The helical content of the YscP molecular ruler determines the length of the Yersinia injectisome.
The length of the Yersinia injectisome needle is determined by the protein YscP, which could act as a molecular ruler. The analysis of the correlation between the size of YscP and the needle length in seven wild-type strains of Yersinia enterocolitica reinforced this hypothesis but hinted that the secondary structure of YscP might influence needle length. Hence, 11 variants of YscP(515) were generated by multiple Pro or Gly substitutions. The needle length changed in inverse function of the helical content, indicating that not only the number of residues but also their structure controls length. Taking the secondary motifs into account, Pro/Gly-variants were subjected to in silico modelling to simulate the extension of YscP upon needle growth. The calculated lengths when the helical content is preserved correlated strikingly with the measured needle length, with a constant difference of approximately 29 nm, which corresponds approximately to the size of the basal body. These data support the ruler model and show that the functional ruler has a helical structure.


A uniform labeling system for bone marrow and peripheral blood stem cells.
The separation of bone marrow (BM) constituents, BM cell purging, and cryopreservation of BM and peripheral blood stem cells (PBSC) are frequently performed in the clinical laboratories. In 1991, recognizing the increasing involvement of transfusion services in BM and PBSC processing, the American Association of Blood Banks (AABB) established standards that, in general, held BM and PBSC processing to the same standards as applied to blood components. In 1993, the AABB defined the information that was required on the container label, but did not establish a uniform system of labeling for BM/PBSC preparations. We present a system of labeling for definitive identification of allogeneic BM and autologous BM/PBSC collections that identifies all components derived from the original product from the time of collection to final infusion or disposal. This system avoids the risk of component misidentification, particularly when multiple collections are processed simultaneously. Our transfusion service computer system was adapted with BM/PBSC component names and abbreviated codes to provide a singular validated mechanism for labeling, tracking, and final disposition of these products. We propose the adaptation of a "universal system" for BM/PBSC labeling for AABB accredited transfusion services utilizing the existing guidelines for labeling of blood and blood components.


Levels of serum protease inhibitors during fetal and postnatal development of the pig.
Using electroimmunoassay the levels of the individual protease inhibitors alpha-2-macroglobulin f, alpha-2-macroglobulin s, inter-alpha-trypsin inhibitor, alpha-1-protease inhibitor and alpha-2-antitrypsin were studied in the serum and the amniotic fluid during the fetal and postnatal development of the pig. High concentrations, far above the adult ones, of alpha-2-antitrypsin and especially alpha-1-protease inhibitor, were observed in serum during the fetal period, contrary to the low levels of the alpha-macroglobulins. Thus, the development of these inhibitors in pigs differs markedly from the development in the human fetus. After birth, when the piglet had begun to suckle, the levels of all the inhibitors, except that of alpha-2-macroglobulin s, decreased. This can partly be explained by the dilution of the serum due to the increase in plasma volume and losses via the urine. The sharp increase of alpha-2-macroglobulin s during the first days after birth probably represented an extensive synthesis. All the inhibitors had attained their adult levels at about 35 days after birth. All the protease inhibitors studied, except for alpha-2-macroglobulin s, were found in the amniotic fluid, and reached the maximal levels at about 55-66 days of gestation. These inhibitors appeared to have fetal origin. The physiological significance of the protease inhibitors during fetal and neonatal development of the pig is discussed. it is suggested that the function of these inhibitors might be to protect the cells and tissues from proteolytic attack by the formation of inactive enzyme/inhibitors complexes.


Cellular and Extracellular Components in Tumor Microenvironment and Their Application in Early Diagnosis of Cancers.
Tumors are surrounded by complex environmental components, including blood and lymph vessels, fibroblasts, endothelial cells, immune cells, cytokines, extracellular vesicles, and extracellular matrix. All the stromal components together with the tumor cells form the tumor microenvironment (TME). In addition, extracellular physical and chemical factors, including extracellular pH, hypoxia, elevated interstitial fluid pressure, and fibrosis, are closely associated with tumor progression, metastasis, immunosuppression, and drug resistance. Cellular and extracellular components in TME contribute to nearly all procedures of carcinogenesis. By summarizing the recent work in this field, we make a comprehensive review on the role of cellular and extracellular components in the process of carcinogenesis and their potential application in early diagnosis of cancer. We hope that a systematic review of the diverse aspects of TME will help both research scientists and clinicians in this field.


Development of Antimicrobial Packaging Film Made from Poly(Lactic Acid) Incorporating Titanium Dioxide and Silver Nanoparticles.
Polylactide (PLA)/nano-TiO₂ and PLA/nano-TiO₂/nano-Ag blends films were prepared by a solvent volatilization method. Compared to pure PLA film, the nano-blend films have low water vapor permeability (WVP) and a poor transparency. With the increase of the NPs in the PLA, the tensile strength (TS) and elastic modulus (EM) decreased, while the elongation at break (ε) increased. SEM analysis indicated a rougher cross-section of the nano-blend films. According to the FTIR analysis, no new chemical bonds were formed in the nano-blend films. By using DSC to examine the crystallization and melting behavior, the result shows that the NPs have no effect on the glass transition (Tg) and melting temperature (Tm), but they caused an increase on the cold crystallization (Tc) and crystallinity (Xc). TGA results show that the addition of nanoparticles significantly improved the thermal stability. The PLA nano-blend films show a good antimicrobial activity against. E. coli and Listeria monocytogenes. Most important, we carried out migration tests, and verified that the release of NPs from the nano-blend films was within the standard limits.


Nursing students' willingness to use complementary and alternative therapies for cancer patients: Istanbul survey.
It is important for student nurses to be knowledgeable of the complementary and alternative therapies and to provide accurate information to both cancer patients and other health care professionals. This study examined the nursing students' willingness to use these therapies, availability of sources of information, use of the therapies for self care, opinions about the integration of these therapies into nursing curriculum, and analyzed the differences among the responses. A self-administered questionnaire was offered to 640 nursing students in Istanbul, descriptive statistics were used, and comparisons among responses were made with chi-square test. Willingness to use for cancer patients was highest for nutritional therapy (76.1%), breathing therapies (74.5%), and massage and manipulation-Tui Na, in which pressure and touch are applied to the body (71.9%). Use of information sources was highest for nutritional therapy (75.6%), breathing therapies (71.9%), and massage and manipulation-Tui Na (62.3%). Over half of the nursing students used music therapy (54.2%), and massage and manipulation-Tui Na (53.6%) for self-care. Breathing therapies (87.2%) were the most desired therapy chosen to be included in nursing curriculum. The statistically significant differences were found among the responses related to use five therapies for care and related to desired three therapies to be included in nursing curriculum. Although students had not previously been exposed to these therapies use with oncology patients, many of students expressed a desire to integrate therapies learning into nursing curriculum. The more student nurses document high risk patients, the more effective strategies will be developed by other health care professionals.


Understanding the experience of women admitted to a psychiatric hospital in Sydney with psychosis or mania following childbirth after World War II (1945-1955).
In the present study, we investigated a unique set of historical health-care records of women admitted to a psychiatric hospital in Sydney, Australia with a diagnosis of psychosis or mania after childbirth in the post-World War II (WWII) period, from 1945 to 1955. This research is part of a larger project examining how the descriptions of these women documented in the health-care records from 1885 to 1975 affected their treatment and the outcome of their admission. In the present paper, we report on the findings from an intensive examination of the post WWII documents. Eighteen health-care records from a psychiatric facility (Gladesville Hospital) were identified from admission registers housed in the State Records Office of New South Wales in 2014. Although seven records had been destroyed, 11 were transcribed verbatim. The records contain demographic information; descriptions of the women's signs and symptoms on admission; and information about the women before, during, and after their admission found in letters from relatives or medical staff. A content analysis of admission information showed how the women were described by health-care professionals, but a textual analysis of the records revealed that there were other factors that could have contributed to the women's condition, which might not have been taken into consideration when treatment and care were devised. The present study demonstrates the value of investigating historical health-care records to understand how prevailing attitudes and practices might affect diagnosis and treatment.


Pituitary hyperplasia.
Pituitary hyperplasia is rare, difficult to diagnose and sometimes controversial. The hyperplasia could be physiologic which is usually reversible, or pathologic which varies in presentation from incidental to tumor like lesion with and without hormonal disturbance. Any pituitary cell is capable of undergoing hyperplasia in the presence of the right stimuli. In this article we summarize the various pathologic and morphologic features of each subtype of pituitary hyperplasia, give an account of the molecular, hormonal and cellular basis of this condition and outline its clinical significance, differential diagnosis and prognosis.


Potentiation of infectivity and pathogenesis of influenza A virus by a house dust mite protease.
Common house dust mites (e.g., Dermatophagoides farinae) excrete a serine-type (Df) protease. Df protease obtained from cultured mites enhanced viral replication in vitro via proteolytic cleavage of viral hemagglutinin (HA) into HA1 and HA2, which confers potent viral infectivity. Its potency is 2- to 5-fold higher than bovine trypsin or human plasmin. Df protease also markedly accelerated virus propagation in vivo: A minute quantity of protease (estimated delivered amount, 0.8-3.2 micrograms) produced approximately 4- to 100-fold increases in infectious virus in the mouse lung. Similar augmentation of viral replication by Df protease was observed in ferret models of nasopharyngeal infections of influenza virus. All extracts from ordinary house dust contained a serine-type protease that cleaved HA into HA1 and HA2. Thus, mite protease in house dust may enhance the pathogenesis of influenza virus.


Soil microbial response to glucose and phosphorus addition under agricultural systems in the Brazilian Cerrado.
Conventional tillage (CT) and no-tillage (NT) management systems alter soil nutrient availability and consequently modify soil microbial response to nutrient additions such as carbon (C) and phosphorus (P). The objective of this study is to evaluate microbial response to the addition of C (glucose) and P (Na2HPO4.7H2O) under CT and NT in the brazilian Cerrado. In response to glucose addition, the NT system yielded higher microbial respiration rates and glucose consumption than the CT system. The best microbial response to C addition was after 0 - 12 h incubation in NT and 0 - 24 h in CT. The addition of P produced higher demand under CT than NT. After incubation, biochemical indicators such as microbial respiration, glucose consumption, dehydrogenase activity and metabolic yield confirmed the higher glucose demands under NT and higher phosphorus demands under CT. These results demonstrate that C and P addition alter significantly the microbial response, suggesting that soil microorganisms present nutrient differential demands between CT and NT management systems.


