. Most prescribed systemic treatments were corticosteroids (73.8%), followed by low-dose naltrexone (LDN) (26.2%), retinoids (15.4%) and antibiotics (13.8%). Satisfaction with medical care was generally low. Our 'real-life' data state a major disease burden and impact on the QoL for affected individuals, as well as limited disease control due to inadequate therapies. MDHHgermany can provide insights into improvement of healthcare support with this debilitating disease and improve QoL. In the long term, it aims to provide basis for further clinical trials, epidemiological studies and immunological investigations.


Structure of supported DPPC/cholesterol bilayers studied via X-ray reflectivity.
The electron density profile of bilayers of DPPC/cholesterol mixtures supported on semiconductor grade silicon substrates were studied with the objective of determining how the proximity of a solid interface modifies the phase diagram of mixed bilayers. The bilayers were studied in situ immersed in water via synchrotron X-ray reflectivity (XRR). Measurements were performed as a function of temperature through the main phase transition and cholesterol mole fractions up to 40%. Analysis of XRR yields the bilayer thickness, roughness and leaflet asymmetry. We find that the structure of the pure DPPC bilayers in the gel phase is in agreement with previous X-ray measurements of supported bilayers deposited via vesicle fusion and multilamellar vesicles but show more clearly defined features than measurements made on films formed using Langmuir-Blodget Langmuir-Shaffer (LB) deposition. Examination of bilayer thickness vs. temperature shows that the melting temperature for supported bilayers is shifted upwards by approximately 4 °C relative to multilamellar vesicles and that the melting temperature decreases with increasing cholesterol content up to 20%. For pure DPPC bilayers the leaflets melt in two stages with the distal leaflet melting first. For cholesterol concentrations of 10% and 20% there is no clear indication of separate melting. For 33% and 40% cholesterol content no clear transition is seen in the bilayer thickness, but an abrupt change in roughness indicates possible microdomain formation in the 40% cholesterol sample.


The effects of acute and chronic ethanol administration and its withdrawal on gamma-aminobutyric acid receptor binding in rat brain.
1 The effects of acute and chronic ethanol administration, and withdrawal on the binding of the inhibitory neurotransmitter, gamma-aminobutyric acid (GABA), was investigated in rat brain. 2 Acute ethanol (2 to 4 g/kg i.p. 30 min before removal of brain) produced an increase in the binding capacity of the low affinity GABA receptor binding site. 3 Following chronic ethanol administration (1 to 21 days), the GABA receptor binding characteristics were not altered. These results suggest a possible adaptation of GABA receptors to the continuous presence of ethanol at the GABA synapse. 4 During ethanol withdrawal, the affinity of the low affinity GABA receptor binding site was significantly lower than pair-fed controls at 8 and 16 h withdrawal. 5 These results suggest that GABA receptor sensitivity may play a role in some of the neuropharmacological effects of ethanol and in its withdrawal symptoms.


Cu(ii), Ga(iii) and In(iii) complexes of 2-acetylpyridine N(4)-phenylthiosemicarbazone: synthesis, spectral characterization and biological activities.
In this paper, synthesis and characterization of metal complexes [Cu2(L)3]ClO4 (1), [Ga(L)2]NO3·2H2O (2) and [In(L)2]NO3·H2O (3) (HL = 2-acetylpyridine N(4)-phenylthiosemicarbazone) was carried out, including elemental analysis, spectral analysis (IR, UV-vis, NMR), and X-ray crystallography. Complex 1 contains one S-bridged binuclear [Cu2(L)3]+ unit, where two Cu atoms display diverse coordination geometries: one being square planar geometry and the other octahedral geometry. Both 2 and 3 are mononuclear complexes, and the metal centers in 2 and 3 are chelated by two NNS tridentate ligands possessing a distorted octahedral geometry. Biological studies show that all the complexes possess a wide spectrum of modest to effective antibacterial activities and remarkable cytotoxicities against HepG2 cells, and 1, in particular, with an IC50 value of 0.19 ± 0.06 μM, is 113-fold and 28-fold more cytotoxic than HL and the antitumor drug mitoxantrone, respectively. In addition, 3 exhibits excellent photoluminescence properties. Upon the addition of 1 equiv of In3+ ions, a remarkable fluorescence intensity of HL and fluorescent color change (from transparent to light-green) could be observed with 365 nm light, indicating that this ligand may be used as a promising colorimetric and fluorescent probe for In3+ detection.


Monoglycerides and fatty acids from Ibervillea sonorae root: isolation and hypoglycemic activity.
Eleven monoglycerides (MG), 1-monopalmitin (1), glyceryl 1-monomargarate (2), 1-monostearin (3), glyceryl 1-monononadecylate ( 4), glyceryl 1-monoarachidate (5), glyceryl 1-monobehenate (6), glyceryl 1-monotricosanoate (7), glyceryl 1-monotetracosanoate (8), glyceryl 1-monopentacosanoate (9), glyceryl 1-monohexacosanoate (10) and glyceryl 1-monooctacosanoate (11), together with five fatty acids (FA), lauric acid (12), myristic acid (13), pentadecanoic acid (14), palmitic acid (15) and stearic acid (16) were isolated of the root of IBERVILLEA SONORAE Greene (Cucurbitaceae). Their structures were determined by spectroscopic and chemical methods as well as GC-MS analysis. The hypoglycemic activity of the dichloromethane (DCM) extract, of fractions (F1-F10 and SF1-SF5), of monoglycerides (MG) and of fatty acids (FA) mixtures obtained of the root from I. SONORAE was evaluated in normoglycemic and alloxan-induced diabetic mice. The results showed that by intraperitoneal administration the DCM extract (300 mg/kg), F9 (300 mg/kg) and SF1 (150 mg/kg) significantly reduced glucose levels in both models. For fraction SF1, the hypoglycemic activity was more pronounced than that of tolbutamide (150 mg/kg) used as control. However, neither MG (75 mg/kg) nor FA (75 mg/kg) mixtures isolated from SF1 exhibited a significant hypoglycemic effect. However, when MG and FA were combined in equal proportions (75 mg: 75 mg/kg), their effect was comparable to that of SF1. The observed activity for the DCM extract, F9, SF1 and the MG-FA mixture provides additional support for the popular use of this plant in the treatment of diabetes mellitus in Mexican traditional medicine.


Selected wheat seed defense proteins exhibit competitive binding to model microbial lipid interfaces.
Puroindolines (Pins) and purothionins (Pths) are basic, amphiphilic, cysteine-rich wheat proteins that play a role in plant defense against microbial pathogens. This study examined the co-adsorption and sequential addition of Pins (Pin-a, Pin-b, and a mutant form of Pin-b with Trp-44 to Arg-44 substitution) and β-purothionin (β-Pth) model anionic lipid layers using a combination of surface pressure measurements, external reflection FTIR spectroscopy, and neutron reflectometry. Results highlighted differences in the protein binding mechanisms and in the competitive binding and penetration of lipid layers between respective Pins and β-Pth. Pin-a formed a blanket-like layer of protein below the lipid surface that resulted in the reduction or inhibition of β-Pth penetration of the lipid layer. Wild-type Pin-b participated in co-operative binding with β-Pth, whereas the mutant Pin-b did not bind to the lipid layer in the presence of β-Pth. The results provide further insight into the role of hydrophobic and cationic amino acid residues in antimicrobial activity.


