Association between mothers' and fathers' depressive symptoms, sense of coherence and perception of their child's temperament in early parenthood in Sweden.
To examine whether there was any association between mothers' and fathers' post-partum depressive symptoms and sense of coherence and perception of their child's temperament. The hypotheses were that parents with depressive symptoms: 1) have more often a poor sense of coherence, and 2) perceive their child's temperament to be more difficult than parents without depressive symptoms. A total of 401 Swedish-speaking couples, who were the parents of children born through the years 2004-2006 in the northern part of the county of Västmanland, Sweden, were invited to participate in the study. The parents answered 3 questionnaires including: at inclusion of the study: demographic data (n = 393 couples); at 3 months: the Edinburgh Postnatal Depression Scale and the Sense of Coherence Scale (n = 308 couples); and at 18 months: the Infant Characteristics Questionnaire (n = 272 couples). Depressive symptoms measured at 3 months, were reported by 17.7% of mothers and 8.7% of fathers, and correlated significantly between mothers and fathers within couples (rho = 0.165, p = 0.003). Mothers and fathers with depressive symptoms had a poorer sense of coherence (p < 0.001, p < 0.001) and perceived their child's temperament as more difficult than mothers and fathers without depressive symptoms at 3 (p = 0.028, p < 0.001) and 18 months (p = 0.145, p = 0.012 respectively). Early parenthood has been studied thoroughly in mothers, but few studies have included fathers. Identifying problems in early parenthood could help predict later problems exhibited by the preschool child, which might be prevented by supportive programmes.


Workshop: hypertension and cardiovascular risk factors: role of the angiotensin II-nitric oxide interaction.
Vascular upregulation of nitric oxide (NO) is an adaptive response to increased blood pressure that may help in the prevention of end-organ damage. Differences in cardiovascular and renal morbidity and mortality in hypertensive patients may result, at least in part, from individual variations in endothelial function in response to the hemodynamic workload of hypertension. A functional feedback balance exists between both angiotensin (Ang) II and NO under normal conditions. The NO-Ang II imbalance may not explain all the vascular pathophysiology of hypertension, but it certainly appears to be an important component. In hypertension, salt sensitivity, whether primary (ie, certain populations in the United States and Japan) or secondary (ie, aging, type II diabetes), appears to be a marker of increased cardiovascular and renal risk that is often linked to a decreased bioactivity of NO. In diabetes and atherosclerosis, NO-dependent vascular relaxation is impaired and can be restored by decreasing the synthesis and/or blocking the action of Ang II. An understanding of the relations between hypertension, cardiovascular risk factors, end-organ damage, and the NO-Ang II axis leads one to believe that the combination of therapeutic agents capable of reinstating the homeostatic balance of these vasoactive molecules within the vessel wall would be most effective in preventing or arresting end-organ disease.


[The mechanism of action of trisportic acids on the carotene-synthesizing enzymes of the (-) strain of Blakeslea trispora].
Trisporic acids were found to increase the content of carotene, protein, and RNA in the (-)strain of Blakeslea trispora. Trisporic acids are inductors of the synthesis of carotenogenous enzymes as was found in experiments with the inhibitor of transcription (cycloheximide) and translation (actinomycin D). Trisporic acids stimulate the formation of neutral fractions of the sex hormone, the process being regulated presumably at the level of posttranscription control.


Cutting edge: Hematopoietic-derived APCs select regulatory T cells in thymus.
Recognition of self-peptide-MHC complexes by high-affinity TCRs and CD28 signaling are critical for the development of forkhead-winged helix box transcription factor 3(+) regulatory T cells (Tregs) in thymus. However, the type of APCs that are responsible for selecting Tregs has remained unclear. To dissect the role of hematopoietic-derived APCs (HCs) and thymic epithelial cells (TECs) in Treg selection, we constructed bone marrow chimeras with disrupted CD28/B7 signaling in the HC or TEC compartment and analyzed the generation of Tregs in the thymus. We found that both HCs and TECs were independently able to fully reconstitute the Treg population in the thymus of bone marrow chimeras. In addition, Treg selection requires the TCR signal and CD28 costimulation presented in cis on the same APC type in vivo. This study demonstrates a new role, to our knowledge, for HCs in the development of Tregs in thymus.


Relationships between enterococcal virulence and antimicrobial resistance.
Enterococci have become a vexing problem in clinical medicine because of their ability to infect patients who are typically receiving antibiotic therapy for unrelated underlying illness. Moreover, the infections have become extremely difficult to manage because of the accumulation of antibiotic resistances among enterococci. The ability of enterococci to cause disease is an intrinsic property of the organism or possibly subpopulations within enterococcal species. The probability of an infection's becoming established, however, is almost certainly in part a function of the enterococcal burden. By altering endogenous bacterial flora, antibiotic therapy promotes increased colonization by antibiotic-resistant organisms. Therefore, antibiotic resistance and intrinsic virulence both contribute to disease, but in separate and complementary ways. We review the virulence of enterococci, as distinct from the acquisition of antimicrobial resistance genes, and identify current gaps in our understanding of enterococcal virulence and the basis for disease.


Obesity and skin and soft tissue infections: how to optimize antimicrobial usage for prevention and treatment?
Skin and soft tissue infections (SSTIs) are prevalent in the obese population, with rising trend expected. Although numerous antibiotics are available for the prevention and treatment of SSTIs, their characterization in obese patients is not a regulatory mandate. Consequently, information that carries importance for optimizing the dosing regimen in the obese population may not be readily available. This review focuses on the most recent pharmacokinetic and pharmacodynamic data on this topic with attention to cefazolin for surgical prophylaxis as well as antibiotics that are active against methicillin-resistant Staphylococcus aureus (MRSA). Moreover, the implications for optimizing SSTIs prevention and treatment in the obese population will also be discussed. On the basis of pharmacokinetic/pharmacodynamic considerations, most studies found a perioperative prophylactic cefazolin regimen of 2 g to be reasonable in the case of obese patients undergoing cesarean delivery or bariatric surgery. There is general paucity of data regarding the pharmacokinetic/pharmacodynamic characteristics of antimicrobials active against MRSA in obese patients, especially for the target tissue. Therapeutic drug monitoring has been correlated with pharmacokinetic/pharmacodynamic optimization for vancomycin and teicoplanin, and should be used in these cases. There is more supportive evidence for the use of oxazolidinones (linezolid and tedizolid), daptomycin and lipoglycopeptides (telavancin, dalbavancin and oritavancin) in the management of SSTIs in this population. The pharmacokinetic/pharmacodynamic approach, which can be used as a basis or supplement to clinical trials, provides valuable data and decision-making tools for optimizing regimens used for both prevention and treatment of SSTIs in the obese population. Important pharmacokinetic/pharmacodynamic characteristics of antibiotics, such as the penetration into the subcutaneous tissue and the probability of reaching the pharmacodynamic, target dictate efficacy, and thus should be taken into account and further investigated.


Follicular Dendritic Cell Sarcoma of the Inflammatory Pseudotumor-like Variant Presenting as a Colonic Polyp.
Follicular dendritic cell (FDC) sarcoma is rare and is classified either as conventional type or inflammatory pseudotumor (IPT)-like variant. Extranodal presentation is uncommon and nearly all gastrointestinal FDC tumors are of the conventional type. IPT-like variant tumors occur almost exclusively in the liver and spleen and are consistently associated with Epstein-Barr virus (EBV). Here we report the case of a 78-year-old woman with an IPT-like FDC sarcoma presenting as a pedunculated colonic polyp. Histologically, scanty atypical ovoid to spindle cells were mixed with a background of florid lymphoplasmacytic infiltrate, which led to an initial misdiagnosis of pseudolymphoma. These atypical cells expressed CD21, CD23, CD35, and D2-40, and were positive for EBV by in situ hybridization, confirming the diagnosis. The patient was free of disease five months after polypectomy without adjuvant therapy. Although extremely rare, the differential diagnosis for colonic polyp should include FDC sarcoma to avoid an erroneous diagnosis. A review of the 24 cases of IPT-like FDC sarcoma reported in the literature reveal that this tumor occurs predominantly in females with a predilection for liver and spleen, and has a strong association with EBV.


Twin pregnancies following induction of ovulation: a literature review.
A literature review of the occurrence of multiple pregnancies associated with artificial induction of ovulation is reported. This report considers three treatment schedules: (1) clomiphene citrate; (2) human pituitary gonadotrophin with human chorionic gonadotrophin; and (3) human menopausal gonadotrophin with human chorionic gonadotrophin. The majority of the increase in twinning is related to hyperstimulation of the ovary by these medications, resulting in dizygotic twinning. The true incidence of twin pregnancy cannot be calculated because the vital statistics of all nations report live birth rates. Increased rates of fetal wastage, late abortion and prematurity associated with the occurrence of multiple pregnancies are overlooked by these statistics. The increased incidence of twinning appears to be related to the type and dosage of medication used, and the patient's underlying problem.