[Gene Mutation Spectrum Analysis of 170 Patients with Duchenne/Bayesian Muscular Dystrophy in Southwest of China].
To determine gene variations and genotype-phenotype correlations in Duchenne/Bayesian muscular mystrophy (DMD/BMD) patients, and the association between dystrophin gene polymorphisms and clinical phenotype. Multiplex ligation-dependent probe amplification (MLPA) was adopted to detect dystrophin gene variations in 170 patients. Sanger sequencing was performed in 3 cases with decreased peaks in MLPA results. The MLPA detected 72.94% mutations in dystrophin gene, including 62.35% (106/170) deletions, 8.82% (15/170) duplications, and 1.76% (3/170) point mutations. 64 different types of mutations were found. 75.47% of deletions occurred in the range from exon 44 to 55. Most 5' breakpoints of exonic variations were located in 2 hotspots (major hotspot: intron 43-55; minor hotspot: intron 1-20), which is different from findings of other studies. Genotype-phenotype analysis showed that the severity of DMD/BMD was associated with frame shift mutation (r = 0.640, P < 0.001) but not with deletions or duplications. Deletions and duplications of exon compose the main type of dystrophin gene mutations. DMD/BMD is associated with frame shift mutation.


Contribution of anti-inflammatory/immune suppressive processes to the pathology of sepsis.
Sepsis is the leading cause of death in critically ill patients in the United States. It is associated with enormous expenditures within the health care system and despite substantial human, medical and fiscal resources directed at this clinical entity we have only had a modest effect on the septic patient's long-term survival. However, extensive studies over the last few decades have begun to reveal important pathophysiological processes around which a few promising therapeutic strategies with potential benefits may be derived. It is generally believed, that the body reacts to a septic challenge with an intense hyper-inflammatory response, designed to eliminate the underlying pathogen. However, along with and in response to the intense pro-inflammatory reaction, mechanisms fall into place to counter regulate (control) this initial response, typically resulting in a down regulation of the inflammatory response. This frequently results in dysfunction of various immunological conditions and may result in the inability to ward off the infection and consecutively lead to multiple organ dysfunction, multiple organ failure and death. It is the aberrant development of this anti-inflammatory/ immunosuppressive response, in which it is important to expand our understanding of pathological components to develop potential remedy. Upon this background this review aims to provide an overview on the pathophysiological mechanisms which initiate or maintain the down regulation of the immune response to a septic challenge and which might be a starting point for the development of therapeutic strategies.


Rydberg energies using excited state density functional theory.
We utilize excited state density functional theory (eDFT) to study Rydberg states in atoms. We show both analytically and numerically that semilocal functionals can give quite reasonable Rydberg energies from eDFT, even in cases where time dependent density functional theory (TDDFT) fails catastrophically. We trace these findings to the fact that in eDFT the Kohn-Sham potential for each state is computed using the appropriate excited state density. Unlike the ground state potential, which typically falls off exponentially, the sequence of excited state potentials has a component that falls off polynomially with distance, leading to a Rydberg-type series. We also address the rigorous basis of eDFT for these systems. Perdew and Levy have shown using the constrained search formalism that every stationary density corresponds, in principle, to an exact stationary state of the full many-body Hamiltonian. In the present context, this means that the excited state DFT solutions are rigorous as long as they deliver the minimum noninteracting kinetic energy for the given density. We use optimized effective potential techniques to show that, in some cases, the eDFT Rydberg solutions appear to deliver the minimum kinetic energy because the associated density is not pure state v-representable. We thus find that eDFT plays a complementary role to constrained DFT: The former works only if the excited state density is not the ground state of some potential while the latter applies only when the density is a ground state density.


The geographical distribution of Q fever.
The results of a WHO-assisted survey of the distribution of Q fever in 32 countries and an analysis of reports published to date indicate that Q fever exists in 51 countries on five continents. Q-fever infection was most often reported in man and the domestic ruminants, such as cattle, sheep, and goats.The disease was found to exist in most countries where investigations were carried out. Notable exceptions were Ireland, the Netherlands, New Zealand, Poland, and the Scandinavian countries. With the exception of Poland, where the results were inconclusive, all these countries import relatively few domestic ruminants-the most important animal reservoirs of human Q-fever infection. It seems, therefore, that the traffic of infected ruminants may be one of the most important, if not the most important, means for the geographical spread of Q fever. The importance, if any, of ticks associated with such traffic needs to be defined.


Some considerations for the development of diets for mature rodents used in long-term investigations.
Nutritional requirements for mature rodents used in long-term investigations are virtually unknown. The limited knowledge of the dietary needs of mature rodents is due in part to overreliance on weanling animals fed an experimental diet for relatively short periods. Generalizations made from observations of weanling rodents are not appropriate for all ages. Dietary recommendations for rodents have been established, for the most part, by using the nutritional benchmark of maximal growth rate in animals fed ad libitum. Although this method provides valuable insight into the understanding of nutritional deficiency, it is less effective in determining nutrient requirements for mature animals used for the long term. The implication that maximal growth resulting from ad libitum feeding may not indicate the best dietary regimen in the long term is consistent with the observation that energy-restricted rodents live significantly longer and have lower incidence of disease that do their ad libitum-fed counterparts. These and other findings discussed in the review suggest that nutrient requirements established for young rodents may need re-evaluation to determine their applicability to the dietary recommendations for older animals used in long-term investigations.


Simultaneous positioning of cells into two-dimensional arrays using ultrasound.
Contactless simultaneous positioning of micrometer-sized particles in suspension (e.g., copolymer beads, living cells, silicon microparts) can be performed using ultrasound. Current devices are capable of collecting particles into planes or lines by exciting a resonance in the fluid by means of a piezoelectric transducer located beneath the fluidic cavity and are designed such that a one-dimensional pressure field is created. The focus of this work is to collect cells in distinct point locations for potential drug screening array applications. A device to create two-dimensional arrays of cells within a micromachined chamber is described. The chamber is etched into a silicon wafer and sealed with glass; on the underside of the silicon layer a piezoelectric actuator is attached. A signal is applied to each of two orthogonally aligned strips electrodes defined on the surface of the piezoelectric plate. These two strip electrodes create independently addressable approximately one-dimensional pressure fields. It is shown that by applying the same signal to each electrode a diagonally aligned grid of cells can be produced. However, the independence of the two electrodes allows the application of two signals with slightly different frequencies to be applied which creates a grid of circular cell clumps highly suitable for the identified application. (c)


Leukemia derived growth factors produced by human malignant T-lymphoid cell lines.
An autocrine (noninterleukin 2) growth factor, which we term leukemia derived growth factor (LDGF), has previously been found in the culture supernatant of the human malignant T-lymphoid cell line MOLT-4f. We now show that two other human malignant T-lymphoid cell lines, CCRF-CEM and CCRF-HSB-2 also produce such a factor. All three factors, i.e., the LDGF from MOLT-4f, CCRF-CEM, and CCRF-HSB-2 are similar to each other both in biological activity and in physicochemical characteristics. In addition to their autocrine activity, these LDGFs stimulate the growth of other malignant T-lymphoid cell lines, but they do not stimulate B-lymphoblastoid or myeloid cell lines. The results therefore suggest that these LDGFs are T-cell specific.


Y-chromosome STR haplotypes in Danes.
A total of 185 unrelated Danish males were typed for the Y-chromosome STRs DYS19, DYS385a/b, DYS389-I, DYS389-II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438 and DYS439 using the kits PowerPlex Y (Promega), ReliaGene Y-Plex 6 and ReliaGene Y-Plex 5 (Reliagene Technologies). A total of 163 different haplotypes were observed and among these, 144 haplotypes were unique. The gene diversity was 0.9985. In DYS392, a variant allele migrating as a 10.2 allele was observed. Sequencing of the allele showed a deletion upstream the repeated area.


The relationship between online learning self-efficacy, informal digital learning of English, and student engagement in online classes: the mediating role of social presence.
This study investigates the interplay between Online Learning Self-Efficacy (OLSE), Informal Digital Learning of English (IDLE), student engagement in online classes, and the mediating effect of Social Presence (SP) among intermediate Chinese English as a Foreign Language (EFL) students. The research sample consisted of 389 participants enrolled in online English courses within a chain of language schools. Data collection involved the use of validated scales to assess OLSE, IDLE, SP, and online student engagement. Structural Equation Modeling (SEM) was employed for data analysis. The findings revealed direct influences of OLSE and SP on online student engagement. Additionally, IDLE exhibited a positive impact on online student engagement, with a partial mediation effect through SP. This suggests that informal digital English learning significantly contributes to students' engagement in online classes, with this influence being facilitated by the sense of social presence experienced by students during virtual interactions. This research underscores the importance of OLSE, IDLE, and SP in shaping student engagement within online learning environments. The results highlight that fostering informal digital English learning can enhance students' active participation in online courses, with SP serving as a key mediator in this relationship. These insights provide valuable guidance for educators and institutions seeking to improve student engagement in online educational settings.


The Relationship Between Hearing and Mild Behavioral Impairment and the Influence of Sex: A Study of Older Adults Without Dementia from the COMPASS-ND Study.
Hearing loss and mild behavioral impairment (MBI), both non-cognitive markers of dementia, can be early warning signs of incident cognitive decline. We investigated the relationship between these markers and reported the influence of sex, using non-dementia participants (n = 219; 107 females) from the Canadian Comprehensive Assessment of Neurodegeneration and Dementia (COMPASS-ND). Hearing was assessed with the 10-item Hearing Handicap for the Elderly-Screening (HHIE-S) questionnaire, a speech-in-noise test, screening audiometry, and hearing aid use. MBI symptoms were assessed using the Neuropsychiatric Inventory Questionnaire (NPI-Q). Multivariable linear regressions examined the association between hearing and MBI symptom severity and multiple logistic regressions examined the association between hearing and MBI domains. HHIE-S score was significantly associated with greater global MBI symptom burden, and symptoms in the apathy and affective dysregulation domains. Objective measures of audiometric hearing loss and speech-in-noise testing as well as hearing aid use were not associated with global MBI symptom severity or the presence of MBI domain-specific symptoms. Males were older, had more audiometric and speech-in-noise hearing loss, higher rates of hearing-aid use, and showed more MBI symptoms than females, especially apathy. The HHIE-S, a subjective self-report measure that captures emotional and social aspects of hearing disability, was associated with informant-reported global MBI symptom burden, and more specifically the domains of affective dysregulation and apathy. These domains can be potential drivers of depression and social isolation. Hearing and behavior change can be assessed with non-invasive measures, adding value to a comprehensive dementia risk assessment.