Tears of the peripheral retina; pathogenesis, incidence and classification in autopsy eyes.
A pathogenic analysis of full-thickness tears of the peripheral retina and their incidence in autopsy eyes is presented. Tears were classified according to their location (ora or postora, quadrant and zone), type of traction (zonuloretinal or vitreoretinal), morphological features (flap or operculum), presence or absence of posterior vitreous detachment, and their association with other ocular lesions. Tears were present in 2% of eyes. They were postoral in 92%, uniformly distributed in the quadrants, in the equatorial zone in 95%, related to vitreoretinal traction in 94%, of the flap type in 64%, accompanied by posterior vitreous detachment in 79% and were significantly more prevalent in eyes with lattice degeneration (17%). A simple classification of postoral tears is recommended which incorporates these pathogenic principles and includes three types according to their topographical relationship to the vitreous base: intrabasal--caused by avulsion of zonular traction tufts; juxtabasal--related to traction of the posteriorly detached vitreous on irregularities in posterior border of vitreous base; extrabasal--resulting from avulsion of cystic retinal tufts.


Augmenting occupational therapy treatment of upper-extremity spasticity with botulinum toxin A: a case report of progress at discharge and 2 years later.
This case report summarizes occupational therapy services provided to a participant after botulinum toxin A (BTA) injections and her improvements 2 years later. The injections were performed to neutralize spasticity in the left upper extremity after a stroke. Along with BTA, the participant received occupational therapy for 12 weeks, using neurodevelopmental and biomechanical approaches and an activity-based home program. Spasticity decreased from a 4 to a 1 on the Modified Ashworth Scale (MAS) immediately after the injections. Two years after the injections, the MAS score ranged between 1 and 1+. Initially, the participant demonstrated functional limitations in areas of splint application, dressing, toileting, and bathing. Following BTA injections and occupational therapy, the participant demonstrated increased independence in all deficit areas. For this participant, BTA, combined with functional, activity-based occupational therapy interventions, was associated with neurological change and greater functional use of the spastic limb.


Recombinant high-density lipoprotein complex as a targeting system of nosiheptide to liver cells.
Nosiheptide is a lipophilic peptide of significant anti-hepatitis B virus (anti-HBV) activity in cell culture, but has poor distribution to liver in vivo. In this study, recombinant high-density lipoprotein (rHDL) complexes of nosiheptide were constructed to target this anti-HBV agent to hepatocytes. The optimized rHDL-nosiheptide complex had a high drug-loading efficiency (>80%) and a diameter smaller than 30 nm. The concentration of nosiheptide in an optimized rHDL-nosiheptide complex to achieve 50% virus inhibition (IC(50)) in HepG(2) 2.2.15 cells was 0.63 microg/ml, which was 40 times lower than the IC(50) of nosiheptide in control liposome (2.5 microg/ml) and 200 times lower than the IC(50) of the free nosiheptide (12.5 microg/ml). The complex targeted most of the administered nosiheptide to the liver within 30 min after i.v. injection to male Wistar rats. Together, this report provides early evidence that it is feasible to develop efficient, HDL-based drug delivery systems against HBV, utilizing apolipoprotein A-I as the targeting moiety.


Quantum Chemical Methods for Modeling Covalent Modification of Biological Thiols.
Targeted covalent inhibitor drugs require computational methods that go beyond simple molecular-mechanical force fields in order to model the chemical reactions that occur when they bind to their targets. Here, several semiempirical and density-functional theory (DFT) methods are assessed for their ability to describe the potential energy surface and reaction energies of the covalent modification of a thiol by an electrophile. Functionals such as PBE and B3LYP fail to predict a stable enolate intermediate. This is largely due to delocalization error, which spuriously stabilizes the prereaction complex, in which excess electron density is transferred from the thiolate to the electrophile. Functionals with a high-exact exchange component, range-separated DFT functionals, and variationally optimized exact exchange (i.e., the LC-B05minV functional) correct this issue to various degrees. The large gradient behavior of the exchange enhancement factor is also found to significantly affect the results, leading to the improved performance of PBE0. While ωB97X-D and M06-2X were reasonably accurate, no method provided quantitative accuracy for all three electrophiles, making this a very strenuous test of functional performance. Additionally, one drawback of M06-2X was that molecular dynamics (MD) simulations using this functional were only stable if a fine integration grid was used. The low-cost semiempirical methods, PM3, AM1, and PM7, provide a qualitatively correct description of the reaction mechanism, although the energetics is not quantitatively reliable. As a proof of concept, the potential of mean force for the addition of methylthiolate to methylvinyl ketone was calculated using quantum mechanical/molecular mechanical MD in an explicit polarizable aqueous solvent. © 2019 Wiley Periodicals, Inc.


Non-pharmacological interventions for improving sleep quality in patients on dialysis: systematic review and meta-analysis.
We conducted a meta-analysis to summarise and quantify the effects of non-pharmacological interventions on sleep quality improvement in uraemic patients on dialysis. We defined the primary outcome as the change of sleep quality before and after interventions (evaluated by polysomonography or subjective questionnaires such as Pittsburgh sleep quality index, PSQI). The change of fatigue scales, inflammatory cytokines and adverse events were analysed as secondary outcomes. Twelve eligible randomised controlled trials and one prospective cohort study were identified. All three identified non-pharmacological interventions could result in a greater PSQI score reduction compared to controls: 1) cognitive-behavioural therapy (CBT) versus sleep hygiene education (standardised mean difference (SMD) 0.85, 95% CI 0.37-1.34); 2) physical training versus no training (SMD 3.36, 95% CI 2.16-4.57) and 3) Acupressure (including other acupoints massages) versus control (SMD 1.77, 95% CI 0.80-2.73). In terms of subscores, we found that CBT may shorten sleep latency, alleviate sleep disturbance and reduce the use of sleep medications. The finding of the cohort study suggested that intradialytic aerobic exercise training improved sleep quality in haemodialysis patients with restless leg syndrome. In conclusion, in dialysis-dependent patients, CBT could shorten sleep latency, alleviate sleep disturbance and reduce the use of sleep medications. Acupressure (including other acupoints massages) and exercise training are promising interventions but the results in these subgroups should be interpreted cautiously due to the concern of methodological quality and potential confounding factors.