Immunoisolation of adult porcine islets for the treatment of diabetes mellitus. The use of photopolymerizable polyethylene glycol in the conformal coating of mass-isolated porcine islets.
Functional porcine islets, free of known pathogens, can serve as a source of insulin producing cells for the treatment of experimentally induced insulin dependent Diabetes Mellitus. Porcine islets can be conformally coated (microencapsulated) with a covalently linked, stable permselective membrane while maintaining islet viability and function. The PEG conformal coating is immunoprotective in a discordant xenograft animal model (porcine islets to rat).


Endocochlear potential and scala media oxygen tension during partial anoxia.
The endocochlear potential and the scala media oxygen tension were measured from the basal coil of the guinea pig cochlea during levels of partial respiratory anoxia. This was accomplished by simultaneously positioning two electrodes (for endocochlear potential and PO2) into the endolymph by a transbasilar membrane approach, in order to minimize blood capillary damage. The intracochlear pO2 was stable until the respiratory oxygen concentration was reduced to 16 per cent on the average. Thereafter, a further reduction of oxygen in the respiratory mix reduced both the endolymph pO2 and the endocochlear potential. The coupling between the endocochlear potential and endolymph pO2 is proportional over a 70 to 80 per cent reduction in endolymph pO2, and at this point the endocochlear potential is reduced by 50 per cent. For partial levels of respiratory oxygen that are not too low (e.g., > 12 per cent oxygen) it is possible to maintain partial levels of intracochlear pO2 for at least six minutes. During this time period the endocochlear potential is also held at a stable but reduced level.


Uncovertebral and facet joint dislocations in cervical articular pillar fractures: CT evaluation.
Computed tomographic (CT) scans and plain radiographs of 21 patients with surgically treated fractures or dislocations of the cervical spine were reviewed. CT effectively demonstrated the locked or perched facets and half of the 13 articular pillar fractures. The fracture lines through the articular pillar were difficult to detect in some cases or to distinguish from a facet joint in others. However, distraction of an adjacent uncovertebral or facet joint was demonstrated by CT in each case of articular pillar fracture or locked or perched facet. Therefore, CT demonstration of a distracted facet or uncovertebral joint is an indirect sign of an unstable fracture or of a dislocation that may be more readily recognized on plain radiographs or pluridirectional tomograms.


High‑dose folic acid improves endothelial function by increasing tetrahydrobiopterin and decreasing homocysteine levels.
The aim of this study was to investigate the effect of folic acid (FA) on tetrahydrobiopterin (BH4), neopterin, nitric oxide (NO) and homocysteine (Hcy) levels in endothelial cells. Human umbilical vein endothelial cells (HUVECs) were cultured in vitro in the presence or absence of Hcy. The effect of various doses of FA on Hcy, BH4, neopterin and NO concentrations in HUVECs was then assessed. In the 5 and 10 nmol/l FA treatment groups, FA was found to significantly increase the levels of BH4 (10.56±3.86 and 11.23±2.1919 pmol/g vs 6.32+2.87 nmol/g; P<0.05 vs. control) and NO production (37.86±12.34 nmol/l, 38.45±11.23 nmol/l vs 26.21±9.24 nmol/l; P<0.001 vs. paired Hcy group), but reduce the levels of Hcy (132.87±29.67 and 140.87±26.76 nmol/l vs. 165.23±30.56 nmol/l; P<0.05 vs. Hcy group). No significant differences were observed in neopterin levels among the different groups of HUVECs. In conclusion, high doses of FA may be capable of protecting endothelial cells through reducing levels of Hcy and increasing BH4 and NO production.


Modulatory neurotransmitter systems and behavior: towards zebrafish models of neurodegenerative diseases.
The modulatory aminergic neurotransmitters are involved in practically all important physiological systems in the brain, and many of them are also involved in human central nervous system diseases, including Parkinson's disease, schizophrenia, Alzheimer's disease, and depression. The zebrafish brain aminergic systems share many structural properties with the mammalian systems. The noradrenergic, serotonergic, and histaminergic systems are highly similar. The dopaminergic systems also show similarities with the major difference being the lack of dopaminergic neurons in zebrafish mesencephalon. Development of automated quantitative behavioral analysis methods for zebrafish and imaging systems of complete brain neurotransmitter networks have enabled comprehensive studies on these systems in normal and pathological conditions. It is possible to visualize complete neurotransmitter systems in the whole zebrafish brain at an age when the fish already displays all major vital behaviors except reproduction. Alterations of brain dopaminergic systems with MPTP, the neurotoxin that in humans and rodents induces Parkinson's disease, induces both changes in zebrafish dopaminergic system and quantifiable abnormalities in motor behavior. Chemically-induced brain histamine deficiency causes an identifiable alteration in histaminergic neurons and terminal networks, and a clear change in swimming behavior and long-term memory. Combining the imaging techniques and behavioral methods with zebrafish genetics is likely to help reveal how the modulatory transmitter systems interact to produce important behaviors, and how they are regulated in pathophysiological conditions and diseases.


Mechanisms governing the reactivation-dependent destabilization of memories and their role in extinction.
The extinction of learned associations has traditionally been considered to involve new learning, which competes with the original memory for control over behavior. However, a recent resurgence of interest in reactivation-dependent amnesia has revealed that the retrieval of fear-related memory (with what is essentially a brief extinction session) can result in its destabilization. This review discusses some of the cellular and molecular mechanisms that are involved in the destabilization of a memory following its reactivation and/or extinction, and investigates the evidence that extinction may involve both new learning as well as a partial destabilization-induced erasure of the original memory trace.


Relationship of Early Vitamin D Concentrations and Gestational Diabetes Mellitus in Indian Pregnant Women.
Background: A high prevalence of vitamin D deficiency exists in pregnant Indian women (~90%). Increasing evidence suggests that vitamin D could play a pivotal role in maintaining normal glucose homeostasis. We aimed to determine the association between maternal vitamin D concentrations in early pregnancy and the risk of gestational diabetes mellitus (GDM). Methods: A prospective observational study was conducted on healthy pregnant women (n = 392) attending routine antenatal care at St. John's Medical College Hospital, Bangalore recruited at ~12 weeks of gestation. At baseline, details on socio-economic status, obstetric history, dietary intakes, and anthropometry were collected. Venous plasma total vitamin D concentration was assessed using tandem liquid chromatography mass spectrophotometry (LC-MS/MS). Oral glucose tolerance test (OGTT) at recruitment, followed by glucose tolerance test (GTT) at mid-pregnancy was conducted. GDM was diagnosed and confirmed using the International Association of Diabetes and Pregnancy Study Groups (IADPSG) classification. Univariate and adjusted logistic regression models were used to evaluate the associations between total vitamin D concentrations at enrollment with GDM. Results: Of the cohort, 10.2% were diagnosed as GDM. Women with GDM were older (26 vs. 24 years) and heavier (51.6 vs. 51.2 kg) compared to the rest. A higher prevalence of GDM was observed among women with 1st trimester plasma total vitamin D in the lowest quartile (≤23.6 nmol/L) compared to the subjects in the other three quartiles (16.1 vs. 8.6%, p = 0.033). Adjusted multivariable regression analysis showed that women in the lowest quartile of plasma total vitamin D had twice the odds of GDM compared to women belonging to the remaining quartiles [OR = 2.32 (95%CI: 1.10, 4.91), p = 0.028]. Conclusions: Low plasma total vitamin D concentrations in early pregnancy may be associated with a higher risk of GDM.


Effects of ovarian steroids and pregnancy on adrenergic nerves of uterus and oviduct.
This review concerns the influence of ovarian steroids and of pregnancy on norepinephrine (NE) metabolism in the adrenergic neurons of the female reproductive tract and speculates on the physiological consequences of this influence. Estrogen and progesterone affect not only the NE content of these nerves but also the turnover of NE, the activity of its synthetic enzyme, and releases of NE from nerve terminals. During pregnancy additional factors including stretch-induced hypertrophy come into play and cause degeneration of the nerves in the uterine corpus. This degeneration makes the muscle supersensitive to NE and may also induce morphological changes in the muscle cell membrane. As a result there may be a withdrawal of neural inhibitory influences on the corpus, allowing spontaneous myogenic contractions to intensify. Although the physiological significance of the steroid-transmitter interactions are still unclear, these nerves per se are of interest because they represent a model system for the study of neuroendocrine regulation in the peripheral nervous system.