Aerolysin of Aeromonas sobria: evidence for formation of ion-permeable channels and comparison with alpha-toxin of Staphylococcus aureus.
Aerolysin from Aeromonas sobria AB3 was isolated and purified. The pure toxin formed sodium dodecyl sulfate-insoluble oligomers in a lipidic environment. The addition of aerolysin to the aqueous phase bathing lipid bilayer membranes resulted in the formation of ion-permeable channels which had a single-channel conductance of about 70 pS in 0.1 M KCl. This defines the toxin as a channel-forming component similar to other toxins but without any indication for an association-dissociation reaction, since the channels had a long lifetime at low voltages. At voltages higher than 50 mV, the aerolysin channel switched into a closed state with a low residual conductance. The single-channel conductance was a linear function of the total aqueous conductance, which suggested that the toxin oligomers formed aqueous channels with an estimated minimal diameter of about 0.7 nm. The aerolysin pores were found to be slightly anion selective. The pore-forming properties of aerolysin were compared with those of alpha-toxin of Staphylococcus aureus. Both aerolysin and alpha-toxin share secondary structure features, must oligomerize to form pores in lipid bilayer membranes, and form channels with similar properties.


Chromatic Information and Feature Detection in Fast Visual Analysis.
The visual system is able to recognize a scene based on a sketch made of very simple features. This ability is likely crucial for survival, when fast image recognition is necessary, and it is believed that a primal sketch is extracted very early in the visual processing. Such highly simplified representations can be sufficient for accurate object discrimination, but an open question is the role played by color in this process. Rich color information is available in natural scenes, yet artist's sketches are usually monochromatic; and, black-and-white movies provide compelling representations of real world scenes. Also, the contrast sensitivity of color is low at fine spatial scales. We approach the question from the perspective of optimal information processing by a system endowed with limited computational resources. We show that when such limitations are taken into account, the intrinsic statistical properties of natural scenes imply that the most effective strategy is to ignore fine-scale color features and devote most of the bandwidth to gray-scale information. We find confirmation of these information-based predictions from psychophysics measurements of fast-viewing discrimination of natural scenes. We conclude that the lack of colored features in our visual representation, and our overall low sensitivity to high-frequency color components, are a consequence of an adaptation process, optimizing the size and power consumption of our brain for the visual world we live in.


Gynecologic Malignancies Post-LeFort Colpocleisis.
Introduction. LeFort colpocleisis (LFC) is a safe and effective obliterative surgical option for older women with advanced pelvic organ prolapse who no longer desire coital activity. A major disadvantage is the limited ability to evaluate for post-LFC gynecologic malignancies. Methods. We present the first case of endometrioid ovarian cancer diagnosed after LFC and review all reported gynecologic malignancies post-LFC in the English medical literature. Results. This is the second reported ovarian cancer post-LFC and the first of the endometrioid subtype. A total of nine other gynecologic malignancies post-LFC have been reported in the English medical literature. Conclusions. Gynecologic malignancies post-LFC are rare. We propose a simple 3-step strategy in evaluating post-LFC malignancies.


Cocaine selling among urban black and white adolescent males.
Data from a longitudinal study of juvenile detainees are used to examine the relationships between cocaine selling, substance use, and other delinquency among the Black and White males in the study. A descriptive comparison of rates of cocaine selling among the youths is followed by a descriptive comparison of prevalence of substance use and other delinquency across four subgroups: White and Black males indicating they sold and did not sell cocaine. These comparisons are followed by analyses of variance examining the relationships between involvement in substance use and other delinquency, and ethnicity, cocaine selling, and the interaction of ethnicity and cocaine selling. Important ethnicity and cocaine-selling effects are found, but not ethnicity by cocaine-selling interactions. The implications of our findings for theory and service provision are drawn.


Defining and Promoting Pediatric Pulmonary Health: Understanding Sleep and Ventilatory Health.
Healthy sleep and optimal ventilatory control begin in early development and are crucial for positive child outcomes. This paper summarizes information presented at the Sleep and Ventilatory Control sessions of the National Heart, Lung, and Blood-sponsored 2021 Defining and Promoting Pediatric Pulmonary Health workshop. These sessions focused on pediatric sleep health, screening for sleep health and sleep disorders in primary care using the electronic health record, infant sleep and ventilatory control, and home sleep testing. Throughout this summary, we discuss key gaps in and barriers to promoting sleep and ventilatory health that were identified during the workshop sessions. We conclude with strategies to address these gaps and barriers and directions for future multidisciplinary research, patient care, and training.


[Characterization of premature ventricular contraction initiating ventricular fibrillation].
The aim of this study is to characterize the electrocardiographic features of premature ventricular contractions (PVC) from different anatomical region that trigger ventricular fibrillation (VF). 36 consecutives patients (20 males, 42+/-14 yrs) undergoing VF ablation from 7 centres were studied (22 with idiopathic VF, 4 associated with a long QT syndrome, 3 with Brugada syndrome, 4 with ischaemic cardiomyopathy and 3 associated with other substrate). Mapping of these PVC showed 2 different origins, which were then confirmed by ablation: right ventricular outflow tract (RVOT) (22%) and peripheral Purkinje network (81%). One patient had PVC from both origins (Brugada). RVOT PVC were frequent but had triggered only 5+/-5 episodes of VF for 26+/-33 months. Purkinje PVC were more likely to be present during electrical storm with 18+/-28 episodes of VF for 33+/-45 months. Right Purkinje PVC have a left bundle branch block with superior axis morphology whereas left Purkinje ones have a right bundle branch block. The axis of activation showed variation from inferior to superior depending on the area of origin from the Purkinje network and the exit site to the myocardium. However Purkinje PVC were characterized by short QRS duration (126+/-18 vs 145+/-13ms for RVOT PVC; p=0.05). In addition the coupling interval was significantly shorter compared to RVOT PVC (292+/-45 vs 358+/-37ms respectively; p=0.005). PVC initiating VF demonstrate specific electrocardiographic features that facilitate determination of their origin. Ablation of these typical PVC is feasible in order to reduce ICD shock.


[Effects of Parkinson's disease on quality-of-life patients' spouses: a qualitative survey].
Spouses play a major role as care givers for their partners with Parkinson's disease. This de facto part of family nursing turns out to be so demanding that they often feel isolated. While spouses may have access to financial and technical aids, but no specific psychological support is available to assist them in coping with the difficulties they have to face. Supporting and educating spouses thus appears today to be a real need. Wishing to create an appropriate support program responding to the needs and expectancies of spouses of Parkinson's disease, we conducted a study designed to measure the effects of Parkinson's disease on spouses' quality-of-life and identify the priority needs in terms of information and support. This study included the spouses of 14 patients who participated in semi-directive individual interviews and a focus group. The data collected shows that spouses experience great disarray when faced with the disease. Their perception of Parkinson's disease has a strong anxiogenic effects. Caring for their spouse on a day to day basis creates a permanent atmosphere of stress with an insecure feeling generating tensions and major frustrations. Most of the spouses do not allow themselves any break and are overwhelmed with ambivalent feelings. They experience a kind of hostility towards their spouse and at the same time feel guilty for their attitude and also for their helplessness. The disease also leads to an impoverishment of the couples' social network, due to reduced autonomy and fear of other people's way of looking at them. Our study confirms the usefulness of organizing an educational support program for these spouses who often feel very lonely and helpless when confronted with their partner's disease.


Modified abaxial approach for resection of the distal sesamoid bone and distal interphalangeal joint while preserving the digital flexor tendons of three Angus bulls with septic arthritis.
CASE DESCRIPTION 3 Angus bulls, aged 2 to 3 years, with severe lameness of 2 to 4 weeks' duration and swelling proximal to the coronary band of the affected limb were evaluated after failing to respond to antimicrobial treatment. CLINICAL FINDINGS Septic arthritis of a distal interphalangeal joint (DIPJ) was diagnosed in all 3 bulls on the basis of results of a physical examination, radiographic and ultrasonographic evaluations of the affected foot, and cytologic evaluation of synovial fluid from the affected DIPJ. TREATMENT AND OUTCOME A novel modified abaxial approach was used to resect the infected distal sesamoid bone (navicular bone) and DIPJ of all 3 bulls. A window was created in the abaxial hoof wall that was lateral to and of sufficient size to extract the navicular bone. Following removal of the navicular bone, the DIPJ was debrided and resected and an orthopedic block was applied to the contralateral claw to minimize weight bearing on the infected digit. Two bulls also had a fiberglass cast applied to the affected limb to help immobilize the DIPJ. All 3 bulls recovered without complications, and 2 bulls were no longer lame, whereas the remaining bull was only mildly lame, at 4 to 5 weeks after surgery. CLINICAL RELEVANCE The modified abaxial approach described for surgical resection of the DIPJ allowed extraction of the infected navicular bone without damage to the digital flexor tendons, something that cannot be achieved with other abaxial approaches. This approach is best used for patients without septic tenosynovitis.