Identification and subtype analysis of biomarkers associated with the solute carrier family in acute myocardial infarction.
The dysregulation of some solute carrier (SLC) proteins has been linked to a variety of diseases, including diabetes and chronic kidney disease. However, SLC-related genes (SLCs) has not been extensively studied in acute myocardial infarction (AMI). The GSE66360 and GSE60993 datasets, and SLCs geneset were enrolled in this study. Differentially expressed SLCs (DE-SLCs) were screened by overlapping DEGs between the AMI and control groups and SLCs. Next, functional enrichment analysis was carried out to research the function of DE-SLCs. Consistent clustering of samples from the GSE66360 dataset was accomplished based on DE-SLCs selected. Next, the gene set enrichment analysis (GSEA) was performed on the DEGs-cluster (cluster 1 vs cluster 2). Three machine learning models were performed to obtain key genes. Subsequently, biomarkers were obtained through receiver operating characteristic (ROC) curves and expression analysis. Then, the immune infiltration analysis was performed. Afterwards, single-gene GSEA was carried out, and the biomarker-drug network was established. Finally, quantitative real-time fluorescence PCR (qRT-PCR) was performed to verify the expression levels of biomarkers. In this study, 13 DE-SLCs were filtered by overlapping 366 SLCs and 448 DEGs. The functional enrichment results indicated that the genes were implicated with amino acid transport and TNF signaling pathway. After the consistency clustering analysis, the samples were classified into cluster 1 and cluster 2 subtypes. The functional enrichment results showed that DEGs-cluster were implicated with chemokine signaling pathway and so on. Further, SLC11A1 and SLC2A3 were identified as SLC-related biomarkers, which had the strongest negative relationship with resting memory CD4 T cells and the strongest positive association with activated mast cells. In addition, the single-gene GSEA results showed that cytosolic ribosome was enriched by the biomarkers. Five drugs targeting SLC2A3 were predicted as well. Lastly, the experimental results showed that the biomarkers expression trends were consistent with public database. In this study, 2 SLC-related biomarkers (SLC11A1 and SLC2A3) were screened and drug predictions were carried out to explore the prediction and treatment of AMI.


Short-term Glycemic Variability and Its Association With Macrovascular and Microvascular Complications in Patients With Diabetes.
The introduction of continuous glucose monitoring inaugurated a new era in clinical practice by shifting the characterization of glycemic control from HbA1c to novel metrics. The one that gained widespread attention over the past decades was glycemic variability (GV), which typically refers to peaks and nadirs of blood glucose measured over a given time interval. GV can be dichotomized into two main categories: short-term and long-term. Short-term GV reflects within-day and between-day glycemic oscillations, and its contribution to diabetic complications remains an enigma. In this review, we summarize the available data about short-term GV and its possible association with both microvascular and macrovascular complications, evaluating different pathogenic mechanisms and demonstrating nonpharmaceutical, as well as pharmaceutical, therapeutic interventions.


The human immunodeficiency virus (HIV) type 2 envelope protein is a functional complement to HIV type 1 Vpu that enhances particle release of heterologous retroviruses.
We have recently shown that the envelope glycoprotein of the ROD10 isolate of human immunodeficiency virus type 2 (HIV-2) has the ability to positively regulate HIV-2 viral particle release. The activity provided by the ROD10 Env was remarkably similar to that of the HIV-1 Vpu protein, thus raising the possibility that the two proteins act in a related fashion. We now show that the ROD10 Env can functionally replace Vpu to enhance the rate of HIV-1 particle release. When provided in trans, both Vpu and the ROD10 Env restored wild-type levels of particle release in a Vpu-deficient mutant of the NL4-3 molecular clone with indistinguishable efficiencies. This effect was independent of the presence of the HIV-1 envelope protein. The ROD10 Env also enhanced HIV-1 particle release in the context of HIV-2 chimeric viruses containing the HIV-1 gag-pol, indicating a lack of need for additional HIV-1 products in this process. In addition, we show for the first time that HIV-1 Vpu, as well as ROD10 Env, has the ability to enhance simian immunodeficiency virus (SIV) particle release. The effects of Vpu and ROD10 Env on SIV particle release were indistinguishable and were observed in the context of full-length SIVmac239 and simian-human immunodeficiency virus chimeras. These results further demonstrate that ROD10 Env can functionally complement Vpu with respect to virus release. In contrast, we found no evidence of a destabilizing activity of ROD10 Env on the CD4 molecule. HIV-1 and HIV-2 thus appear to have evolved genetically distinct but functionally similar strategies to resolve the common problem of efficient release of progeny virus from infected cells.


Determination of Clorsulon and its related substances in commercial bulk drug substance batches by a rapid ion-pair UPLC method.
Clorsulon is a potent anthelmintic agent and is widely used for the treatment and control of parasites including adult liver flukes in cattle and sheep. A rapid ion-paired reversed phase ultraperformance liquid chromatography (IP-UPLC) method has been developed and validated for determination of Clorsulon and its related substances in bulk drug substance batches of Clorsulon with a short octadecyl column. Analytes were eluted by a gradient elution on a Acquity UPLC® BEH C18 column (50 mm × 2.1 mm i.d., 1.7 µm particle size). Column temperature was maintained at 55 °C and all analytes were monitored by UV detection at 268 nm. Mobile phase-A constitutes 3 mM tetrabutylammonium hydroxide in H2O and mobile phase-B constitutes acetonitrile/methanol (50/50, v/v), respectively. All analytes of interest were adequately separated within 5 min. The analytes were quantitated against an external reference standard of Clorsulon. The chemical structures of degradation products of Clorsulon were proposed based on two-dimensional UPLC-MS data. The IP-UPLC method has been successfully validated and demonstrated to be accurate, sensitive, robust, and specific.


A 1-megadalton translocation complex containing Tic20 and Tic21 mediates chloroplast protein import at the inner envelope membrane.
Chloroplast protein import is mediated by two hetero-oligomeric protein complexes, the Tic and Toc translocons, which are located in the inner and outer envelope membranes. At the inner membrane, many Tic components have been identified and characterized, but it remains unclear how these Tic proteins are organized to form a protein-conducting channel or whether a stable Tic core complex that binds translocating preproteins exists. Here, we report the identification of a 1-megadalton (MD) translocation complex as an intermediate during protein translocation across the inner membrane in Arabidopsis thaliana and pea (Pisum sativum). This complex can be detected by blue native PAGE using the mild detergent digitonin without any chemical cross-linkers. The preprotein arrested in the 1-MD complex can be chased into its fully translocated form after a subsequent incubation. While Tic20 and Tic21 appear to be involved in the 1-MD complex, Tic110, a well-characterized Tic component, exists as a distinct entity from the complex. Several lines of evidence suggest that the 1-MD complex functions in between the Toc and Tic110-containing complexes, most likely as a protein-conducting channel at the inner envelope.