Iconicity in the speech of children and adults.
Iconicity - the correspondence between form and meaning - may help young children learn to use new words. Early-learned words are higher in iconicity than later learned words. However, it remains unclear what role iconicity may play in actual language use. Here, we ask whether iconicity relates not just to the age at which words are acquired, but also to how frequently children and adults use the words in their speech. If iconicity serves to bootstrap word learning, then we would expect that children should say highly iconic words more frequently than less iconic words, especially early in development. We would also expect adults to use iconic words more often when speaking to children than to other adults. We examined the relationship between frequency and iconicity for approximately 2000 English words. Replicating previous findings, we found that more iconic words are learned earlier. Moreover, we found that more iconic words tend to be used more by younger children, and adults use more iconic words when speaking to children than to other adults. Together, our results show that young children not only learn words rated high in iconicity earlier than words low in iconicity, but they also produce these words more frequently in conversation - a pattern that is reciprocated by adults when speaking with children. Thus, the earliest conversations of children are relatively higher in iconicity, suggesting that this iconicity scaffolds the production and comprehension of spoken language during early development.


Bone Marrow Neutrophils of Multiple Myeloma Patients Exhibit Myeloid-Derived Suppressor Cell Activity.
Activated normal density granulocytes (NDGs) can suppress T-cell responses in a similar way as myeloid-derived suppressor cells (MDSCs). In this study, we tested the hypothesis that NDGs from blood and bone marrow of multiple myeloma (MM) patients have the ability to suppress T-cells, as MDSC. MM is an incurable plasma cell malignancy of the bone marrow. Like most malignancies, myeloma cells alter its microenvironment to promote tumor growth, including inhibition of the immune system. We found that MM NDG from the bone marrow suppressed proliferation of T-cells, in contrast to healthy donors. The inhibitory effect could not be explained by changed levels of mature or immature NDG in the bone marrow. Moreover, NDG isolated from the blood of both myeloma patients and healthy individuals could inhibit T-cell proliferation and IFN-γ production. On the contrary to previous studies, blood NDGs did not have to be preactivated to mediate suppressive effects. Instead, they became activated during coculture, indicating that contact with activated T-cells is important for their ability to regulate T-cells. The inhibitory effect was dependent on the production of reactive oxygen species and could be reverted by the addition of its inhibitor, catalase. Our findings suggest that blood NDGs from MM patients are suppressive, but no more than NDGs from healthy donors. However, only bone marrow NDG from MM patients exhibited MDSC function. This MDSC-like suppression mediated by bone marrow NDG could be important for the growth of malignant plasma cells in MM patients.


Older Patients' Competence, Preferences, and Attitudes Toward Digital Technology Use: Explorative Study.
Malnutrition is prevalent in older patients, which is associated with severe consequences such as a decline in functional status, increased risk of readmission, and increased mortality. A tablet-based eHealth solution (Food'n'Go) was recently developed and introduced at our clinic to support older patients' involvement in nutritional interventions during their hospitalization, thereby enhancing their awareness and motivation for choosing the right food to obtain sufficient calorie and protein intake. To reap the full benefits from the eHealth solution, the technology should be introduced and accompanied by support that targets the end users' competence level and needs. In this study, we aimed to explore older patients' readiness (ie, competence, preferences, and attitudes) toward the use of information and communication technology (ICT), and to identify the factors that may act as barriers or facilitators for their engagement with health technology. A descriptive and explorative study was performed using triangulation of data derived from semistructured interviews and questionnaires (based on the Readiness and Enablement Index for Health Technology [READHY] instrument). Older hospitalized patients (age ≥65 years; N=25) were included from two hospitals in Denmark. The majority (16/25, 64%) of the older patients (median age 81 years) were users of ICT. The qualitative findings revealed that their experiences of benefits related to the use of ICT facilitated usage. Barriers for use of ICT were health-related challenges, limited digital literacy, and low self-efficacy related to ICT use due to age-related prejudices by their relatives and themselves. The qualitative findings were also reflected in the low median scores on the eHealth Literacy Questionnaire (eHLQ) READHY scales within dimensions addressing the user's knowledge and skills (eHLQ1:1.8; eHLQ3: 2.0), and the user experience (eHLQ6: 2.0; eHLQ7: 1.5). Older patients are potential users of ICT, but experience a variety of barriers for using eHealth. When introducing older patients to eHealth, it is important to emphasize the possible benefits, and to offer support targeting their knowledge, skills, and motivation.


Dietary Bioactive Compounds: Implications for Oxidative Stress and Inflammation.
Nowadays, it has been amply demonstrated how an appropriate diet and lifestyle are essential for preserving wellbeing and preventing illnesses [...].


Dynamic changes in nucleus accumbens dopamine efflux during the Coolidge effect in male rats.
The Coolidge effect describes the reinitiation of sexual behavior in a "sexually satiated" animal in response to a novel receptive mate. Given the role of the mesolimbic dopamine (DA) system in the initiation and maintenance of motivated behavior, microdialysis was used to monitor nucleus accumbens (NAC) DA transmission during copulation, sexual satiety, and the reinitiation of sexual behavior. In agreement with earlier reports, the presentation of an estrous female behind a screen and copulation were associated with significant increases in NAC DA efflux. Return of NAC DA concentrations to baseline values coincided with a period of sexual satiety, although concentrations of the DA metabolites, dihydroxyphenylacetic acid and homovanillic acid, remained elevated. The presentation of a novel receptive female behind a screen resulted in a slight increase in NAC DA, which was augmented significantly during renewed copulation with the novel female. The present data suggest that the stimulus properties of a novel receptive female may serve to increase NAC DA transmission in a sexually satiated male rat, and this, in turn, may be related to the reinitiation of sexual behavior.


The factors affecting the institutionalisation of two policy units in Burkina Faso's health system: a case study.
This paper is one of three linked studies that attempts to understand the process of institutionalisation of policy units within Burkina Faso's health system. It examines the relationships between the existence of an institutional framework, data production capacity and other resource availability in the institutionalisation of policy units in health systems. It therefore contributes to our understanding of the dynamics linking the key drivers and indicators of institutionalisation. Additionally, it examines how factors within the managerial setting, including workplace environment, and budgetary and human resource availability, may influence the institutionalisation process. The study used an explanatory qualitative case study approach, examining two policy units in Burkina Faso's Ministry of Health, the first of which had been institutionalised successfully and the other less so. Data were collected from key policymakers, including 13 connected with the first policy unit and 10 with the second, plus two funders. We also conducted a documentary analysis of the National Program for Health Development, two mid-term strategic plans, 230 action plans, eight Ministry of Health state budgets, eight Ministry of Health annual statistics reports, 16 policy unit budgets and published literature. The framework within which the government gave the policy unit its mandate and policy focus had the strongest effect on the institutionalisation process. Institutionalisation depended on political will, in both the host government and any donors, and the priority given to the policy unit's focus. It was also affected by the leadership of the policy unit managers. These factors were influenced by human resource capacity, and our findings suggest that, for successful institutionalisation in Burkina Faso's health system, policy units need to be given sufficient human resources to achieve their objectives. Policy units' institutionalisation in Burkina Faso's health system depend on the leadership of the unit managers to implement relevant activities, mobilise funding, and recruit and maintain enough human resources, as well as the mandate given by the government.


Application of a Landmark-Based Method for Acoustic Analysis of Dysphonic Speech.
Speakers with dysphonia often report difficulty with maintaining intelligibility in noisy environments; however, there is no objective method for characterizing this difficulty. Landmark-based analysis is a linguistically-motived, knowledge-based speech analysis technique, which may serve as the basis of acoustic tool for describing the intelligibility deficit. As the first step toward development of such a tool, this study examined whether Landmark-based analysis could describe acoustic differences between normal and dysphonic speech. The recordings subjected to the Landmark-based analysis were the first sentence of the Rainbow Passage from 33 speakers with normal voice and 36 speakers with dysphonia. These recordings were selected from the Kay Elemetrics Database of Disordered Voice. The between-group difference was evaluated based on counts of certain Landmarks (LM). The average counts of all LMs were significantly greater in normal speech, t(66.85) = 2.36, P = 0.02. When the group-difference was examined for each LM, dysphonic speech had more [g] and [b] LMs and fewer [s] LMs than normal speech (P < 0.01 for all cases). A classification tree model identified [+s] and [+b] LMs are the primary predictors for the dysphonic speech. The model's misclassification rate was 7.24%. This preliminary investigation demonstrates that LM-based analysis is capable of differentiating dysphonic speech from normal speech. This encouraging result rationalizes future examinations of LM analysis in other areas of interest. For example, LM-based measures could conceivably be used as to quantify general intelligibility, and/or provide insight into underlying mechanisms of intelligibility deficits.