ACTN1 mutations cause congenital macrothrombocytopenia.
Congenital macrothrombocytopenia (CMTP) is a heterogeneous group of rare platelet disorders characterized by a congenital reduction of platelet counts and abnormally large platelets, for which CMTP-causing mutations are only found in approximately half the cases. We herein performed whole-exome sequencing and targeted Sanger sequencing to identify mutations that cause CMTP, in which a dominant mode of transmission had been suspected but for which no known responsible mutations have been documented. In 13 Japanese CMTP-affected pedigrees, we identified six (46%) affected by ACTN1 variants cosegregating with CMTP. In the entire cohort, ACNT1 variants accounted for 5.5% of the dominant forms of CMTP cases and represented the fourth most common cause in Japanese individuals. Individuals with ACTN1 variants presented with moderate macrothrombocytopenia with anisocytosis but were either asymptomatic or had only a modest bleeding tendency. ACTN1 encodes α-actinin-1, a member of the actin-crosslinking protein superfamily that participates in the organization of the cytoskeleton. In vitro transfection experiments in Chinese hamster ovary cells demonstrated that altered α-actinin-1 disrupted the normal actin-based cytoskeletal structure. Moreover, transduction of mouse fetal liver-derived megakaryocytes with disease-associated ACTN1 variants caused a disorganized actin-based cytoskeleton in megakaryocytes, resulting in the production of abnormally large proplatelet tips, which were reduced in number. Our findings provide an insight into the pathogenesis of CMTP.


In Sickness and in Prison: The Case for Removing the Medicare Exclusion for Australian Prisoners.
Many current and former prisoners experience significantly higher rates of physical and mental health problems than others in the community, and are among the most marginalised and disadvantaged people in society. This article argues that granting prison health services an exemption under s 19(2) of the Health Insurance Act 1973 Cth would make the Medicare Benefits Schedule and the Pharmaceutical Benefits Scheme-funded services available to prisoners who meet the eligibility criteria. Australian prisoners would then receive a level of care at least equivalent to that offered by community health services. Reducing health inequities that prisoners experience, particularly Indigenous prisoners, is essential for them continuing to receive health care following release and successfully reintegrating into the community. Further, granting the exemption would assist the Australian Government to meet its international human rights obligations to provide equitable health care for all Australians.


Polyadenylated RNA sequences which are reduced in concentration following auxin treatment of soybean hypocotyls.
Previous work has shown that any effect of exogenous auxin on gene expression in soybean hypocotyl tissue must be restricted to a relatively small fraction of the polyadenylated RNA. However, kinetic hybridization analysis with cDNA probes revealed that a minor abundant class of sequences is markedly reduced in concentrations in the auxin-treated polyadenylated RNA. Recombinant plasmids containing copies of polyadenylated RNA species were constructed using the G-C tailing procedure and clones of auxin-regulated sequences were detected by differential in situ hybridization with cDNA of polyadenylated RNA from auxin-treated or untreated hypocotyls. Although the 12 clones which were selected all contained different size inserts, and were therefore independent, 11 of these apparently hybridized to just two different RNA species. The rate constant of the auxin-sensitive abundant component of the untreated polyadenylated RNA/DNA hybridization was similar to that of the reaction between the two major groups of clones and untreated polyadenylated RNA. This indicates that these cloned sequences are homologous with that cDNA fraction. The twelfth clone is thought to be representative of a group of less abundnt auxin-regulated polyadenylated mRNA species which had been detected in an earlier analysis of the in vitro translation products of soybean hypocotyl RNA. Both the timing and the extent of the influence of auxin on the relative concentration of these cloned sequences are quite consistent with a close relationship between growth regulation by auxin and its effects on gene expression.


Crystallization and preliminary X-ray diffraction studies of the prototypal homologue of mitoNEET (Tth-NEET0026) from the extreme thermophile Thermus thermophilus HB8.
MitoNEET (a mammalian mitochondrial outer membrane protein) is a potential pharmacological and clinical target of the insulin-sensitizer pioglitazone. The thermophilic homologue of mitoNEET (TTHA0026) from Thermus thermophilus HB8 has been heterologously overproduced in Escherichia coli and purified as a water-soluble prototypal protein containing the mitoNEET-like [2Fe-2S] cluster. The resultant recombinant protein, named Tth-NEET0026, has been crystallized in its oxidized form by the hanging-drop vapour-diffusion method using 17%(w/v) polyethylene glycol 4000, 8.5%(v/v) 2-propanol, 15%(v/v) glycerol and 0.085 M HEPES-NaOH pH 7.2. The dark reddish crystals diffracted to 1.80 A resolution and belonged to the tetragonal space group P4(3)2(1)2, with unit-cell parameters a = 45.51, c = 84.26 A. The asymmetric unit contains one protein molecule.


Conditional Random Field (CRF)-Boosting: Constructing a Robust Online Hybrid Boosting Multiple Object Tracker Facilitated by CRF Learning.
Due to the reasonably acceptable performance of state-of-the-art object detectors, tracking-by-detection is a standard strategy for visual multi-object tracking (MOT). In particular, online MOT is more demanding due to its diverse applications in time-critical situations. A main issue of realizing online MOT is how to associate noisy object detection results on a new frame with previously being tracked objects. In this work, we propose a multi-object tracker method called CRF-boosting which utilizes a hybrid data association method based on online hybrid boosting facilitated by a conditional random field (CRF) for establishing online MOT. For data association, learned CRF is used to generate reliable low-level tracklets and then these are used as the input of the hybrid boosting. To do so, while existing data association methods based on boosting algorithms have the necessity of training data having ground truth information to improve robustness, CRF-boosting ensures sufficient robustness without such information due to the synergetic cascaded learning procedure. Further, a hierarchical feature association framework is adopted to further improve MOT accuracy. From experimental results on public datasets, we could conclude that the benefit of proposed hybrid approach compared to the other competitive MOT systems is noticeable.


Effects of ethanol on mature offspring of mice given ethanol during pregnancy.
Male offspring of mice maintained on isocaloric liquid diets containing either 20% ethanol or sucrose derived calories during pregnancy were tested on their ability to discriminate different ethanol doses as adults. They were trained to lever-press for a food reward on each of two levers in an operant chamber, and were then maintained on an FR20 reinforcement schedule. After response rates stabilized, ethanol discrimination training was initiated by reinforcing only responses made on the drug appropriate-lever after i.p. injections of ethanol (1.0 g/kg) or water. After learning to discriminate the 1.0 g dose, the animals' ability to discriminate doses of 0.25, 0.50, 0.75, 1.0, and 1.5 g/kg was assessed by determining the percentage of responses made on the drug lever during a 2-min test period. Compared to sucrose controls, mice exposed to alcohol in utero learned the lever response more slowly and were less responsive to injected ethanol as evidenced by a reduced effect of the drug on response rates and by a reduction in their ability to discriminate the presence of injected ethanol. The results indicate that prenatal ethanol exposure can have long term consequences which reduce the effects of ethanol in fully mature animals.


Dopamine and cognitive control: sex-by-genotype interactions influence the capacity to switch attention.
Cognitive performance in healthy persons varies widely between individuals. Sex differences in cognition are well reported, and there is an emerging body of evidence suggesting that the relationship between dopaminergic neurotransmission, implicated in many cognitive functions, is modulated by sex. Here, we examine the influence of sex and genetic variations along the dopaminergic pathway on aspects of cognitive control. A total of 415 healthy individuals, selected from an international consortium linked to Brain Research and Integrative Neuroscience Network (BRAINnet), were genotyped for two common and functional genetic variations of dopamine regulating genes: the catechol-O-methyltransferase [COMT] gene (rs4680) and the dopamine receptor D2 [DRD2] gene (rs6277). Cognitive measures were selected to explore sustained attention (using a continuous performance task), switching of attention (using a Trails B adaptation) and working memory (a visual computerised adaptation of digit span). While there were no main effects for genotype across any tasks, analyses revealed significant sex by genotype interactions for the capacity to switch attention. In relation to COMT, superior performance was noted in females with the Val/Val genotype and for DRD2, superior performance was seen for TT females and CC males. These findings highlight the importance of considering genetic variation in baseline dopamine levels in addition to sex, when considering the impact of dopamine on cognition in healthy populations. These findings also have important implications for the many neuropsychiatric disorders that implicate dopamine, cognitive changes and sex differences.


Ambulatory Blood Pressure Control in Children and Young Adults After Kidney Transplantation.
Ambulatory blood pressure (BP) monitoring (ABPM) is the preferred method to characterize BP status, and its use in kidney transplant recipients is increasing. Data on longitudinal ambulatory BP (ABP) trends in pediatric and young adult kidney transplant recipients are limited. Retrospective review of a large cohort of children and young adults following kidney transplantation and evaluation of their ABP status over time and its associations with any patient and clinical characteristics. Two hundred and two patients had baseline ABPM available for analysis, and 123 of them had a follow up (median time 2.3 years) ABPM. At the time of follow up, more patients were treated for hypertension (80% vs. 72%, P = 0.02), and less patients had ambulatory hypertension (36% vs. 54%, P = 0.005), uncontrolled or untreated, compared with baseline, with 45% of all patients classified as having controlled hypertension (compared to 26% at baseline, P = 0.002). Prevalence of ambulatory hypertension decreased only in patients who were less than 18 years old at baseline. High baseline mean 24-hour systolic BP was independently associated with persistent hypertension. In young kidney transplant recipients followed by ABPM, the prevalence of ambulatory hypertension decreases over time, mainly due to the increased number of patients with controlled hypertension.


The use of Bactroban for infection management in burn patients.
The paper summarizes experience with the application of Bactroban which was used for local antimicrobial treatment in 25 patients at the Burn Centre of the Department of Plastic Surgery, Prague. Various types of deep skin layers with mixed infection were chosen for this therapy, mostly due to Staphylococcus, even though a resistant Staphylococcus never occurred during the testing period. Bactroban was applied in cases where commonly used preparations had failed, and its therapeutical results can be considered positive.


[Determination of residual epichlorohydrin and sym-dichloroisopropyl alcohol in cationic etherified reagent by gas chromatography].
A method for the determination of the residual epichlorohydrin and sym-dichloroisopropyl alcohol in cationic etherified reagent by gas chromatography has been established. Methyl benzoate, which has high extraction efficiency for the two components, was used as extractant. With an HP-5 capillary column, the two components were baseline separated and they eluted before the extractant. The linear ranges achieved were 5 - 590 mg/kg for epichlorohydrin and 21 - 480 mg/kg for sym-dichloroisopropyl alcohol. The limits of detection were 1.2 mg/kg for epichlorohydrin and 2.2 mg/kg for sym-dichloroisopropyl alcohol. Recoveries for epichlorohydrin were 95.93% - 103.42% with relative standard deviations of 2.4% - 10.6% and those for sym-dichloroisopropyl alcohol were 98.54% - 107.40% with relative standard deviations of 6.6% -11.1%. The method is simple, fast, and convenient.