Characterization of native and oxidized human low-density lipoproteins by the Z-scan technique.
The nonlinear optical response of human normal and oxidized by Cu2+ low-density lipoproteins particles (LDL), were investigated by the Z-scan technique as a function of temperature and concentration of LDL particles. The Z-scan signals increase linearly with concentration of normal LDL particles, following the usual Beer-Lambert law in a broad range of concentrations. The oxidized LDL particles do not show nonlinear optical response. On the other hand, normal LDL increases its nonlinear optical response as a function of temperature. These behaviors can be attributed to an absorbing element that is modified by the oxidative process. Contrarily, changes in the physical state of the cores and conformation of the ApoB100 protein due to an increase in temperature seems to enhance their nonlinear optical properties. This tendency is not due to aggregation of particles. The main contribution to the nonlinear optical response of normal LDL particles comes from the phospholipid fraction of the particles.


Cerebral oxygenation during skin-to-skin care in preterm infants not receiving respiratory support.
Skin-to-skin care (SSC) has proven benefits in preterm infants, but increased hypoxic and bradycardic events have been reported. This may make clinicians hesitant to recommend SSC as standard care. We hypothesised that regional cerebral oxygenation (rStO2) measured with near infrared spectroscopy is not worse during SSC compared with standard incubator care. Prospective, observational, non-inferiority study. Single tertiary perinatal centre in Australia. Forty preterm infants (median (IQR) 30.6 (29.1-31.7) weeks' gestation) not receiving respiratory support were studied on day 14 (8-38). Recordings during 90 min of incubator care, followed by 90 min of SSC. Each infant acted as their own control and caregivers were blinded to the rStO2 measurements. The primary outcome was the mean difference in rStO2 between SSC and incubator care. The prespecified margin of non-inferiority was -1.5%. Secondary outcomes included heart rate (HR), peripheral oxygen saturation (SpO2), time in quiet sleep, temperature and hypoxic (SpO2 <80% for >5 s) or bradycardic events (HR <80 bpm for >5 s) and time spent in cerebral hypoxia (rStO2<55%) and hyperoxia (rStO2>85%). Mean (SD) rStO2 was lower during SSC compared with incubator care: 73.6 (6.0)% vs 74.8 (4.6)%, mean difference (95% CI) 1.3 (2.2 to 0.4)%. HR was 5 bpm higher, SpO2 1% lower and time in quiet sleep 24% longer during SSC. Little evidence of a difference was observed in temperature. The number of hypoxic or bradycardic events as well as the proportion of time spent in cerebral hypoxia and hyperoxia was very low in both periods. Mean rStO2 was marginally lower during SSC without observed differences in hypoxic or bardycardic events but an increase in time spent in quiet sleep. This trial is linked to Australian New Zealand Clinical Trials Registry: identifier 12616000240448. It was registered pre-results.


Ultrastructure of Calymmatobacterium granulomatis: comparison of culture with tissue biopsy specimens.
The ultrastructural features of cells of Calymmatobacterium granulomatis from monocyte co-cultures and tissue biopsy specimens were compared. In cultures the bacteria were mainly extracellular, i.e., not within membrane-bound vacuoles. The bacterial body was surrounded by a uniformly extensive homogeneous layer with a relatively high electron density. This layer varied considerably in tissue biopsy specimens, having either homogeneously electron-dense or delicate web-like structures with varying density and thickness. In tissue specimens the bacteria were located predominantly within vacuoles of varying sizes in the cytoplasm of the macrophages and, occasionally, extracellularly within the intercellular spaces of the stroma. The bacterial cytoplasm contained ribosomes scattered throughout with electron-dense granules located peripherally. The trilaminar cell-wall structure was typical of a gram-negative organism, comprising an outer membrane, a middle electron-opaque layer and an inner plasma membrane. Surface structures such as fimbriae, flagella and bacteriophages were not identified in specimens from either source.


Characterization of the alternative sigma factor sigma54 and the transcriptional regulator FleQ of Legionella pneumophila, which are both involved in the regulation cascade of flagellar gene expression.
We cloned and analyzed Legionella pneumophila Corby homologs of rpoN (encoding sigma(54)) and fleQ (encoding sigma(54) activator protein). Two other genes (fleR and pilR) whose products have a sigma(54) interaction domain were identified in the genome sequence of L. pneumophila. An rpoN mutant strain was nonflagellated and expressed very small amounts of the FlaA (flagellin) protein. Like the rpoN mutant, the fleQ mutant strain of L. pneumophila was also nonflagellated and expressed only small amounts of FlaA protein compared to the amounts expressed by the wild type. In this paper we show that the sigma(54) factor and the FleQ protein are involved in regulation of flagellar gene operons in L. pneumophila. RpoN and FleQ positively regulate the transcription of FliM and FleN, both of which have a sigma(54)-dependent promoter consensus sequence. However, they seemed to be dispensable for transcription of flaA, fliA, or icmR. Our results confirmed a recently described model of the flagellar gene regulation cascade in L. pneumophila (K. Heuner and M. Steinert, Int. J. Med. Microbiol. 293:133-145, 2003). Flagellar gene regulation was found to be different from that of Enterobacteriaceae but seems to be comparable to that described for Pseudomonas or Vibrio spp.


Pentoxifylline does not decrease short-term mortality but does reduce complications in patients with advanced cirrhosis.
Pentoxifylline, an inhibitor of tumor necrosis factor-alpha, is given to patients with liver diseases, but its effects in patients with advanced cirrhosis are unknown. We performed a randomized, placebo-controlled, double-blind trial of its effects in patients with cirrhosis. A total of 335 patients with cirrhosis (Child-Pugh class C) were assigned to groups given either pentoxifylline (400 mg, orally, 3 times daily; n = 164) or placebo (n = 171) for 6 months. The primary end point was mortality at 2 months. Secondary end points were mortality at 6 months and development of liver-related complications. By 2 months, 28 patients in the pentoxifylline group (16.5%) and 31 in the placebo group (18.2%) had died (P = .84). At 6 months, 50 patients in the pentoxifylline group (30.0%) and 54 in the placebo group (31.5%) had died (P = .75). The proportions of patients without complications (eg, bacterial infection, renal insufficiency, hepatic encephalopathy, or gastrointestinal hemorrhage) were higher in the pentoxifylline group than in the placebo group at 2 months (78.6% vs 63.4%; P = .006) and 6 months (66.8% vs 49.7%; P = .002). The probability of survival without complications was higher in the pentoxifylline group than in the placebo group at 2 and 6 months (P = .04). In multivariate analysis, the factors associated with death were age, the Model for End-Stage Liver Disease score, and presence of early-stage carcinoma. Treatment with pentoxifylline was the only factor associated with liver-related complications. Although pentoxifylline does not decrease short-term mortality in patients with advanced cirrhosis, it does reduce the risk of complications.