Crystal and molecular structure of adenosine 3',5'-cyclic phosphate.
The structure of adenosine 3',5'-cyclic phosphate has been determined by single-crystal x-ray diffraction. The two molecules in the asymmetric unit show different conformation about the glycosidic bond, while other structural details are essentially the same. The furanose rings are puckered with the C(4') atom out of the best four-atom plane. The bond lengths and angles appear to be normal.


Testosterone treatment of HSDD in naturally menopausal women: the ADORE study.
To evaluate the efficacy and safety of a transdermal testosterone patch (TTP, 300 microg/day) in naturally menopausal women with hypoactive sexual desire disorder (HSDD). A total of 272 naturally menopausal women, predominantly not using hormone therapy, were randomized in this 6-month, placebo-controlled, double-blind, multicenter study to receive twice weekly either TTP or an identical placebo. Efficacy endpoints measured were the 4-week frequency of satisfying sexual episodes (SSE) using the Sexual Activity Log, the sexual desire domain of the Profile of Female Sexual Function and distress by the Personal Distress Scale. Safety was assessed by adverse events, laboratory parameters and hormone levels. The TTP group demonstrated significant improvements in SSE (p = 0.0089) as well as in sexual desire (p = 0.0007) and reduced personal distress (p = 0.0024) versus placebo at 6 months (intent-to-treat analysis, n = 247). The results were significant for all three endpoints in the subgroup (n = 199) not using hormone therapy. Similar numbers of women treated with placebo and TTP discontinued (n = 39, 27.5% vs. n = 26, 20%), reported adverse events (including application site reactions) (n = 101, 71.1% vs. n = 81, 62.3%) and withdrew due to adverse events (n = 20, 14.1% vs. n = 9, 6.9%). No clinically relevant changes were noted in laboratory parameters. Serum free and total testosterone levels increased from baseline in the TTP group (geometric means 5.65 pg/ml and 67.8 ng/dl, respectively, at week 24) within the physiological range; no changes were seen in estradiol and sex hormone binding globulin levels. TTP was effective in treating HSDD and improving sexual function in this study of naturally menopausal women with and without concurrent hormone therapy.


Monitoring membranes: The exploration of biological bilayers with second harmonic generation.
Nature's seemingly controlled chaos in heterogeneous two-dimensional cell membranes stands in stark contrast to the precise, often homogeneous, environment in an experimentalist's flask or carefully designed material system. Yet cell membranes can play a direct role, or serve as inspiration, in all fields of biology, chemistry, physics, and engineering. Our understanding of these ubiquitous structures continues to evolve despite over a century of study largely driven by the application of new technologies. Here, we review the insight afforded by second harmonic generation (SHG), a nonlinear optical technique. From potential measurements to adsorption and diffusion on both model and living systems, SHG complements existing techniques while presenting a large exploratory space for new discoveries.


A New Statistics-Based Online Baseline Restorer for a High Count-Rate Fully Digital System.
The goal of this work is to develop a novel, accurate, real-time digital baseline restorer using online statistical processing for a high count-rate digital system such as positron emission tomography (PET). In high count-rate nuclear instrumentation applications, analog signals are DC-coupled for better performance. However, the detectors, pre-amplifiers and other front-end electronics would cause a signal baseline drift in a DC-coupling system, which will degrade the performance of energy resolution and positioning accuracy. Event pileups normally exist in a high-count rate system and the baseline drift will create errors in the event pileup-correction. Hence, a baseline restorer (BLR) is required in a high count-rate system to remove the DC drift ahead of the pileup correction. Many methods have been reported for BLR from classic analog methods to digital filter solutions. However a single channel BLR with analog method can only work under 500 kcps count-rate, and normally an analog front-end application-specific integrated circuits (ASIC) is required for the application involved hundreds BLR such as a PET camera. We have developed a simple statistics-based online baseline restorer (SOBLR) for a high count-rate fully digital system. In this method, we acquire additional samples, excluding the real gamma pulses, from the existing free-running ADC in the digital system, and perform online statistical processing to generate a baseline value. This baseline value will be subtracted from the digitized waveform to retrieve its original pulse with zero-baseline drift. This method can self-track the baseline without a micro-controller involved. The circuit consists of two digital counter/timers, one comparator, one register and one subtraction unit. Simulation shows a single channel works at 30 Mcps count-rate with pileup condition. 336 baseline restorer circuits have been implemented into 12 field-programmable-gate-arrays (FPGA) for our new fully digital PET system.


Elevated matrix metalloproteinase-9 and degradation of perineuronal nets in cerebrocortical multiple sclerosis plaques.
Matrix metalloproteinases (MMPs) degrade extracellular matrix; MMP activity, particularly of MMP-9, is elevated in the white matter in multiple sclerosis (MS) patients. The cerebral cortical extracellular matrix includes perineuronal nets (PNs) that surround parvalbumin-positive neurons (PV-positive neurons) and are important for their function. We measured active and total MMP-9 levels in postmortem homogenates of demyelinated and nondemyelinated cerebral cortical regions from 9MS and 7 control cases and assessed Wisteria floribunda agglutin (WFA)-positive PNs in paraffin sections from 15 MS and 6 controls and PV-positive neurons in sections from 26 MS and 6 controls. Active MMP-9 levels were higher in demyelinated than in nondemyelinated or control cortex (p < 0.05). The area fraction positive for WFA was lower in demyelinated than nondemyelinated MS or control cortex; the latter difference was significant (p < 0.05). Most PV-positive neurons in demyelinated but not intact cortex lackeda PN, and some showed perikaryal phosphorylated neurofilament protein accumulation. Loss of WFA-labeled PNs was not associated with reduced PV-positive neurons numbers. Thus, elevated MMP-9 in cortical plaques is associated with loss of PNs; PV-positive neurons are preserved but show abnormal neurofilament accumulations. Matrix metalloproteinase-mediated degradation of PNs in cortical plaques may, therefore, contribute to neuronal dysfunction and degeneration in MS patients.


Porphobilinogen-accumulating mutants of Salmonella typhimurium LT2.
Four independent porphobilinogen-accumulating mutants of Salmonella typhimurium LT2 were isolated by selecting for dwarf colony formation on neomycin agar media. Cell-free extracts of the parent strain, but not of the mutants, were able to convert 5-aminolaevulinic acid or porphobilinogen to porphyrins. The results indicated that the mutants were deficient in uroporphyrinogen I synthase (EC. 4.3.I. 8) activity: these are the first mutants of this type reported in S. typhimurium LT2. Mapping of the hemC locus (for uroporphyrinogen I synthase) by F-mediated conjugation and by P22-mediated transduction showed the gene sequence ilvEDAC-hemC-cya-metE.


Finding Aquaporins in Annelids: An Evolutionary Analysis and a Case Study.
Aquaporins (AQPs) are a family of membrane channels facilitating diffusion of water and small solutes into and out of cells. Despite their biological relevance in osmoregulation and ubiquitous distribution throughout metazoans, the presence of AQPs in annelids has been poorly investigated. Here, we searched and annotated Aqp sequences in public genomes and transcriptomes of annelids, inferred their evolutionary relationships through phylogenetic analyses and discussed their putative physiological relevance. We identified a total of 401 Aqp sequences in 27 annelid species, including 367 sequences previously unrecognized as Aqps. Similar to vertebrates, phylogenetic tree reconstructions clustered these annelid Aqps in four clades: AQP1-like, AQP3-like, AQP8-like and AQP11-like. We found no clear indication of the existence of paralogs exclusive to annelids; however, several gene duplications seem to have occurred in the ancestors of some Sedentaria annelid families, mainly in the AQP1-like clade. Three of the six Aqps annotated in Alitta succinea, an estuarine annelid showing high salinity tolerance, were validated by RT-PCR sequencing, and their similarity to human AQPs was investigated at the level of "key" conserved residues and predicted three-dimensional structure. Our results suggest a diversification of the structures and functions of AQPs in Annelida comparable to that observed in other taxa.


Pharmacotherapy quality improvement in the managed care setting.
Physician partnering, value-added relationships, and integrated delivery systems with risk/incentive arrangements are emerging in managed care. These arrangements require time-sensitive program information that can be used in continuous quality improvement programs. Components of any clinical intervention program designed to improve the quality of pharmacotherapy are on a continuum, from those that are primarily individual patient focused to those that are disease or system/process focused. Components include case management, patient profile review, brand to generic drug conversion, targeted therapeutic evaluations, provider profiling, and pharmacoeconomic analysis. All health care professionals involved in pharmacotherapy need to be involved in implementation of these quality improvement programs.