Impaired mitogen responses of the non-leukaemic B cells from patients with chronic lymphocytic leukaemia.
We investigated the immunological mechanism of the low level of circulating immunoglobulin and depressed primary and secondary responses of patients with chronic lymphocytic leukaemia (B-CLL) using purified non-leukaemic B cells in vitro. To assess the function of the non-leukaemic B cells we separated them from the much larger leukaemic population, which expresses the pan-T cell marker CD5, by immunoabsorption using anti-CD5 antibodies and Dynabeads. Immunoglobulin production was measured after the cells had been cultured with the B cell mitogens, pokeweed mitogen (PWM) and Staphylococcus aureus Cowan strain 1 (SAC). Autologous T cells that were found to function normally in our systems were added to cultures containing PWM. Non-leukaemic B cells from 15 B-CLL patients produced 539 ng/ml, immunoglobulin (mean value) with SAC and 162 ng/ml with PWM compared with 14,182 and 5513 ng/ml, respectively, from B cells from normal, age-matched control patients. Most of the immunoglobulin produced in the non-leukaemic B cell cultures carried the light chain associated with the leukaemic clone. We conclude that even at early stages in the disease (12 patients were Rai stage 0 patients) when the total serum immunoglobulin levels are still near normal, the B cells respond poorly to B cell mitogens.


Phenotypic heterogeneity associated with identical mutations in residue 870 of the androgen receptor.
Mutations in the androgen receptor (AR) gene result in an X-linked recessive form of male pseudohermaphroditism known as the androgen-insensitivity syndrome (AIS). The alterations most frequently observed are missense or nonsense point mutations in exons 4-8 of the AR gene that affect the steroid-binding domain of the receptor in subjects with various degrees of androgen resistance. Despite the increasing number of AR mutations identified, a reliable genotype-phenotype correlation has not been established and individuals with the same molecular defect may exhibit different phenotypes. Here, we studied a patients with an AIS characterized by bilateral gynecomastia, normal male external genitalia, and normal sperm counts. Exon-specific polymerase chain reaction, single-stranded conformational polymorphism, and sequencing analysis of the subject's AR gene were performed in addition to hormone-binding assays in skin fibroblasts from the patient. A point mutation at codon 870 of the AR, changing alanine to valine, was detected. As AR missense mutations changing alanine 870 to valine have been previously described in 3 unrelated patients showing severe AIS phenotypes, we conclude that phenotypic heterogeneity associated to identical mutations in the AR gene is probably due to individual functional differences in AR coregulator molecules.


Influence of intraoperative positive end-expiratory pressure level on pulmonary complications in emergency major trauma surgery.
Pulmonary complications have a major impact on the morbidity and mortality of critically ill patients with multiple trauma. Intraoperative protective ventilation with low tidal volume may prevent lung injury and infection, whereas the role of positive end-expiratory pressure (PEEP) levels is unclear. The aim of this study was to evaluate the influence of different intraoperative PEEP levels on incidence of pulmonary complications after emergency trauma surgery. We retrospectively analysed data of multiple trauma patients who underwent emergency surgery within 24 h after injury in our level I trauma centre (n = 86). On the basis of their intraoperative PEEP level, patients were divided into a low PEEP group with a PEEP of < 8 mbar and a high PEEP group with a PEEP of 8 mbar or higher. Besides differences in body mass index and preoperative oxygenation, there were no differences in patients' baseline data. There was a significant difference between incidence of pneumonia within 7 days after trauma surgery, with an incidence 26.7% in the low PEEP group and 7.3% in the high PEEP group (p = 0.02). The low PEEP group had higher pulmonary infection scores at days 3 and 5 after surgery. Oxygenation was better in the higher PEEP group postoperatively. There was no difference with respect to the incidence of acute respiratory distress syndrome, the mortality up until hospital discharge or haemodynamic parameters between groups. Higher PEEP levels were associated with perioperative improvement of oxygenation and a lower incidence of pneumonia, without impairment of haemodynamics. Additional studies should be initiated to confirm these observations.


Chlorhexidine spray effects on plaque accumulation in developmentally disabled patients.
Chlorhexidine gluconate has been studied as an adjunct to standard oral hygiene techniques in developmentally disabled patients in reducing plaque accumulation. The purpose of this double blind crossover study was to determine the efficacy of two concentrations of chlorhexidine spray (0.12% and 0.06%) on plaque accumulation in developmentally disabled patients. Informed consent was obtained for 8 developmentally disabled patients 12 to 33 years of age. Patients were randomly assigned to one of two groups of four each. One group received twice daily 0.12% chlorhexidine spray while the other received the 0.06% chlorhexidine spray for 30 days, followed by twice daily chlorhexidine spray (0.12% and 0.06%) alternating groups, for a period of 30 days. Final plaque indices were determined. Normal oral hygiene practices continued throughout the study. Significant plaque reduction occurred with the 0.12% and 0.06% spray solutions. No significant differences occurred between the two concentrations. The results of this study suggest that twice daily chlorhexidine spray at a concentration of 0.06% is as effective in reducing plaque accumulation as a 0.12% chlorhexidine concentration.


Ethical issues in publishing in predatory journals.
Predatory journals, or journals that charge an article processing charge (APC) to authors, yet do not have the hallmarks of legitimate scholarly journals such as peer review and editing, Editorial Boards, editorial offices, and other editorial standards, pose a number of new ethical issues in journal publishing. This paper discusses ethical issues around predatory journals and publishing in them. These issues include misrepresentation; lack of editorial and publishing standards and practices; academic deception; research and funding wasted; lack of archived content; and undermining confidence in research literature. It is important that the scholarly community, including authors, institutions, editors, and publishers, support the legitimate scholarly research enterprise, and avoid supporting predatory journals by not publishing in them, serving as their editors or on the Editorial Boards, or permitting faculty to knowingly publish in them without consequences.


Amantadine-associated corneal edema treated with descemet's stripping automated endothelial keratoplasty.
The authors report the successful use of Descemet's stripping automated endothelial keratoplasty (DSAEK) to treat a 45-year-old woman with amantadine-associated corneal edema. Discontinuation of the medication and treatment with corticosteroids did not result in resolution of the edema. The patient underwent sequential phakic DSAEK in both eyes with significant anatomic, visual, and functional improvement. Histopathologic analysis of Descemet's membrane by light microscopy revealed a paucity of endothelial cells. This case highlights the importance of considering amantadine toxicity in the differential diagnosis of corneal edema without an identifiable ocular cause and suggests the utility of DSAEK in the treatment of this rare condition.


Effect of ciprofloxacin on killing of Actinobacillus actinomycetemcomitans by polymorphonuclear leukocytes.
Actinobacillus actinomycetemcomitans, a pathogen associated with aggressive periodontitis, resists phagocytic killing by polymorphonuclear leukocytes (PMNs). It is susceptible to ciprofloxacin, which PMNs actively accumulate. This study tested the hypothesis that ciprofloxacin-loaded PMNs are more effective at killing A. actinomycetemcomitans than control PMNs. Isolated human PMNs were loaded by brief incubation with 0.5 microg of ciprofloxacin/ml. Opsonized bacteria (ATCC 43718) were incubated at 37 degrees C with control and ciprofloxacin-loaded PMNs and in the presence and absence of 0.5 microg of ciprofloxacin/ml. When assayed at bacteria-to-PMN ratios of 30:1 and 90:1, ciprofloxacin-loaded PMNs killed significantly more bacteria and achieved significantly shorter half times for killing than control PMNs (P < 0.05; Tukey's test). At ratios of 3:1 and 10:1, these differences were not significant.


Biosynthesis of archaeal membrane ether lipids.
A vital function of the cell membrane in all living organism is to maintain the membrane permeability barrier and fluidity. The composition of the phospholipid bilayer is distinct in archaea when compared to bacteria and eukarya. In archaea, isoprenoid hydrocarbon side chains are linked via an ether bond to the sn-glycerol-1-phosphate backbone. In bacteria and eukarya on the other hand, fatty acid side chains are linked via an ester bond to the sn-glycerol-3-phosphate backbone. The polar head groups are globally shared in the three domains of life. The unique membrane lipids of archaea have been implicated not only in the survival and adaptation of the organisms to extreme environments but also to form the basis of the membrane composition of the last universal common ancestor (LUCA). In nature, a diverse range of archaeal lipids is found, the most common are the diether (or archaeol) and the tetraether (or caldarchaeol) lipids that form a monolayer. Variations in chain length, cyclization and other modifications lead to diversification of these lipids. The biosynthesis of these lipids is not yet well understood however progress in the last decade has led to a comprehensive understanding of the biosynthesis of archaeol. This review describes the current knowledge of the biosynthetic pathway of archaeal ether lipids; insights on the stability and robustness of archaeal lipid membranes; and evolutionary aspects of the lipid divide and the LUCA. It examines recent advances made in the field of pathway reconstruction in bacteria.


A question of belonging: race, social fit, and achievement.
Stigmatization can give rise to belonging uncertainty. In this state, people are sensitive to information diagnostic of the quality of their social connections. Two experiments tested how belonging uncertainty undermines the motivation and achievement of people whose group is negatively characterized in academic settings. In Experiment 1, students were led to believe that they might have few friends in an intellectual domain. Whereas White students were unaffected, Black students (stigmatized in academics) displayed a drop in their sense of belonging and potential. In Experiment 2, an intervention that mitigated doubts about social belonging in college raised the academic achievement (e.g., college grades) of Black students but not of White students. Implications for theories of achievement motivation and intervention are discussed.