The enigma of herpes stromal disease.
Herpes stromal disease is due to direct damage as a result of viral replication, virally induced immune mechanisms, or a combination of the two. Viral replication may have a major initiating role in the production of herpes simplex and herpes zoster induced stromal disease, and steroids may initially be harmful in their treatment. On topical antiviral drugs alone, in patients who never previously had had topical steroids, 14 of 15 cases of herpes simplex induced disciform keratitis responded favourably in an average of 44 days of treatment. This compared with one out of 14 responding if steroids had previously been used, 13 of 14 requiring topical steroids and an average 112 days' treatment. In herpes zoster stromal disease cases 78% had epithelial involvement, 54 of 57 responded to topical antivirals alone without the use of steroids, 2% recurred, and treatment averaged a total of 62 days. If steroids were used alone or in combination with antivirals, there was a 50% recurrence rate and 200 day total treatment duration.


High-Resolution Bisulfite-Sequencing of Peripheral Blood DNA Methylation in Early-Onset and Familial Risk Breast Cancer Patients.
Understanding and explaining hereditary predisposition to cancer has focused on the genetic etiology of the disease. However, mutations in known genes associated with breast cancer, such as BRCA1 and BRCA2, account for less than 25% of familial cases of breast cancer. Recently, specific epigenetic modifications at BRCA1 have been shown to promote hereditary breast cancer, but the broader potential for epigenetic contribution to hereditary breast cancer is not yet well understood. We examined DNA methylation through deep bisulfite sequencing of CpG islands and known promoter or regulatory regions in peripheral blood DNA from 99 patients with familial or early-onset breast or ovarian cancer, 6 unaffected BRCA mutation carriers, and 49 unaffected controls. In 9% of patients, we observed altered methylation in the promoter regions of genes known to be involved in cancer, including hypermethylation at the tumor suppressor PTEN and hypomethylation at the proto-oncogene TEX14. These alterations occur in the form of allelic methylation that span up to hundreds of base pairs in length. Our observations suggest a broader role for DNA methylation in early-onset, familial risk breast cancer. Further studies are warranted to clarify these mechanisms and the benefits of DNA methylation screening for early risk prediction of familial cancers.


Experimental results on discrete-time nonlinear adaptive tracking control of a flexible-link manipulator.
The aim of this paper is to develop and implement a nonlinear adaptive control scheme for a single-link flexible manipulator. The controller is designed based on a discrete-time nonlinear model of the arm. The model is derived by using the forward difference method (Euler approximation). The output redefinition concept is then used so that the associated zero dynamics corresponding to the new output is guaranteed to be exponentially stable. An indirect adaptive linearizing controller is developed for the resulting minimum phase system where the "payload mass" is assumed to be unknown but its upper bound is assumed to be known a priori. The performance of the adaptively controlled closed-loop system is investigated by both numerical simulations and experimental results. The proposed controller is also compared experimentally with those of nonadaptive feedback linearization and conventional proportional derivative (PD) control strategies.


Genotoxic Damage During Brain Development Presages Prototypical Neurodegenerative Disease.
Western Pacific Amyotrophic Lateral Sclerosis and Parkinsonism-Dementia Complex (ALS/PDC) is a disappearing prototypical neurodegenerative disorder (tau-dominated polyproteinopathy) linked with prior exposure to phytogenotoxins in cycad seed used for medicine and/or food. The principal cycad genotoxin, methylazoxymethanol (MAM), forms reactive carbon-centered ions that alkylate nucleic acids in fetal rodent brain and, depending on the timing of systemic administration, induces persistent developmental abnormalities of the cortex, hippocampus, cerebellum, and retina. Whereas administration of MAM prenatally or postnatally can produce animal models of epilepsy, schizophrenia or ataxia, administration to adult animals produces little effect on brain structure or function. The neurotoxic effects of MAM administered to rats during cortical brain development (specifically, gestation day 17) are used to model the histological, neurophysiological and behavioral deficits of human schizophrenia, a condition that may precede or follow clinical onset of motor neuron disease in subjects with sporadic ALS and ALS/PDC. While studies of migrants to and from communities impacted by ALS/PDC indicate the degenerative brain disorder may be acquired in juvenile and adult life, a proportion of indigenous cases shows neurodevelopmental aberrations in the cerebellum and retina consistent with MAM exposure in utero. MAM induces specific patterns of DNA damage and repair that associate with increased tau expression in primary rat neuronal cultures and with brain transcriptional changes that parallel those associated with human ALS and Alzheimer's disease. We examine MAM in relation to neurodevelopment, epigenetic modification, DNA damage/replicative stress, genomic instability, somatic mutation, cell-cycle reentry and cellular senescence. Since the majority of neurodegenerative disease lacks a solely inherited genetic basis, research is needed to explore the hypothesis that early-life exposure to genotoxic agents may trigger or promote molecular events that culminate in neurodegeneration.


A randomized, double-blind, placebo-controlled, parallel-group study of methylphenidate transdermal system in pediatric patients with attention-deficit/hyperactivity disorder.
To evaluate the efficacy and safety of methylphenidate transdermal system compared with placebo, using osmotic-release oral system (OROS) methylphenidate as a reference therapy. We conducted a 7-week, randomized, double-blind, double-dummy, placebo-controlled trial in children diagnosed with attention-deficit/hyperactivity disorder by DSM-IV-TR criteria, within a community setting. The study was conducted from August 2004 to February 2005. Participants were randomly assigned to 1 of 3 treatments: methylphenidate transdermal system patch plus placebo capsule (N = 100), OROS methylphenidate capsule plus placebo patch (N = 94), or placebo capsule plus placebo patch (N = 88). Over 5 weeks, once-daily doses were optimized using 10-, 15-, 20-, and 30-mg methylphenidate transdermal system patches (9-hour wear time) or 18-, 27-, 36-, and 54-mg OROS methylphenidate capsules. Thereafter, optimal treatment doses were maintained for 2 weeks with blinded ratings of attention, behavior, and academic performance occurring at the end of each week. The primary efficacy measure was the clinician-rated ADHD Rating Scale-Version IV (ADHD-RS-IV). Additional measures included teacher, parent, and other clinician rating scales. Safety and tolerability were assessed throughout the study. The mean change from baseline in ADHD-RS-IV scores was greater for participants receiving methylphenidate transdermal system and OROS methylphenidate treatments compared with placebo (p < .0001). Similar results were observed for parent and teacher rating scales. More participants receiving active treatments compared with placebo were rated as improved by clinicians and parents (p < .0001). Adverse events were generally mild or moderate in intensity, and the most common included decreased appetite, nausea, vomiting, and insomnia. The results of this study suggest that the methylphenidate transdermal system is an efficacious treatment option for children with attention-deficit/hyperactivity disorder. clinicaltrials.gov Identifier: NCT00444574.