Local reports of climate change impacts in Sierra Nevada, Spain: sociodemographic and geographical patterns.
While we know that climate change is having different impacts on various ecosystems and regions of the world, we know less how the perception of such impacts varies within a population. In this study, we examine patterns of individual variation in climate change impacts reports using data from a sample (n = 238) drawn from 33 mountainous municipalities of Sierra Nevada, Spain. Sierra Nevada inhabitants report multiple climate change impacts, being the most frequently reported changes in snowfall and snow cover, abundance of terrestrial fauna, freshwater availability, and extreme temperatures. Reports of climate change impacts vary according to informants' sociodemographic characteristics and geographical location. People with life-long bonds with the environment and higher connection and dependence upon ecosystem services report more climate change impacts than other informants, as do people with lower level of schooling. We also found that reports of climate change impacts vary according to geographic areas, which reinforces the idea that climate change generates differentiated impacts even at small geographical scales. Understanding intracultural variation in reports of climate change impacts not only gives an enriched picture of the human dimensions of climate change but might also help design more targeted mitigation and adaptation responses. The online version contains supplementary material available at 10.1007/s10113-022-01981-5.


Health-related quality of life and prevalence of six chronic diseases in homeless and housed people: a cross-sectional study in London and Birmingham, England.
To compare health-related quality of life and prevalence of chronic diseases in housed and homeless populations. Cross-sectional survey with an age-matched and sex-matched housed comparison group. Hostels, day centres and soup runs in London and Birmingham, England. Homeless participants were either sleeping rough or living in hostels and had a history of sleeping rough. The comparison group was drawn from the Health Survey for England. The study included 1336 homeless and 13 360 housed participants. Chronic diseases were self-reported asthma, chronic obstructive pulmonary disease (COPD), epilepsy, heart problems, stroke and diabetes. Health-related quality of life was measured using EQ-5D-3L. Housed participants in more deprived neighbourhoods were more likely to report disease. Homeless participants were substantially more likely than housed participants in the most deprived quintile to report all diseases except diabetes (which had similar prevalence in homeless participants and the most deprived housed group). For example, the prevalence of chronic obstructive pulmonary disease was 1.1% (95% CI 0.7% to 1.6%) in the least deprived housed quintile; 2.0% (95% CI 1.5% to 2.6%) in the most deprived housed quintile; and 14.0% (95% CI 12.2% to 16.0%) in the homeless group. Social gradients were also seen for problems in each EQ-5D-3L domain in the housed population, but homeless participants had similar likelihood of reporting problems as the most deprived housed group. The exception was problems related to anxiety, which were substantially more common in homeless people than any of the housed groups. While differences in health between housed socioeconomic groups can be described as a 'slope', differences in health between housed and homeless people are better understood as a 'cliff'.


Development of a C-reactive protein quantification method based on flow rate measurement of an ink solution pushed out by oxygen gas generated by catalase reaction.
A novel determination method for protein biomarkers based on on-chip flow rate measurement was developed using a microchip with organic photodiodes (OPDs). This quantitative method is based on the flow rate measurement of an ink solution pushed out by oxygen gas generated through catalase reaction. The amount of oxygen gas generated in the sample reservoir is dependent on the concentration of the analyte; therefore, the flow rate of the ink solution is also dependent on the concentration of the analyte. The concentration of the analyte can thus be estimated by measurement of the ink solution flow rate. The ink solution flow rate was estimated by measuring the migration time of the ink solution between two points using two OPDs placed below the microchannel. The principle of this method was demonstrated by the measurement of catalase using the microchip. In addition, the developed method was applied to the determination of C-reactive protein (CRP), a biomarker of inflammation, based on a catalase-linked immunosorbent assay (C-LISA). The limit of detection for CRP was 0.20 µg/mL. The method was also applied to the determination of CRP in human serum, and the quantitative values obtained by this method were in excellent agreement with those obtained by the conventional enzyme-linked immunosorbent assay (ELISA) method. The developed method does not require a photodetector with high sensitivity and is thus capable of downsizing; therefore, this will be useful for on-site analyses such as point-of-care testing and field measurements.


Keto fatty acids not containing doubly allylic methylenes are lipoxygenase substrates.
The soybean lipoxygenase I oxygenates the unusual substrate 12-keto-(9Z)-octadecenoic acid methyl ester as indicated by oxygen uptake and spectral changes of the incubation mixture. The main oxygenation products have been isolated by HPLC and identified as 9,12-diketo-(10E)-octadecenoic acid methyl ester and 12-keto-(10E)-dodecenoic acid methyl ester by UV and IR spectroscopy, cochromatography with an authentic standard, gas chromatography/mass spectroscopy, and 1H NMR. In the formation of both compounds the oxygenase and hydroperoxidase activities of the enzyme appear to be involved. These data and the earlier results on the oxygenation of furanoic fatty acids (Boyer et al., 1979) indicate that the lipoxygenase reaction is not restricted to substrates containing a 1,4-pentadiene structure.


Rapid dry plasma thawing system: An alternative to conventional thawing baths.
Fresh frozen plasma (FFP) should be thawed before transfusing to the patient. Prolonged or uncontrolled thawing can denature plasma proteins. The potential risk of contamination by wet thawing had always been a point of concern. Here, we compared and evaluated the effect of thawing on clotting factor activities by two different methods (wet and dry) and other factors such as risk of bacterial contamination, throughput, turnaround time, and efficacy of thawing. All FFPs were prepared from Group O donors and stored at -40°C. Twenty-one FFPs were thawed in Plasmatherm II at 45°C for 15 min and another 21 were thawed in thawing bath at 37°C for 20-30 min randomly. Analysis of prothrombin time, activated partial thromboplastin time, fibrinogen, and factor VIII was done in ACL TOP 300 (IL) at the time of preparation and immediately after thawing of FFPs. Volume, duration of thawing, ease of use, accessibility, and equipment maintenance were also compared. There was a statistically significant difference in coagulation parameters after thawing in both methods compared to the time of preparation (P < 0.05), but all values were within normal limits. There was no significant difference in coagulation parameters between the two methods (P > 0.05). Mixed bacterial growth was observed from swabs taken from the water bath. Plasmatherm II can be a good alternative to water bath to rapidly thaw FFPs by preserving coagulation factors and eliminating the risk of bacterial contamination.


A Public health perspective on mitigating the global burden of chronic liver disease.
Chronic liver disease (CLD) is a significant global health problem. Epidemiological trends do not show improvement in CLD incidence but rather a shift in etiologies, with steatotic liver disease (SLD) from metabolic dysfunction and alcohol becoming increasingly important causes. Consequently, there is a pressing need to develop a comprehensive public health approach for SLD. To that end, we propose a public health framework for preventing and controlling SLD. The framework is anchored on evidence linking physical inactivity, unhealthy dietary patterns, alcohol use, and obesity with both incidence and progression of SLD. Guided by the framework, we review examples of federal/state, community, and individual-level interventions with the potential to address these determinants of SLD. Ultimately, mitigating SLD's burden requires primary risk factor reduction at multiple socioecological levels, by scaling up the World Health Organization's "best buys", in addition to developing and implementing SLD specific control interventions.


Detecting causality from online psychiatric texts using inter-sentential language patterns.
Online psychiatric texts are natural language texts expressing depressive problems, published by Internet users via community-based web services such as web forums, message boards and blogs. Understanding the cause-effect relations embedded in these psychiatric texts can provide insight into the authors' problems, thus increasing the effectiveness of online psychiatric services. Previous studies have proposed the use of word pairs extracted from a set of sentence pairs to identify cause-effect relations between sentences. A word pair is made up of two words, with one coming from the cause text span and the other from the effect text span. Analysis of the relationship between these words can be used to capture individual word associations between cause and effect sentences. For instance, (broke up, life) and (boyfriend, meaningless) are two word pairs extracted from the sentence pair: "I broke up with my boyfriend. Life is now meaningless to me". The major limitation of word pairs is that individual words in sentences usually cannot reflect the exact meaning of the cause and effect events, and thus may produce semantically incomplete word pairs, as the previous examples show. Therefore, this study proposes the use of inter-sentential language patterns such as ≪broke up, boyfriend>, <life, meaningless≫ to detect causality between sentences. The inter-sentential language patterns can capture associations among multiple words within and between sentences, thus can provide more precise information than word pairs. To acquire inter-sentential language patterns, we develop a text mining framework by extending the classical association rule mining algorithm such that it can discover frequently co-occurring patterns across the sentence boundary. Performance was evaluated on a corpus of texts collected from PsychPark (http://www.psychpark.org), a virtual psychiatric clinic maintained by a group of volunteer professionals from the Taiwan Association of Mental Health Informatics. Experimental results show that the use of inter-sentential language patterns outperformed the use of word pairs proposed in previous studies. This study demonstrates the acquisition of inter-sentential language patterns for causality detection from online psychiatric texts. Such semantically more complete and precise features can improve causality detection performance.