[Status of intra-aortic balloon pump application and related factors in Chinese patients with acute myocardial infarction].
Objective: To investigate the current status of application of intra-aortic balloon pump(IABP) and analyze the factors which might impact the application of IABP in patients with acute myocardial infarction complicated with cardiac shock in China. Methods: In China Acute Myocardial Infarction (CAMI) Registry,a nationwide, multicenter(107 hospitals), prospective study, 26 592 patients with acute myocardial infarction were enrolled consecutively between January 1, 2013 and September 30, 2014. After excluding of 30 cases due to missing important data,26 562 cases were analyzed.The application status of IABP was analyzed and multivariate logistic regression analysis was performed to determine the factors related to IABP application. Results: A total of 785(3.0%) patients with acute myocardial infarction received IABP implantation, and 381(49.9%) patients belonged to preventive application of IABP before primary percutaneous coronary intervention,and 6(0.8%) patients were complicated with mechanical complications.There were 118(15.0%) patients with cardiac shock received IABP implantation, which accounted for 12.0%(118/984) of all patients with cardiac shock.Multivariate logistic regression analysis showed that the clinical independent factors of the decision of IABP insertion included dyslipidemia (OR=2.858, 95%CI 1.397-5.846, P=0.004),left ventricular ejection fraction (OR=0.977, 95%CI 0.961-0.994, P=0.009),usage of dopamine (OR=2.817, 95%CI 1.495-5.308, P=0.001), left main disease (OR=2.817, 95%CI 1.495-5.308, P=0.001), GRACE score (OR=1.006, 95%CI 1.000-1.011, P=0.034), receiving primary percutaneous coronary intervention (OR=4.508, 95%CI 1.673-12.146, P=0.003), teritiary hospitals (OR=2.562, 95%CI 1.498-4.384, P=0.001), and higher education of the patients (OR=2.183, 95%CI 1.056-4.509, P=0.016). Conclusions: Among the Chinese acute myocardial infarction patients who received IABP implantation, nearly half application of IABP are preventive implantation before primary percutaneous coronary intervention. Only a few patients complicated with cardiac shock received IABP insertion. The clinical conditions, grade of hospitals, degree of education impact the decision of IABP insertion for the patients with acute myocardial infarction. Clinical Trial Registry National Institutes of Health, NCT018746.


Atherosclerosis and autoimmunity.
The possible involvement of immunological mechanisms in the pathogenesis of atherosclerosis has been suggested intermittently since the early 1970s. Both humoral and cellular mechanisms have been proposed to participate in the onset and/or progression of atheromatous lesions, but the theories postulating the involvement of autoantibodies and immune complexes have met with considerable experimental support. Modified lipoproteins, particularly different forms of oxidized LDL, have been reported to elicit humoral immune responses in both experimental animals and humans. Oxidized LDL has been demonstrated in atheromatous lesions, anti-oxidized LDL antibodies have been detected in circulation and in atheromatous plaques, and immune complexes formed with LDL and anti-LDL have been isolated from the serum of patients with manifestations of atherosclerosis. In addition, in vitro-formed LDL-IC and IC isolated from patients have been demonstrated to cause intracellular accumulation of cholesteryl esters (CE) in human macrophages and fibroblasts. The accumulation of CE in macrophages exposed to LDL-IC is unique to this type of IC and is associated with a paradoxical overexpression of the native LDL receptor and with increase synthesis and release of interleukin 1 and TNF-alpha. The release of these cytokines in the subendothelial space may have a significant role in promoting the interaction of endothelial cells with mononuclear cells, causing endothelial cell damage directly or indirectly, and also in inducing smooth muscle cell proliferation. Thus, several lines of evidence suggest that humoral autoimmunity may play a significant role in the pathogenesis of atherosclerosis.


Bacillus halodurans OapB forms a high-affinity complex with the P13 region of the noncoding RNA OLE.
Noncoding RNAs (ncRNAs) longer than 200 nucleotides are rare in bacteria, likely because bacterial genomes are under strong evolutionary pressures to maintain a small genome size. Of the long ncRNAs unique to bacteria, the OLE (ornate, large, extremophilic) RNA class is among the largest and most structurally complex. OLE RNAs form a ribonucleoprotein (RNP) complex by partnering with at least two proteins, OapA and OapB, that directly bind OLE RNA. The biochemical functions of the OLE RNP complex remain unknown, but are required for proper adaptation to certain environmental stresses, such as cold temperatures, short chain alcohols, and high magnesium concentrations. In the current study, we used electrophoretic mobility shift assays to examine the binding of OLE RNA fragments by OapB and found that OapB recognizes a small subregion of OLE RNA, including stem P13, with a dissociation constant (KD ) of ∼700 pm Analyses with mutated RNA constructs, and the application of in vitro selection, revealed that strong binding of OLE RNA by OapB requires a stem containing a precisely located single-nucleotide bulge and a GNRA tetraloop. Although the vast majority of bacteria with the ole gene also have the oapB gene, there are many whose genomes contain oapB but lack ole, suggesting that OapB has other RNA partners in some species that might exhibit similar structural features.


How moist heat kills spores of Bacillus subtilis.
Populations of Bacillus subtilis spores in which 90 to 99.9% of the spores had been killed by moist heat gave only two fractions on equilibrium density gradient centrifugation: a fraction comprised of less dense spores that had lost their dipicolinic acid (DPA), undergone significant protein denaturation, and were all dead and a fraction with the same higher density as that of unheated spores. The latter fraction from heat-killed spore populations retained all of its DPA, but >/=98% of the spores could be dead. The dead spores that retained DPA germinated relatively normally with nutrient and nonnutrient germinants, but the outgrowth of these germinated spores was significantly compromised, perhaps because they had suffered damage to some proteins such that metabolic activity during outgrowth was greatly decreased. These results indicate that DPA release takes place well after spore killing by moist heat and that DPA release during moist-heat treatment is an all-or-nothing phenomenon; these findings also suggest that damage to one or more key spore proteins causes spore killing by moist heat.


Mitogenic stimulation accelerates influenza-induced mortality by increasing susceptibility of alveolar type II cells to infection.
Development of pneumonia is the most lethal consequence of influenza, increasing mortality more than 50-fold compared with uncomplicated infection. The spread of viral infection from conducting airways to the alveolar epithelium is therefore a pivotal event in influenza pathogenesis. We found that mitogenic stimulation with keratinocyte growth factor (KGF) markedly accelerated mortality after infectious challenge with influenza A virus (IAV). Coadministration of KGF with IAV markedly accelerated the spread of viral infection from the airways to alveoli compared with challenge with IAV alone, based on spatial and temporal analyses of viral nucleoprotein staining of lung tissue sections and dissociated lung cells. To better define the temporal relationship between KGF administration and susceptibility to IAV infection in vivo, we administered KGF 120, 48, 24, and 0 h before intrapulmonary IAV challenge and assessed the percentages of proliferating and IAV-infected, alveolar type II (AECII) cells in dispersed lung cell populations. Peak AECII infectivity coincided with the timing of KGF administration that also induced peak AECII proliferation. AECII from mice that were given intrapulmonary KGF before isolation and then infected with IAV ex vivo exhibited the same temporal pattern of proliferation and infectious susceptibility. KGF-induced increases in mortality, AECII proliferation, and enhanced IAV susceptibility were all reversed by pretreatment of the animals with the mTOR inhibitor rapamycin before mitogenic stimulation. Taken together, these data suggest mTOR signaling-dependent, mitogenic conditioning of AECII is a determinant of host susceptibility to infection with IAV.


The age distribution of type-specific high-risk human papillomavirus incidence in two population-based screening trials.
Age- and type-specific high-risk human papillomavirus (hrHPV) incidence estimates in screen-eligible women are relevant from a public health perspective because they provide an indication of the effect of vaccination on the occurrence of screen-positives in HPV-based screening. However, limited data from women over 25 years of age are available. In 24,105 hrHPV-negative women participating in Dutch (Population-Based Screening Study Amsterdam: POBASCAM) and Italian (New Technologies for Cervical Cancer: NTCC) population-based randomized controlled screening trials the age- and type-specific distribution of incident hrHPV infections detected at the next screening round was assessed. HPV types were grouped into vaccine (bivalent: HPV16/18; polyvalent HPV16/18/31/33/45/52/58) and nonvaccine types. The incidence of screen-detected hrHPV among women ages 29 to 56 years was 2.54% (95% confidence interval, 2.30-2.78) in POBASCAM and 2.77% (2.36-3.19) in NTCC. In both studies, the incidence of bivalent, polyvalent, and nonpolyvalent infections decreased with age (P < 0.0001). Among women with incident infection(s), vaccine-type positivity changed quadratically with age, in particular for the polyvalent vaccine (P values: POBASCAM: bivalent 0.264, polyvalent 0.038; NTCC bivalent 0.039, polyvalent 0.005). However, more than 20% and 50% of women with incident hrHPV were positive for bivalent and polyvalent vaccine types, respectively, in all ages in both studies. We observed decreasing age trends of hrHPV vaccine and nonvaccine type incidences and age-related differences in the vaccine-type positivity among women with incident infections. Most importantly, hrHPV infections continued to be detected in all ages and the contribution of vaccine types remained substantial. Our results indicate a considerable reduction of new hrHPV infections in vaccinated cohorts, ensuing revision of screening guidelines.


Glycan Node Analysis: A Bottom-up Approach to Glycomics.
Synthesized in a non-template-driven process by enzymes called glycosyltransferases, glycans are key players in various significant intra- and extracellular events. Many pathological conditions, notably cancer, affect gene expression, which can in turn deregulate the relative abundance and activity levels of glycoside hydrolase and glycosyltransferase enzymes. Unique aberrant whole glycans resulting from deregulated glycosyltransferase(s) are often present in trace quantities within complex biofluids, making their detection difficult and sometimes stochastic. However, with proper sample preparation, one of the oldest forms of mass spectrometry (gas chromatography-mass spectrometry, GC-MS) can routinely detect the collection of branch-point and linkage-specific monosaccharides ("glycan nodes") present in complex biofluids. Complementary to traditional top-down glycomics techniques, the approach discussed herein involves the collection and condensation of each constituent glycan node in a sample into a single independent analytical signal, which provides detailed structural and quantitative information about changes to the glycome as a whole and reveals potentially deregulated glycosyltransferases. Improvements to the permethylation and subsequent liquid/liquid extraction stages provided herein enhance reproducibility and overall yield by facilitating minimal exposure of permethylated glycans to alkaline aqueous conditions. Modifications to the acetylation stage further increase the extent of reaction and overall yield. Despite their reproducibility, the overall yields of N-acetylhexosamine (HexNAc) partially permethylated alditol acetates (PMAAs) are shown to be inherently lower than their expected theoretical value relative to hexose PMAAs. Calculating the ratio of the area under the extracted ion chromatogram (XIC) for each individual hexose PMAA (or HexNAc PMAA) to the sum of such XIC areas for all hexoses (or HexNAcs) provides a new normalization method that facilitates relative quantification of individual glycan nodes in a sample. Although presently constrained in terms of its absolute limits of detection, this method expedites the analysis of clinical biofluids and shows considerable promise as a complementary approach to traditional top-down glycomics.