Platelet-Rich Plasma for Frozen Shoulder: A Case Report.
Frozen shoulder is a glenohumeral joint disorder that movement because of adhesion and the existence of fibrosis in the shoulder capsule. Platelet-rich plasma can produce collagen and growth factors, which increases stem cells and consequently enhances the healing. To date, there is no evidence regarding the effectiveness of platelet-rich plasma in frozen shoulder. A 45-year-old man with shoulder adhesive capsulitis volunteered for this treatment. He underwent two consecutive platelet-rich plasma injections at the seventh and eighth month after initiation of symptoms. We measured pain, function, ROM by the visual analogue scale (VAS), scores from the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire and goniometer; respectively. After first injection, the patient reported 60% improvement regarding diurnal shoulder pain, and no night pain. Also, two-fold improvement for ROM and more than 70% improvement for function were reported. This study suggests the use of platelet-rich plasma in frozen shoulder to be tested in randomized trials.


Increased nuchal translucency in fetuses with a normal karyotype-diagnosis and management: An observational study.
The use of nuchal translucency (NT) in 1992 by Nicolaides et al was a major breakthrough in screening for chromosomal aneuploidies at the end of the first trimester. However, pathological conditions other than chromosomal aneuploidies are also associated with increased NT, which can also be detected in normal fetuses. This study sought to evaluate the causes of this ultrasound sign in a group of patients from Iasi, Romania.During the decade-long study period, there were 71 certified cases involving increased NT; the patients in these cases underwent diagnostic amniocentesis and karyotyping.In most of the examined cases (55 cases, 78%), there was no aneuploidy. The remaining cases involved trisomy 21 (T21) (18%), trisomy 18 (T18) (2%), or triploidy (2%). In most cases, the indication for amniocentesis was increased NT alone (81%), whereas the remaining cases also involved advanced maternal age (5.5%), abnormal serologic markers (10%), or other ultrasound signs (3.5%) (2 cases-cardiac anomalies and fetal hydrops). A favorable pregnancy outcome at term was achieved in 40 cases (56% from total, 72% from euploid pregnancies); kidney anomalies or nonlethal cardiac conditions were observed in 12 cases (17%), 6 of which involved complications associated with premature onset of labor, and miscarriages occurred in 6 cases. Three cases were lost at follow-up.Although it is common practice to assume that increased NT is an indication for amniocentesis, both literature results and our study findings indicate that the majority of cases with increased NT involve no aneuploidy and result in a favorable outcome if no other anomaly is present. Better evidence-based management of such cases could be promoted by conducting larger, multicenter studies, and following cases for longer periods.


[Clinical efficacy of tianeptine in patients with ischemic heart disease and comorbid depression].
Ischemic heart disease patients (20 men, 20 women, age 36-72 years) with class II-III effort angina and depression (Beck Depression Inventory -- BDI -- score > or = 19) were randomized to standard therapy were treatment of stable ischemic heart disease (control group) or standard therapy plus tianeptine 37.5 mg/day. After 6 weeks 52% decrease of BDI score occurred in tianeptine treated patients (from 24.9+/-1.2 to 11.9+/-1.5, p<0.001). This was associated with decrease of number and severity of cardialgias, better blood pressure control in patients with hypertension, lengthening of exercise time during exercise test (by 3.3+/-0.9 min, p<0.05), and increase of overall index of quality of life (by 2.6+/-0.9 points, p<0.01). No dynamics of these parameters occurred in control group.


Racial and Ethnic Disparities in Perceived Barriers to Health Care Among U.S. Adults With Intellectual and Developmental Disabilities.
Barriers to health care access can greatly affect one's health status. Research shows that U.S. adults with intellectual and developmental disabilities (IDD) have poor health and face barriers such as long waits for appointments. However, whether barriers differ by race and ethnicity has not been examined. We conducted a secondary data analysis using the 2002-2011 Medical Expenditure Panel Survey dataset, and compared perceived barriers of community-living U.S. adults with IDD in three racial and ethnic groups (White, Black, and Latinx). Specifically, we examined the top reasons for not having usual source of care, delaying or foregoing medical care. For Black and Latinx adults with IDD, the most-mentioned reasons for not having usual source of care, delaying or foregoing medical care were "don't like/don't trust doctors," "don't use doctors," and "don't know where to get care." In comparison, the White adults with IDD group's biggest perceived barriers were location and insurance related. All groups cited that being unable to afford care was a top reason for delaying or foregoing care. Policies/interventions to improve health care access in racial/ethnic minorities with IDD must first address the topic of developing trust between patients and the health professions. Insurance and the rising costs of care are also key areas that need attention.


CITRATE CLEAVAGE ENZYME IN LIVERS OF OBESE AND NONOBESE MICE.
The specific activity of the citrate cleavage enzyme is 3.3 times greater in livers of obese mice than in livers of their nonobese litter mates. The difference persists during starvation. The specific activity of the acetate activating enzyme is approximately the same in the livers of both types of animals. It is proposed that a high activity of citrate cleavage enzyme is one of the factors responsible for obesity.


BioMaPS: a roadmap for success.
The manuscript outlines the impact that our National Science Foundation Interdisciplinary Training for Undergraduates in Biological and Mathematical Sciences program, BioMaPS, has had on the students and faculty at Murray State University. This interdisciplinary program teams mathematics and biology undergraduate students with mathematics and biology faculty and has produced research insights and curriculum developments at the intersection of these two disciplines. The goals, structure, achievements, and curriculum initiatives are described in relation to the effects they have had to enhance the study of biomathematics.


Attenuation of calcitonin gene expression in pregnant rat uterus leads to a block in embryonic implantation.
The peptide hormone calcitonin plays a key role in calcium homeostasis in many tissues, such as bone and kidney. Our previous studies revealed that the expression of calcitonin is dramatically induced in the glandular epithelium of rat uterus between days 3-5 of pregnancy before the onset of blastocyst implantation on day 5. Calcitonin expression is switched off once implantation has progressed to day 6. The coincidence in timing suggested that calcitonin may function as a regulatory signal in the uterus during the early events leading to implantation. Here we report that the implantation stage-specific expression of calcitonin can be specifically attenuated by administering antisense oligodeoxynucleotides (ODNs) directed against exon IV of calcitonin messenger RNA into the uterine horns on day 2 of gestation. The loss of calcitonin messenger RNA and protein expression upon antisense ODN treatment is accompanied by a severe impairment in implantation of embryos. Based on the observations that 1) treatment with two different antisense ODNs possessing different base compositions produced similar phenotypes; and 2) treatment with the complementary sense ODNs did not affect either calcitonin expression or implantation, we conclude that the effects of antisense ODNs on calcitonin expression and implantation are specific and functionally linked. Our study strengthens the hypothesis that a transient expression of calcitonin in the preimplantation phase uterus is critical for blastocyst implantation.