Expression of monomorphic arylamine N-acetyltransferase (NAT1) in human leukocytes.
The expression of arylamine N-acetyltransferase (NAT) in leukocytes was investigated using p-aminobenzoic acid (PABA) and sulfamethazine (SMZ), substrates which are preferentially acetylated by the monomorphic NAT1 and polymorphic NAT2 enzymes, respectively. Activity towards both substrates was detected in mononuclear leukocytes (MNL; preparation containing approximately 80% lymphocytes), monocytes and neutrophils. PABA and SMZ acetylation rates were highly correlated in each of the isolated cell types. The NAT in leukocytes displayed a much higher affinity and turnover rate for the acetylation of PABA than for SMZ. These kinetic characteristics suggest that the acetylating activity in human leukocytes is predominantly attributable to the monomorphic enzyme NAT1. Neutrophils showed evidence of biphasic kinetics for SMZ which would indicate the coexpression of NAT1 and low levels of the polymorphic enzyme, NAT2. NAT activity in MNL was not influenced by the acetylator phenotype of the individual. There was, however, a significant correlation between NAT activity in MNL and the in vivo acetylation (urinary metabolite ratio) of p-aminosalicylic acid, which is monomorphically acetylated in humans. The expression of NAT1 in leukocytes and the virtuall absence of NAT2 may have important toxicological implications. The in vitro/in vivo correlation suggests that leukocytes may be a useful marker of systemic NAT1 activity.


Utilization of a hybrid sequencing batch reactor (HSBR) as a decentralized system of domestic wastewater treatment.
This paper presents the experiments carried out in a hybrid sequencing batch reactor (HSBR), used for biological treatment of sewage. The HSBR was built in a cylindrical shape and made of stainless steel, with a volume of 1.42 m3. Besides the biomass in suspension, the reactor also carried fixed biomass (hybrid process), adhered in the support material. This consisted of a nylon net disposed in a grille for biofilm biomass adhesion. The reactor worked fully automated in operational cycles of maximum 8 hours each, presenting the following phases: filling, anoxic, aerobic, settle and draw of treated effluent, with 3 fillings per cycle. Increasing organic loads (0.14 to 0.51 kg TCOD/m3 day) and ammonium loads (0.002 to 0.006 kg NH4-N/m3.day) were tested. We monitored the reactor's performance by measuring the liquid phase (COD, pH, temperature, DO, nitrogen and phosphorus) during the cycles and by measuring the sludge through respirometric tests. The results obtained demonstrated TCOD removal efficiency between 73 and 96%, and ammonium removal efficiency between 50 and 99%. At the end of the cycles, the effluent presented ammonium concentration <20 mg/L, meeting the Brazilian environmental legislation standards (CONAMA 357/2005) regarding discharges into the water bodies. Respirometric tests showed biomass dependency on FCOD concentrations. Results have demonstrated the potential of this type of reactor for decentralized treatment of domestic wastewater.


Sarcoplasmic-reticulum biogenesis in contraction-inhibited skeletal-muscle cultures.
We have previously shown that inhibition of the spontaneous contractile activity of cultured embryonic-chick skeletal-muscle fibres with tetrodotoxin (TTX) leads to decreased sarcoplasmic-reticulum Ca(2+)-transport rates and steady-state concentrations of the high-energy Ca(2+)-ATPase phosphoenzyme intermediate [Charuk & Holland (1983) Exp. Cell Res. 144, 143-157]. In the present study we used a monoclonal antibody to the Ca(2+)-ATPase to show that there is a decreased amount of enzyme accumulated by contraction-inhibited myotubes. Indirect immunofluorescence microscopy using the monoclonal antibody to the Ca(2+)-ATPase also revealed a disordered subcellular organization of the sarcotubular system in contraction-inhibited myotubes. The biogenesis of sarcoplasmic-reticulum proteins in TTX-paralysed myofibres was studied by labelling cells with [35S]methionine before isolation of the active Ca(2+)-pump membrane fraction. Protein turnover was selectively increased in that fraction from TTX-treated muscle cultures. Electrophoretic analysis and quantitative fluorography confirmed that decreased accumulation of the Ca(2+)-ATPase enzyme in contraction-inhibited myotubes was associated with increased turnover of this protein. The present results demonstrate that biogenesis of the sarcoplasmic-reticulum Ca(2+)-ATPase is regulated by the contractile activity of skeletal-muscle fibres.


A fiber modified adenovirus vector that targets to the EphrinA2 receptor reveals enhanced gene transfer to ex vivo pancreatic cancer.
Pancreatic cancer is an aggressive malignancy with a dismal prognosis. To improve treatment options new treatments, such as adenoviral (Ad) gene therapy are necessary. However, low expression of the coxsackie and adenovirus receptor (CAR) in pancreatic cancer cells (PC) limits the therapeutic efficacy of these vectors. The aim of this study was to improve transduction of PC by recombinant adenoviruses by inserting peptides into the HI loop that binds to receptors highly expressed on pancreatic cancer and were shown to target these carcinomas in vivo. We report the successful incorporation into the HI loop of peptide Tyr-Ser-Ala (YSA), a peptide ligand targeting the EphrinA2 (EphA2) receptor, and K237, a peptide targeting to the vascular endothelial growth factor receptor-II (VEGFRII). Subsequently, we showed that both peptides enhanced the transduction of a number of human PC lines that abundantly express the targeted receptor. Additional competition studies confirmed that the YSA peptide redirects Ad-YSA from CAR and specifically targets the EphA2 receptor. Due to this transduction efficiency of Ad-YSA is increased not only in human pancreatic cancer cell lines but more importantly also in pancreatic cancer resection specimens. Since the YSA peptide has been shown to specifically target pancreatic cancer in patients, it may be expected that Ad-YSA will also display increased tropism for this tumour.


Formulation Development and in vitro Characterization of Proliposomes for Topical Delivery of Aceclofenac.
Non-steroidal antiinflammatory drugs are routinely prescribed for the patients with rheumatic disease and such patients are at increased risk of serious gastrointestinal complications, when non-steroidal antiinflammatory drugs administered by oral route. The aim of the present study was to develop and characterized a vesicular drug carrier system (proliposome) for topical delivery of aceclofenac to overcome the problems related with oral route. Aceclofenac proliposome were prepared by the film-deposition on carriers method and characterized for size and surface morphology, drug content in both proliposomes and liposomal system, percent yield, in vitro drug release studies and drug permeation studies. The prepared system was also characterized for drug-excipients interaction by Fourier transform infrared spectrophotometer and stability studies. The size and surface morphology were studied using optical microscopy, scanning electron microscopy and transmission electron microscopy. A spherical shape of reconstituted aceclofenac liposome with an average vesicular size of about 500 nm was observed in photomicrographs. The maximum entrapment efficiency of reconstituted liposomes was 80.31% whereas the drug content in proliposomes was found to be more than 90%. In vitro release of drug was significantly retarded indicating sustained release of aceclofenac from proliposomes. Stability study was performed at various temperatures indicating that aceclofenac proliposomes are stable at lower temperature.


Disclosing odd symmetry, strain driven magnetic response of Co on Pt/PMN-PT (0 1 1).
An odd-symmetry magnetic response of multiferroic composites comprising ultrathin Co layers on Pt electrodes on [Pb(Mg0.33Nb0.67)O3](1-x)[PbTiO3] x (PMN-PT) (0 1 1) piezoelectric substrates is observed upon electrical poling of the PMN-PT substrates: the magnetic easy axis of the Co rotates by 90° in-plane between oppositely poled ferroelectric states, mimicking the signature of a surface polarization charge driven effect, which however can be excluded from the presence of the thick Pt interlayer. The origin for this unexpected behavior is as an odd symmetry piezoelectric response of the PMN-PT substrate, as indicated by x-ray diffraction with applied poling, in combination with conventional magnetoelastic coupling. Ferroelectric characterization reveals corresponding features, possibly related to an unswitchable polarization component.


Analysis of the CO dehydrogenase/acetyl-coenzyme A synthase operon of Methanosarcina thermophila.
The cdhABC genes encoding the respective alpha, epsilon, and beta subunits of the five-subunit (alpha, beta, gamma, delta, and epsilon) CO dehydrogenase/acetyl-coenzyme synthase (CODH/ACS) complex from Methanosarcina thermophila were cloned and sequenced. Northern (RNA) blot analyses indicated that the cdh genes encoding the five subunits and an open reading frame (ORF1) with unknown function are cotranscribed during growth on acetate. Northern blot and primer extension analyses suggested that mRNA processing and multiple promoters may be involved in cdh transcript synthesis. The putative CdhA (alpha subunit) and CdhB (epsilon subunit) proteins each have 40% identity to CdhA and CdhB of the CODH/ACS complex from Methanosaeta soehngenii. The cdhC gene encodes the beta subunit (CdhC) of the CODH/ACS complex from M. thermophila. The N-terminal 397 amino acids of CdhC are 42% identical to the C-terminal half of the alpha subunit of CODH/ACS from the acetogenic anaerobe Clostridium thermoaceticum. Sequence analysis suggested potential structures and functions for the previously uncharacterized beta subunit from M. thermophila. The deduced protein sequence of ORF1, located between the cdhC and cdhD genes, has 29% identity to NifH2 from Methanobacterium ivanovii.