Association of APOE gene polymorphism with lipid profile and coronary artery disease in Afro-Caribbeans.
Apolipoprotein E gene (APOE) polymorphism is associated with the lipid profile and cardio-vascular disease. However, these relationships vary between ethnic groups. We evaluated, for the first time in an Afro-Caribbean population, the distribution of APOE polymorphisms and their associations with coronary artery disease (CAD), the lipid profile and other cardio-metabolic risk factors. We studied 712 Afro-Caribbean subjects including 220 with documented CAD and 492 healthy subjects. TaqMan assays were performed to genotype rs7412 and rs429358, the two variants that determine the APOE alleles ε2, ε3 and ε4. The association between APOE genotype and the lipid profile was analysed by comparing ε2 carriers, ε3 homozygotes and ε4 carriers. The frequencies of ε2, ε3 and ε4 in the overall sample were 8%, 70% and 22%, respectively. CAD was not associated with APOE polymorphism. The total cholesterol level was higher in ε4 carriers compared with ε2 carriers: 5.07 vs 4.59 mmol/L (P = 0.016). The LDL-cholesterol level was lower in APOE ε2 carriers compared with ε3 homozygotes and ε4 carriers: 2.65 vs 3.03 and 3.17 mmol/L, respectively (p = 0.002). The total cholesterol/HDL-cholesterol and LDL-cholesterol/HDL-cholesterol ratios were similar in the three allelic groups. APOE polymorphism was not associated with diabetes, hypertension, waist circumference or body mass index. Our results indicate that APOE gene polymorphism is associated with the lipid profile but not with CAD in Afro-Caribbean people. This lack of association with CAD may be explained by the low atherogenic profile observed in ε4 carriers, which may warrant further investigation.


Characterization of Theileria equi genotypes in horses in Israel, the Palestinian Authority and Jordan.
Equine theileriosis caused by Theileria equi is endemic in the Middle East, where it causes a severe disease as well as widespread subclinical infection. The aim of this study was to evaluate the diversity of T. equi genotypes in Israel and the neighboring Palestinian Authority and Jordan. Blood samples from 355 horses from Israel, the Palestinian Authority and Jordan were tested for the prevalence of T. equi DNA. Two hundred and fourteen (60%) were found positive for T. equi infection by PCR. Of those, the 18S rRNA (1458bp) and the EMA-1 (745bp) genes of T. equi were sequenced from 15 horse samples that represent Israel's geographical distribution together with four samples from the Palestinian Authority and two from Jordan. The results were used for genotype characterization and phylogenetic analysis of T. equi in the equine population in Israel and its surroundings. Three 18S rRNA genotype clades were found in Israel (A, C and D) with clade D being the most prevalent and included all four isolates from the PA. In contrast, the EMA-1 gene showed little diversity with all sequences clustering in the same clade apart from one Jordanian sequence. Results suggest that although the Israeli horse population is small and relatively confined geographically, it is probable that the genetic variability, which was found among Israeli horses, is a result of introduction of horses from other countries. It also suggests that the EMA-1 gene is probably not a good target for the evaluation of variance in T. equi populations. Characterization of the different genotypes prevalent in a certain region is important in order to map out the intra-species sequence heterogeneity of the parasite, which is needed in order to develop new diagnostic tools and vaccines.


A homobifunctional rhodamine for labeling proteins with defined orientations of a fluorophore.
The synthesis and characterization of a bifunctional rhodamine dye bearing 2-(iodoacetamido)ethyl substituents on the 3'- and 6'-nitrogen atoms is described. Aspects of the conversion of chloroacetamides to iodoacetamides are discussed, including a remarkably mild dehalogenation of an aromatic haloacetamide in the presence of NaI and camphorsulfonic acid. The bifunctional rhodamine has been designed for two-site, 1:1 labeling of proteins that contain two suitably disposed cysteine residues and is intended to constrain the orientation of the rhodamine absorption and emission dipoles in a predictable relationship to the protein structure.


Post-stroke depression, antidepressant treatment and rehabilitation results. A case-control study.
The aim of this study was to assess the specific influence of poststroke depression (PSD) on both basal functional status and rehabilitation results. We performed a case-control study in 290 stroke inpatients, matched for age (+/-1 year) and onset admission interval (+/-3 days) and divided in two groups according to the presence (PSD+) or absence (PSD-) of PSD. All PSD+ patients were treated with antidepressants (AD), mainly with fluoxetine. PSD+ patients, despite similar severity of stroke, showed greater disability in coping with activities of daily living (ADL) on admission and greater disability both in ADL and mobility at discharge than PSD- patients. Although both groups exhibited similar average functional improvement during rehabilitation, PSD- patients were nearly twice as likely to show excellent recovery both on ADL and mobility as PSD+ patients (OR = 1.95, 95% CI = 1.01-3.75 and OR = 2.23, 95% CI = 1.14-4.35, respectively). All AD drugs improved depressive symptoms. Few relevant side effects were observed: fluoxetine was discontinued in 2 patients because of insomnia and in 2 patients because of nausea; paroxetine was stopped in 1 patient because of nausea and dry mouth. Our results confirm the unfavorable influence of PSD on functional outcome, despite pharmacological treatment.


Quantitative EEG: II. Frequency analysis and topographic mapping in clinical settings.
The results of many studies using quantitative EEG techniques in clinical settings have been published. Those reports are reviewed here, with emphasis on those that used EEG frequency analysis and topographic mapping. In cerebrovascular disease, these methods can confirm the existence of lesions that are too mild to show up on routine EEG or too mild or too early to show up on computed tomography. The results correlate well with cerebral blood flow studies. These EEG tests can be done continuously in an intensive care unit or operating room. However, exact localization ability is inferior to that seen using traditional neuroimaging tests. In epilepsy, quantitative EEG techniques have found subtle degrees of background EEG changes near epileptic foci. Other methods can quantify epileptic spikes in useful ways and can indicate which region is driving other regions during seizures. Quantification is also useful for measuring drug effects when drugs (such as thiopental) are given deliberately to provoke acute EEG changes. Other measurements of drug effects may become useful in the future. In patients with mass lesions and metabolic encephalopathies, quantitative EEG changes do occur, and some of these correlate with the clinical state. However, in the latter settings, the clinical advantages for patient care are not yet clear, especially in comparison to available neuro-imaging studies and other routine medical tests. For dementia, quantitative EEG techniques are being developed. Some of these tests are accurate in moderately or severely demented patients, but there is still poor accuracy for early or borderline cases. For dyslexia, schizophrenia, and depression, there is a considerable volume of research reports but still no consensus about how to use quantitative EEG tests for care of individual patients. These tests require substantial user expertise in EEG. At present, these tests should be viewed as adjunctive to traditional EEG testing: such routine EEG testing should serve as the foundation for any clinical use of quantitative EEG tools.


COVID-19, emergency remote teaching evaluation: the case of Indonesia.
The global crisis caused by the COVID-19 pandemic has challenged educational institutions worldwide to rapidly shift to an online mode of teaching. In this paper, we discuss the concept of emergency remote teaching (ERT), including its implementation and evaluation, in the context of higher education in Indonesia. The Context, Input, Process, and Product framework was used to evaluate the implementation of ERT based on the experiences of 45 faculty members and 82 students from seven universities and colleges in three provinces in Indonesia. This study revealed several points of view. First, the shift to the ERT process depends on various aspects: internal organizational resources (curriculum, staff development, and technology), and external challenges (lack of access to a fast, affordable, and reliable Internet connection and the socioeconomic problems of the participants). Second, the ERT learning design needs to be framed using three principles: simplicity, flexibility, and empathy. The schools/administrators understand that this is not a normal situation in which learning competency standards must be rigorously met. In a crisis, given the facts that show disparities in technology and Internet networks, curriculum fulfillment is not the sole issue; it is also important to care for and support learners during this difficult time. This study provides recommendations that will serve as input for future strategies and educational policies in Indonesia, and developing countries in general. Additionally, this study can also be used as a benchmark for evaluating learning in similar situations in other countries.


Sigma-1 receptors control immune-driven peripheral opioid analgesia during inflammation in mice.
Sigma-1 antagonism potentiates the antinociceptive effects of opioid drugs, so sigma-1 receptors constitute a biological brake to opioid drug-induced analgesia. The pathophysiological role of this process is unknown. We aimed to investigate whether sigma-1 antagonism reduces inflammatory pain through the disinhibition of the endogenous opioidergic system in mice. The selective sigma-1 antagonists BD-1063 and S1RA abolished mechanical and thermal hyperalgesia in mice with carrageenan-induced acute (3 h) inflammation. Sigma-1-mediated antihyperalgesia was reversed by the opioid antagonists naloxone and naloxone methiodide (a peripherally restricted naloxone analog) and by local administration at the inflamed site of monoclonal antibody 3-E7, which recognizes the pan-opioid sequence Tyr-Gly-Gly-Phe at the N terminus of most endogenous opioid peptides (EOPs). Neutrophils expressed pro-opiomelanocortin, the precursor of β-endorphin (a known EOP), and constituted the majority of the acute immune infiltrate. β-endorphin levels increased in the inflamed paw, and this increase and the antihyperalgesic effects of sigma-1 antagonism were abolished by reducing the neutrophil load with in vivo administration of an anti-Ly6G antibody. The opioid-dependent sigma-1 antihyperalgesic effects were preserved 5 d after carrageenan administration, where macrophages/monocytes were found to express pro-opiomelanocortin and to now constitute the majority of the immune infiltrate. These results suggest that immune cells harboring EOPs are needed for the antihyperalgesic effects of sigma-1 antagonism during inflammation. In conclusion, sigma-1 receptors curtail immune-driven peripheral opioid analgesia, and sigma-1 antagonism produces local opioid analgesia by enhancing the action of EOPs of immune origin, maximizing the analgesic potential of immune cells that naturally accumulate in painful inflamed areas.