Using a type 1 collagen-based system to understand cell-scaffold interactions and to deliver chimeric collagen-binding growth factors for vascular tissue engineering.
Vascular tissue engineering should provide more biocompatible and functional conduits than synthetic vascular grafts. Understanding cell-scaffold interactions and developing an efficient delivery system for growth factors and other biomolecules to control the signaling between the cells and the scaffold are fundamental issues in a wide range of tissue engineering research fields. Type 1 collagen is a natural scaffold extensively used in vascular tissue engineering and is a widely used vehicle in biomolecule delivery. In this article, we will discuss type 1 collagen as a vascular tissue engineering scaffold, describe strategies for elucidating the interaction between cells and type 1 collagen scaffolds using various imaging techniques, and summarize our work on the development of a chimeric collagen-binding growth factor-based local delivery system.


Temporal inhibition in character identification.
Models of information processing tasks such as character identification often do not consider the nature of the initial sensory representation from which task-relevant information is extracted. An important component of this representation is temporal inhibition, in which the response to a stimulus may inhibit, or in some cases facilitate, processing of subsequent stimuli. Three experiments demonstrate the existence of temporal inhibitory processes in information processing tasks such as character identification and digit recall. An existing information processing model is extended to account for these effects, based in part on models from the detection literature. These experiments also discriminate between candidate neural mechanisms of the temporal inhibition. Implications for the transient deficit theory of dyslexia are discussed.


Psychiatrists' ascertained treatment needs for mental disorders in a population-based sample.
Population-based studies of prevalence have been used to approximate the amount of need for treatment of mental disorders. This study aimed to estimate need for treatment of alcohol dependence, major depression, panic disorder, agoraphobia, and social phobia. Psychiatrists used the Schedule for Clinical Assessment in Neuropsychiatry (SCAN) to assess individuals in a probability sample of the Baltimore Epidemiologic Catchment Area follow-up study for need for and use of mental health services. Population-based estimates were calculated with probability weights. The SCAN sample included 816 participants who were interviewed between 1993 and 1999. Prevalence of need for mental health services in the general population was 28.7%+/-2.7%. Among the disorders studied, the greatest need was for treatment of alcohol dependence; 90% of the SCAN sample who met criteria for this diagnosis, or 13.6%+/-1.9% of the population, needed treatment. In the general population, 10.5%+/-2.1% needed treatment for major depression; 5.2%+/-.9%, for social phobia; 5.2%+/-1.6%, for panic disorder; and 3.2%+/-.9%, for agoraphobia. The highest needs for specific treatment modalities were self-help groups for alcohol dependence, talk therapy for depression, behavior modification for social phobia and agoraphobia, and antidepressant medications for panic disorder. Overall, less than 33% of needs were met. Social phobia and alcohol dependence were the two disorders with the highest proportion of unmet need. Prevalence of mental disorders is only an approximation of the need for treatment. There is a substantial need for mental health services in the general population. Most individuals who would benefit from treatment are not receiving any type of mental health services.


Structural organization of the 3' half of the rat thyroglobulin gene.
We report the structural organization of an 80 Kb segment of rat DNA, which encodes for about 40% of Thyroglobulin mRNA at the 3' end. The codogenic information included in this segment is splitted in 17 exons of homogeneous size (about 200 bp). The seven exons at the extreme 3' end have been precisely defined by DNA sequence analysis. No clear sequence homology is found among the exons, even though their coding capacity is quite similar, from 55 to 63 aminoacids residues. We located 2 hormonogenic (T4 forming) sites on the extreme 3' end of the gene in different exons. The DNA sequence coding for these functional sites shows a 70% homology in a 50 nucleotides segment. In addition we found a remnant of this sequence in other exons of the gene. Two large introns have been found on the 3' end of the gene: one is 17 Kb and the other one is more than 30 Kb long. On the basis of these findings and of preliminary studies on the remaining 5' end of the gene, we can predict that the minimum length of the rat TGB gene will be 150 Kb, which makes this gene the largest so far identified eukaryotic gene. We propose in addition that the 3' end exons arose by duplication of a common ancestor.


Sequence analysis and expression of the Salmonella typhimurium asr operon encoding production of hydrogen sulfide from sulfite.
A chromosomal locus of Salmonella typhimurium which complements S. typhimurium asr (anaerobic sulfite reduction) mutants and confers on Escherichia coli the ability to produce hydrogen sulfide from sulfite was recently cloned (C. J. Huang and E. L. Barrett, J. Bacteriol. 172:4100-4102, 1990). The DNA sequence and the transcription start site have been determined. Analysis of the sequence and gene products revealed a functional operon containing three genes which have been designated asrA, asrB, and asrC, encoding peptides of 40, 31, and 37 kDa, respectively. The predicted amino acid sequences of both asrA and asrC contained arrangements of cysteines characteristic of [4Fe-4S] ferredoxins. The sequence of asrB contained a typical nucleotide-binding region. The sequence of asrC contained, in addition to the ferredoxinlike cysteine clusters, two other cysteine clusters closely resembling the proposed siroheme-binding site in biosynthetic sulfite reductase. Expression of lacZ fused to the asr promoter was repressed by oxygen and induced by sulfite. Analysis of promoter deletions revealed a region specific for sulfite regulation and a second region required for anaerobic expression. Computer-assisted DNA sequence analysis revealed a site just upstream of the first open reading frame which had significant homology to the FNR protein-binding site of E. coli NADH-linked nitrite reductase. However, asr expression by the fusion plasmid was not affected by site-specific mutations within the apparent FNR-binding site.


Sites on calmodulin that interact with the C-terminal tail of Cav1.2 channel.
Two fragments of the C-terminal tail of the alpha(1) subunit (CT1, amino acids 1538-1692 and CT2, amino acids 1596-1692) of human cardiac L-type calcium channel (Ca(V)1.2) have been expressed, refolded, and purified. A single Ca(2+)-calmodulin binds to each fragment, and this interaction with Ca(2+)-calmodulin is required for proper folding of the fragment. Ca(2+)-calmodulin, bound to these fragments, is in a more extended conformation than calmodulin bound to a synthetic peptide representing the IQ motif, suggesting that either the conformation of the IQ sequence is different in the context of the longer fragment, or other sequences within CT2 contribute to the binding of calmodulin. NMR amide chemical shift perturbation mapping shows the backbone conformation of calmodulin is nearly identical when bound to CT1 and CT2, suggesting that amino acids 1538-1595 do not contribute to or alter calmodulin binding to amino acids 1596-1692 of Ca(V)1.2. The interaction with CT2 produces the greatest changes in the backbone amides of hydrophobic residues in the N-lobe and hydrophilic residues in the C-lobe of calmodulin and has a greater effect on residues located in Ca(2+) binding loops I and II in the N-lobe relative to loops III and IV in the C-lobe. In conclusion, Ca(2+)-calmodulin assumes a novel conformation when part of a complex with the C-terminal tail of the Ca(V)1.2 alpha(1) subunit that is not duplicated by synthetic peptides corresponding to the putative binding motifs.