Distribution of hepatitis C virus genotypes determined by line probe assay in patients with chronic hepatitis C seen at tertiary referral centers in the United States. Hepatitis Interventional Therapy Group.
To 1) verify the validity of a new line probe assay for hepatitis C virus (HCV) genotyping and 2) determine the distribution of HCV genotypes and the association between HCV genotype and clinical variables in patients with chronic hepatitis C seen in tertiary referral centers in the United States. Retrospective cross-sectional analysis. 438 patients with chronic hepatitis C from 10 tertiary referral centers. The validity of the line probe assay was first verified against a panel of serum specimens that had previously been characterized by six different HCV genotyping methods. Specimens from all 438 patients were then genotyped using this line probe assay. The associations between HCV genotype and clinical variables were examined using analysis of variance. Pairwise testing was used when the F test showed a statistically significant difference. Nonparametric alternatives were used for variables for which normality could not be assumed. The line probe assay was quick and reproducible, and it showed good concordance with other tests. In our sample, the proportions of patients with HCV types 1, 2, 3, and 4 were 71.5%, 13.5%, 5.5%, and 1.1%, respectively. Subtypes 1a and 1b were seen in approximately equal proportions of patients with HCV type 1. Mixed infection was detected in 3.7% of specimens, and 4.8% of specimens either had negative results on polymerase chain reaction or could not be typed. A higher proportion of patients with HCV type 1 than of patients with HCV-type 1 had acquired HCV through transfusion of blood products (50% compared with 25%; P < 0.001). Patients with HCV type 1 also had a longer estimated duration of infection compared with patients with HCV type 3 (P = 0.004) and type 4 (P = 0.049). Disease activity did not differ among patients infected with HCV types 1, 2, or 3. Levels of viremia were similar in patients with HCV types 1, 2, or 3, but patients with HCV type 4 had a lower level of viremia than did patients with HCV type 1 (P = 0.047). The line probe assay can be used in patients with chronic HCV infection in the United States. In patients with chronic hepatitis C referred to tertiary centers in the United States, type 1 is the most common HCV genotype. Disease activity and viremia levels do not differ among patients chronically infected with HCV types 1, 2, or 3.


The NEDD8-activating enzyme inhibitor pevonedistat activates the eIF2α and mTOR pathways inducing UPR-mediated cell death in acute lymphoblastic leukemia.
Acute lymphoblastic leukemia (ALL) is the leading cause of cancer-related death in children, and cure rates for adults remain dismal. Further, effective treatment strategies for relapsed/refractory ALL remain elusive. We previously uncovered that ALL cells are prone to apoptosis via endoplasmic reticulum (ER) stress/unfolded protein response (UPR)-mediated mechanisms. We investigated the antineoplastic activity of pevonedistat®, a novel NEDD8-activating enzyme inhibitor that targets E3 cullin-RING ligases (CRLs) dependent proteasomal protein degradation, in ALL. Herein, we report that pevonedistat induces apoptosis in ALL cells by dysregulating the translational machinery leading to induction of proteotoxic/ER stress and UPR-mediated cell death. Mechanistically, pevonedistat led to P-eIF2a dephosphorylation causing atypical proteotoxic/ER stress from failure to halt protein translation via the UPR and upregulation of mTOR/p70S6K. Additional studies revealed that pevonedistat re-balanced the homeostasis of pro- and anti-apoptotic proteins to favor cell death through altered expression and/or activity of Mcl-1, NOXA, and BIM, suggesting that pevonedistat has a "priming" effect on ALL by altering the apoptotic threshold through modulation of Mcl-1 activity. Further, we demonstrated that pevonedistat synergizes with selected anti-leukemic agents in vitro, and prolongs survival of NSG mice engrafted with ALL cells, lending support for the use of pevonedistat as part of a multi-agent approach.


An European inter-laboratory validation of alternative endpoints of the murine local lymph node assay: first round.
The new OECD guideline 429 (skin sensitization: local lymph node assay) is based upon a protocol, which utilises the incorporation of radioactivity into DNA as a measure for cell proliferation in vivo. The guideline also enables the use of alternative endpoints in order to assess draining lymph node (LN) cell proliferation. Here we describe the first round of an inter-laboratory validation of alternative endpoints in the LLNA conducted in seven laboratories. The validation study was managed and supervised by the Swiss Agency for Therapeutic Products, Swissmedic. Statistical analyses of all data were performed by an independent centre at the University of Bern, Department of Statistics. Ear-draining, LN weight and cell count were used to assess proliferation instead of radioactive labeling of lymph node cells. In addition, the acute inflammatory skin reaction was measured by ear swelling and weight of circular biopsies of the ears to identify skin irritating properties of the test items. Hexylcinnamaldehyde (HCA) and three blinded test items were applied to female, 8--10 weeks old NMRI and BALB/c mice. Results were sent via the independent study coordinator to the statistician. The results of this first round showed that the alternative endpoints of the LLNA are sensitive and robust parameters. The use of ear weights added an important parameter assessing the skin irritation potential, which supports the differentiation of pure irritative from contact allergenic potential. There were absolute no discrepancies between the categorisation of the three test substances A--C determined by each single participating laboratories. The results highlighted also that many parameters do have an impact on the strength of the responses. Therefore, such parameters have to be taken into consideration for the categorisation of compounds due to their relative sensitizing potencies.


Process evaluation results from a school- and community-linked intervention: the Trial of Activity for Adolescent Girls (TAAG).
Process evaluation is a component of intervention research that evaluates whether interventions are delivered and received as intended. Here, we describe the process evaluation results for the Trial of Activity for Adolescent Girls (TAAG) intervention. The intervention consisted of four synergistic components designed to provide supportive school- and community-linked environments to prevent the decline in physical activity in adolescent girls. Process evaluation results indicate that the intervention components were delivered from intervention staff to teachers with high fidelity (84-97%) to the protocol and with lower fidelity (range: 18-93%) from teachers to students. Physical activity programs for girls, a unique feature of the TAAG intervention, increased from a mean of 10 programs per school to a mean of 16 and 15 in years 1 and 2, respectively, in intervention schools, with no change in control schools. These findings suggest that a multicomponent school- and community-based physical activity intervention can be delivered with fidelity and result in a middle school environment that supports physical activity for girls.


Liposomes as carriers of antitumor agents: toward a clinical reality.
For several years, liposomes have been explored as carriers of antitumor agents. Liposomes can be administered intravenously to carry lipid-soluble drugs, to improve drug stability, to target organs with fenestrated capillaries, to achieve an intravascular slow drug-release system, and to reduce drug levels in certain organs sensitive to toxicity. They also have significant potential for local therapy when administered subcutaneously or intraperitoneally. The preclinical development of liposome-entrapped antitumor agents has been, in general, slowed by cumbersome formulation problems. However, during the last few years, new drug loading techniques and a better selection of the drugs for liposome entrapment have resulted in pharmaceutically acceptable liposomal formulations or antitumor agents. Three preparations have been approved by the Federal Drug Administration, two containing doxorubicin, and the other a new lipophilic cisplatin analogue [cis-bis-neodecanoato-trans-R,R-1,2-diamino-cyclohexane platinum (II)]. In preclinical studies, these preparations were shown to be less toxic than the parent compounds and more active in models of liver micrometastases. Clinical Phase I and II studies with these formulations are now in progress. Although liposomal antitumor agents have no established role in the anticancer armamentarium at this stage, the information available suggests that they may improve the therapeutic index or broaden the applications of available antitumor agents and possibly act as carriers for newly designed liposome-dependent antitumor agents.