A Wnt-TGFβ2 axis induces a fibrogenic program in muscle stem cells from dystrophic mice.
We have previously observed that Wnt signaling activates a fibrogenic program in adult muscle stem cells, called satellite cells, during aging. We genetically labeled satellite cells in a mouse model of Duchenne muscular dystrophy to follow their fate during the progression of the disease. We observed that a fraction of satellite cells had a reduced myogenic potential and showed enhanced expression of profibrotic genes compared to age-matched controls. By combining in vitro and in vivo results, we found that expression of transforming growth factor-β2 (TGFβ2) was induced in response to elevated canonical Wnt signaling in dystrophic muscles and that the resulting increase in TGFβ activity affected the behavior of satellite cells in an autocrine or paracrine fashion. Indeed, pharmacological inhibition of the TGFβ pathway in vivo reduced the fibrogenic characteristics of satellite cells. These studies shed new light on the cellular and molecular mechanisms responsible for stem cell dysfunction in dystrophic muscle and may contribute to the development of more effective and specific therapeutic approaches for the prevention of muscle fibrosis.


Growth among HIV-infected children receiving antiretroviral therapy in Dar es Salaam, Tanzania.
We assembled a prospective cohort of 3144 human immunodeficiency virus (HIV) infected children aged <15 years initiating antiretroviral therapy (ART) in Dar es Salaam, Tanzania. The prospective relationships of baseline covariates with growth were examined using linear regression models. ART led to improvement in mean weight-for-age (WAZ), height/length-for-age (HAZ) and weight-for-length or body mass index (WLZ/BMIZ) scores. However, normal HAZ values were not attained over an average follow-up of 17.2 months. After 6 months of ART, underweight (P < 0.001), low CD4 count or percent (P < 0.001), stavudine containing regimens (P = 0.05) and advanced WHO disease stage (P < 0.001) at ART initiation were associated with better WAZ scores. Age >5 years on the other hand was associated with less increase in WAZ score after 6 months of ART (P < 0.001). These findings suggest that although ART improved the growth of the HIV-infected children in Tanzania, adjunct nutritional interventions may be needed to ensure that the growth of these children is optimized to the greatest extent possible.


A case of ochronotic arthropathy treated with total hip arthroplasty.
Abstract We report a case of ochronotic arthropathy treated with total hip arthroplasty. The articular cartilage of the patient's femoral head and synovium displayed black pigmentation. Microscopically, the articular cartilage revealed reddish-brown pigmentation (particularly in deep cartilage), necrotic chondrocytes, and slight black pigmentation in the cytoplasm of chondrocytes. Electron microscopy revealed electron-dense material deposited predominantly along and between collagen fibers.


Hydrogel-Based Active Substance Release Systems for Cosmetology and Dermatology Application: A Review.
Hydrogels are playing an increasingly important role in medicine and pharmacy. Due to their favorable physicochemical properties, biocompatibility, and designed interaction with living surroundings, they seem to be one of the most promising groups of biomaterials. Hydrogel formulations from natural, semi, or synthetic polymeric materials have gained great attention in recent years for treating various dermatology maladies and for cosmetology procedures. The purpose of this review is to present a brief review on the basic concept of hydrogels, synthesis methods, relevant mechanisms, and applications in dermatology or cosmetology. This review discusses transdermal therapies and the recent advances that have occurred in the field.


Engineering of vascular grafts with genetically modified bone marrow mesenchymal stem cells on poly (propylene carbonate) graft.
Bone marrow mesenchymal stem cells (MSCs) have demonstrated their pluripotency to differentiate into different cell lineages and may be an alternative cell source for vascular tissue engineering. The objective of this study is to create small diameter vessels by seeding and culture of genetically modified MSCs onto a synthetic polymer scaffold produced by an electrospinning technique. A tubular scaffold (2 mm in diameter) with a microstructure of nonwoven fibers was produced by electrospinning of poly (propylene carbonate) (PPC). Rat MSCs obtained from bone marrow were expanded in culture and modified with vasculoprotective gene endothelial nitric oxide synthase (eNOS) or marker gene green fluorescent protein (GFP). These MSCs were seeded onto the electrospun fibrous grafts (internal diameter = 2 mm), and cultured in 5% CO(2) at 37 degrees C. The growth of MSCs in the scaffold was analyzed with scanning electron microscopy (SEM) and hematoxylin and eosin (H&E) staining. The gene transfer and transgenic gene expression were examined with fluorescence-activated cell sorting (FACS), immunochemical staining, reverse transcriptase-polymerase chain reaction (RT-PCR), and western blot. The production of nitric oxide (NO) by the engineered vessels was measured with an NO detection kit. Our data showed that the seeded cells integrated with the microfibers of the scaffold to form a three-dimensional cellular network, indicating a favorable interaction between this synthetic PPC scaffold with MSCs. High transduction efficiency was obtained with the use of concentrated retrovirus in the gene transfection of MSCs. The eNOS gene transcripts and protein were detected in the grafts seeded with eNOS-modified MSCs by RT-PCR and immunochemical staining. The amount of NO produced by grafts seeded with eNOS-modified MSCs was comparable to that produced by native blood vessels, and it was significantly higher than that in the grafts seeded with nonmodified MSCs. In summary, the vascular graft produced by culture of eNOS gene-modified MSCs onto the electrospun tubular scaffolds shows promising results in terms of function. The use of MSCs and therapeutic genes in tissue engineering of blood vessels could be helpful in improving vessel regeneration and patency.


Hemiptera phylogenomic resources: Tree-based orthology prediction and conserved exon identification.
High-throughput sequencing of transcriptomes and targeted genomic regions are advancing our knowledge of The Tree of Life. Building phylogenies with regions of the genome requires 1-to-1 orthologue resources of genes and noncoding loci. One organismal group that has received little attention in this area is the Hemiptera, the fifth largest insect order represented by ~103,590 named species. Here, we present a set of 3,872 Hemiptera 1-to-1 orthogroups based on tree-based orthology inference of eight Hemiptera species with publicly available genome sequences. We also estimate a set of 406 orthologous exons with similar mRNA splice sites that can be used for Sanger sequencing and develop enrichment probes for targeted genome sequencing for phylogenomic inference. We show this novel set of orthologues is informative at the protein, coding sequence and exon molecular levels and provides robust branch support in both gene tree-species tree methods and concatenated sequence phylogenies. In addition, we demonstrate the utility of these loci to resolve relationships in whiteflies, Bemisia tabaci, a large species complex with few phylogenomic resources. Last, we compare our Hemiptera phylogeny with previously published phylogenies and other orthologue databases, while providing suggestions on further improvement to this phylogenomic resource.


Role of inositol 1,4,5-trisphosphate receptors in apoptosis in DT40 lymphocytes.
The role of inositol 1,4,5-trisphosphate receptors (IP(3)R) in caspase-3 activation and cell death was investigated in DT40 chicken B-lymphocytes stably expressing various IP(3)R constructs. Both full-length type-I IP(3)R and a truncated construct corresponding to the caspase-3 cleaved "channel-only" fragment were able to support staurosporine (STS)-induced caspase-3 activation and cell death even when the IP(3)R construct harbored a mutation that inactivates the pore of the Ca(2+) channel (D2550A). However, a full-length wild-type IP(3)R did not promote caspase-3 activation when the 159-amino acid cytosol-exposed C-terminal tail was deleted. STS caused an increase in cytosolic free Ca(2+) in DT40 cells expressing wild-type or pore-dead IP(3)R mutants. However, in the latter case all the Ca(2+) increase originated from Ca(2+) entry across the plasma membrane. Caspase-3 activation of pore-dead DT40 cells was also more sensitive to extracellular Ca(2+) chelation when compared with wild-type cells. STS-mediated release of cytochrome c into the cytosol and mitochondrial membrane potential depolarization could also be observed in DT40 cells lacking IP(3)Rs or containing the pore-dead mutant. We conclude that nonfunctional IP(3)Rs can sustain apoptosis in DT40 lymphocytes, because they facilitate Ca(2+) entry mechanisms across the plasma membrane. Although the intrinsic ion-channel function of IP(3)Rs is dispensable for apoptosis induced by STS, the C-terminal tail of IP(3)Rs appears to be essential, possibly reflecting key protein-protein interactions with this domain.


Complex dielectric relaxation in supercooling and superpressing liquid-crystalline chiral isopentylcyanobiphenyl.
Results of broadband dielectric studies in glass-forming liquid crystalline chiral isopentylcyanobiphenyl (5(*)CB) are presented. Tests conducted as a function of temperature and pressure revealed the coexistence of glassy and critical properties. The latter are associated with the isotropic-cholesteric phase transition at T(I-Ch) approximately 250 K under atmospheric pressure. Dielectric loss curves in the isotropic liquid and in the cholesteric phase are clearly broadened on cooling and pressuring towards the glass transition. Although in the isotropic phase there is a single stretched loss curve, in the mesophase an additional relaxation process can be distinguished. The evolution of relaxation times is non-Arrhenius and can be portrayed by the Vogel-Fulcher-Tamman relation or its pressure counterpart. The glassy dynamics coexists with the critical-like behavior for the static dielectric permittivity and for the maxima of the dielectric loss curves. Their temperature and pressure dependences are associated with the critical exponent phi=1-alpha approximately 1/2, where alpha approximately 1/2 is the specific heat critical exponent. This behavior is associated with the continuous phase transition placed at DeltaT approximately 1.5 K below the clearing temperature for P=0.1 MPa. It has been found that 5(*)CB shows a unique pressure-temperature phase diagram. Pressure and temperature changes which begin in the isotropic liquid below at ca. T approximately 265 K always result in the transition to the cholesteric phase which can be supercooled or superpressed. For T>265 K the phase transition to another phase, presumably a solid one, always occurs. However, a cholesteric-solid phase border seems to exist only in isothermal pressure tests. It does not appear in the temperature studies.