Sensory neuron fates are distinguished by a transcriptional switch that regulates dendrite branch stabilization.
Sensory neurons adopt distinct morphologies and functional modalities to mediate responses to specific stimuli. Transcription factors and their downstream effectors orchestrate this outcome but are incompletely defined. Here, we show that different classes of mechanosensory neurons in C. elegans are distinguished by the combined action of the transcription factors MEC-3, AHR-1, and ZAG-1. Low levels of MEC-3 specify the elaborate branching pattern of PVD nociceptors, whereas high MEC-3 is correlated with the simple morphology of AVM and PVM touch neurons. AHR-1 specifies AVM touch neuron fate by elevating MEC-3 while simultaneously blocking expression of nociceptive genes such as the MEC-3 target, the claudin-like membrane protein HPO-30, that promotes the complex dendritic branching pattern of PVD. ZAG-1 exercises a parallel role to prevent PVM from adopting the PVD fate. The conserved dendritic branching function of the Drosophila AHR-1 homolog, Spineless, argues for similar pathways in mammals.


Detecting electroporation by assessing the time constants in the exponential response of human skin to voltage controlled impulse electrical stimulation.
We propose a new method for extracting the electrical properties of human skin based on the time constant analysis of its exponential response to impulse stimulation. As a result of this analysis an adjacent finding has arisen. We have found that stratum corneum electroporation can be detected using this analysis method. We have observed that a one time-constant model is appropriate for describing the electrical properties of human skin at low amplitude applied voltages (<30V), and a two time-constant model best describes skin electrical properties at higher amplitude applied voltages (>30V). Higher voltage amplitudes (>30V) have been proven to create pores in the skin's stratum corneum which offer a new, lower resistance, pathway for the passage of current through the skin. Our data shows that when pores are formed in the stratum corneum they can be detected, in-vivo, due to the fact that a second time constant describes current flow through them.


Interaction between bacterial cell membranes and nano-TiO2 revealed by two-dimensional FTIR correlation spectroscopy using bacterial ghost as a model cell envelope.
The interaction between microorganisms and nanoparticles is a crucial step towards understanding the subsequent biological effect. In this study, the interaction between TiO2 nanoparticles and bacterial cell membrane was investigated by Two-dimensional Correlation Fourier Transformation Infrared spectroscopy (2D-FTIR-COS) using bacterial ghosts (BGs), which are non-living bacterial cell envelopes devoid of cytoplasm. The synchronous map of 2D-FTIR-COS results indicated that the functionalities in proteins of BGs preferentially interacted with TiO2 nanoparticles; whereas the interaction of TiO2 nanoparticles with characteristic functionality in polysaccharides (COH) and phospholipids (PO) were very weak or insensitive. This conclusion was further corroborated by settling of TiO2 nanoparticles in the presence of pure protein, polysaccharide and phospholipid represented by bovine serum albumin (BSA), alginate and phosphatidylethanolamine (PE). Additionally, the asynchronous map of 2D-FTIR-COS indicated a sequential order of functionalities bonded to TiO2 nanoparticles with the order of: COO- > aromatic CC stretching > NH, amide II > CO, ketone. These findings contribute to deeper understanding of the interaction between TiO2 nanoparticles and bacterial cell membrane in aquatic systems.


[Comparative study of the effectiveness of ethacizine, etmozin and Ritmilen in patients with ventricular rhythmic disorders].
Antiarrhythmic effect of ethacizine, a new Soviet compound, was compared to those of etmozin and Ritmilen. Ethacizine was given to 30 patients with chronic ventricular rhythm disorders, including those with frequent extrasystoles, bigeminy, paired extrasystoles and runs of ventricular tachycardia. Coronary patients prevailed in the sample. Holter's ECG monitoring was used for the assessment of treatment efficiency. Ethacizine showed the best anti-arrhythmic effect (81%), suppressing frequent extrasystoles as well as high-grade extrasystoles in the highest proportion of cases. The lowest efficiency (38%) in combination with the highest incidence of side effects were associated with Ritmilen, while etmozin kept somewhat in between (58% response rate).


Rehabilitation following zone II flexor tendon repairs.
Ongoing clinical and basic research has improved understanding of flexor tendon mechanics and physiology for surgical repair and rehabilitation after a zone II flexor tendon repair. Yet, the ideal surgical repair technique that includes sufficient strength to allow safe immediate active motion of the finger, without excessive repair stiffness, bulk or rough surfaces resulting in excessive resistance to flexion, does not exist. After optimizing the repair, the surgeon and therapist team must select a rehabilitation plan that protects the repair but helps to maintain tendon gliding. There are 3 types of rehabilitation programs for flexor tendon repairs: delayed mobilization, early passive mobilization, or an early active mobilization. No guideline for rehabilitation should be followed exactly. Many factors influence therapy decisions, including repair technique, associated tendon healing, passive versus active range of motion, edema, and tendon adhesions. These factors can assist in guiding rehabilitation progression and promote functional range of motion, safely mobilize the repaired tendon(s) and prevent gapping, rupture, and adhesions.


Emergency Intracranial Stenting in Acute Stroke: Predictors for Poor Outcome and for Complications.
Background Stent-retriever thrombectomy is the first-line therapy in acute stroke with intracranial large vessel occlusion. In case of failure of stent-retriever thrombectomy, rescue stent angioplasty might be the only treatment option to achieve permanent recanalization. This study aims at identifying predictors for poor outcome and complications in a large, multicenter cohort receiving rescue stent angioplasty. Methods and Results We performed a retrospective analysis of patients with large vessel occlusion who were treated with rescue stent angioplasty after stent-retriever thrombectomy between 2012 and 2018 in 7 neurovascular centers. We defined 2 binary outcomes: (1) functional clinical outcome (good modified Rankin Scale, 0-2; and poor modified Rankin Scale, 4-6) and (2) early symptomatic intracerebral hemorrhage. Impacts of clinical, radiological, and interventional parameters on outcomewere assessed in uni- and multivariable logistic regression models. Two hundred ten patients were included with target vessels located within the anterior circulation (136 of 210; 64.8%) and posterior circulation (74 of 210; 35.2%). Symptomatic intracerebral hemorrhage occured in 22 patients, 86.4% (19 of 22) after anterior and 13.6% (3 of 22) after posterior circulation large vessel occlusion. Good functional outcome was observed in 44.8% (73 of 163). A higher National Institutes of Health Stroke Scale on admission (adjusted odds ratio, 1.10; P=0.002), a higher premorbid modified Rankin Scale (adjusted odds ratio, 2.02; P=0.049), and a modified Thrombolysis in Cerebral Infarction score of 0 to 2a after stenting (adjusted odds ratio, 23.24; P<0.001) were independent predictors of poor functional outcome. Conclusions Use of rescue stent angioplasty can be considered for acute intracranial large vessel occlusion in cases after unsuccessful stent-retriever thrombectomy. Likelihood of symptomatic intracerebral hemorrhage is higher in anterior circulation stroke.