Amyloid oligomers exacerbate tau pathology in a mouse model of tauopathy.
We aimed to investigate the influence of oligomeric forms of β-amyloid (Aβ) and the influence of the duration of exposure on the development of tau phosphorylation. Aβ oligomers were injected intracranially either acutely into 5-month-old rTg4510 mice and tissue was collected 3 days later, or chronically into 3-month-old mice and tissue was collected 2 months later. Several forms of phosphorylated tau (p-tau), GSK3 (glycogen synthase kinase-3) and microglial and astrocyte activation were measured. Acute injections of Aβ oligomers had no effect on p-tau epitopes but did result in elevation of phosphorylated/activated GSK3 (pGSK3). Chronic infusion of Aβ oligomers into the right hippocampus resulted in 3- to 4-fold elevations in several p-tau isoforms with no changes in total tau levels. A significant elevation in pGSK3 accompanied these changes. Microglial staining with CD68 paralleled the increase in tau phosphorylation, however, CD45 staining was unaffected by Aβ. Control experiments revealed that the infusion of Aβ from the minipumps was largely complete by 10 days after implantation. Thus, the elevation in p-tau 2 months after implantation implies that the changes are quite persistent. Soluble Aβ(1-42) oligomers have long-lasting effects on tau phosphorylation in the rTg4510 model, possibly due to elevations in GSK3. These data suggest that even brief elevations in Aβ production, may have enduring impact on the risk for tauopathy.


Retention of HIV exposed infants in care at Arua regional referral hospital, Uganda: a retrospective cohort study.
Retention of HIV Exposed Infants (HEIs) in care ensures adequate care. Data on retention of HEIs at large referral hospitals in Uganda is limited. We investigated the retention level of HEIs and associated factors. We conducted a retrospective cohort study on 352 HEIs in care (January 2014 and April 2015) at Arua Regional Referral Hospital, North-western Uganda. Electronic medical data were retrieved and analyzed with Stata. Chi-square, Fisher's exact, and Students t-tests were used for bivariate analysis. Logistic regression was performed to determine factors independently associated with retention. 236 (67.0%) HEIs were delivered in a health facility and 306 (86.9%) received Nevirapine prophylaxis from birth until 6-weeks. Of mothers, 270 (76.7%) were 25-46 years, 202 (57.4%) attended antenatal care (ANC) at recent pregnancy, and 328 (93.2%) were on life-long anti-retroviral therapy. At 18-months, 277 (78.7%) HEIs were retained in care. Maternal age (25-46 years) (Adjusted Odds Ratio (AOR), 2.32; 95% CI, 1.32-4.06), ANC attendance during recent pregnancy (AOR, 2.01; 95% CI, 1.19-4.3.41) and Nevirapine prophylaxis initiation from birth until 6-weeks (AOR, 3.07; 95% CI, 1.50-6.26) were associated with retention. Retention was suboptimal. Older maternal age, ANC visits at last pregnancy, and timely NVP initiation increased retention.


Potential immunological advantage of intravenous mycophenolate mofetil with tacrolimus and steroids in primary deceased donor liver transplantation and live donor liver transplantation without antibody induction.
With the current immunosuppressive regimens, graft loss secondary to immunological reasons after successful liver transplantation is a rarity; acute rejections, however, do occur, with the majority of them being steroid-responsive. The aim of the present study is to examine the rate of acute rejection with tacrolimus, intravenous (IV) mycophenolate mofetil (MMF), and steroids in primary deceased donor liver transplant (DDLT) and live donor liver transplant (LDLT) recipients. During the year 2005, 130 patients (mean age: 54.9 +/- 10.8, males: 84, females: 46, 112 DDLT and 18 LDLT) received primary liver transplantation. They were followed up for the incidence of acute rejection in the first 12 months. Liver biopsies were performed as clinically indicated; protocol liver biopsies were never performed. A total of 127 liver biopsies were performed. Thirty-two had a rejection activity index (RAI) score of > or =3, of which 24 biopsies in 20 patients were not treated with a steroid bolus. Eight (6.1%) patients (mean RAI score: 5.1 +/- 1.4) received 750 to 1500 mg of methylprednisolone over 3 days. Out of these, 2 were noncompliant, 4 were off MMF, and 1 was on cyclosporine. All patients responded to steroid therapy. None of the patients required any antibody preparation. In conclusion, IV MMF with tacrolimus and steroids is useful and required antirejection therapy in 6.1% of liver transplant recipients.


Determination of the sizes of the opioid receptors in their membrane environment by radiation inactivation.
Opioids, like other drugs, are thought to initiate their effects by association with their specific receptors. However, very little is known about the opioid receptor as a molecular entity. The binding components have been solubilized in detergent and purified by different approaches, but the molecular size of soluble opioid receptor complexes reported by different groups varied from 23,000 to 750,000. In this study, the technique of radiation inactivation by gamma rays was used to investigate the apparent size of the opioid receptor in rat brain membranes under different conditions. The molecular sizes of opioid receptor complexes were estimated as 313,000 +/- 13,500 in the presence of [D-Ala2, D-Leu5] enkephalin, NaCl and Gpp (NH)p; as 165,000 +/- 8,500 in the presence of NaCl only, or of both NaCl and Gpp (NH)p; as 217,000 +/- 6,600 in the presence of Gpp (NH)p only; and as 286,000 +/- 60,900 in the presence of MgCl2 only. A simple model has been proposed to explain these different apparent target sizes of opioid receptors obtained under different conditions.


Lesions of A1 noradrenergic cells affect AVP release and heart rate during hemorrhage.
The role of A1 noradrenergic cells of the ventrolateral medulla in the changes in mean arterial pressure (MAP), heart rate (HR), and plasma arginine vasopressin (AVP) after slow continuous hemorrhage (2% blood vol/min up to 35%) was examined by comparing responses in conscious rabbits before and 3 wk after a sham operation or A1 lesions. In the control experiments, MAP fell minimally up to the withdrawal of 20% of blood volume after which it fell abruptly to 20-30 mmHg below control by the 35% level. Plasma AVP increased nonlinearly during progressive hemorrhage with significant increases occurring only after 25% of blood volume was removed. In contrast, HR increased linearly after the onset of bleeding. After A1 lesions, which destroyed 84% (range 80-94%) of the noradrenergic cells, the amount of AVP released and the tachycardia during hemorrhage were reduced by 83 and 61%, respectively (P less than 0.005), but the fall in MAP was minimally affected. Basal values of MAP, HR, or plasma AVP were not affected by the lesions. These results suggest that during hemorrhage in conscious rabbits A1 noradrenergic neurons are important for the secretion of AVP and the reflex tachycardia but play little role in the maintenance of blood pressure.


The effect of donor age on the proliferative response of human and mouse keratinocytes to phorbol, 12-myristate, 13-acetate.
Epidermal keratinocytes grow clonally when provided with a 3T3 feeder layer and medium supplemented with 20% foetal bovine serum, hydrocortisone and cholera toxin. In this culture system the proliferative response of freshly isolated human and mouse keratinocytes to a short exposure (24 h) of the tumour promoter phorbol, 12-myristate, 13-acetate (PMA) was dependent on the donor age but was independent of the species or the biopsy site. Human keratinocytes from early (16-18 week) foetal donors displayed a strong proliferative response to PMA as determined by a 5- to 7-fold increase in colony number and an increase in the average colony size. In contrast, adult and juvenile keratinocytes of all ages from both mice and humans displayed an inhibition of colony forming efficiency and a reduction in colony size. When continuously passaged in culture human foetal keratinocytes gradually changed the pattern of their response to PMA so that they were inhibited from growing rather than being stimulated and after 21 days (three passages) their response was quantitatively similar to juvenile keratinocytes. Co-culture of juvenile keratinocytes with irradiated foetal keratinocytes did not alter their response to PMA so that the observed proliferative response of foetal keratinocytes to PMA is not readily explained by the autosecretion of mitogens or other regulatory molecules by these cells. Late foetal (17 days gestation), neonatal and post-neonatal (5-10 days old) mouse keratinocytes were also inhibited from growing by PMA but the magnitude of the effect was directly related to the age of the mouse and was in all cases less than that observed with adult mice. The relationship of these results to the mechanism of action of phorbol esters in epidermis is discussed.


Effect of UV-light on the uniaxial tensile properties and structure of uncoated and TiO2 coated Bombyx mori silk fibers.
The effect of UV-light on the uniaxial tensile properties and the structure of uncoated and TiO2 coated silk fibers in the bave form by using sol-gel method was investigated with tensile testing and FT-IR/ATR spectroscopy methods after the silk filaments were exposed to UV-light with high intensity of 760W/m(2) for different times from 0.5h to 1day. It was clearly observed that TiO2 coating considerably increased the Young's modulus of the uncoated silk single filament by around 17% before the UV-irradiation. The yield point and the post yield region disappeared on the stress-strain curves of both uncoated and TiO2 coated silk filaments after UV-irradiation time higher than 1h. Except for the Young's modulus, most of the tensile characteristics of both uncoated and TiO2 coated silk filaments decreased remarkably with increasing UV-irradiation time, e.g., after 1h irradiation, although the Young's modulus slightly changed and ultimate tensile strength decreased by only around 18% and 23%, for the uncoated and TiO2 coated silk filaments, respectively; breaking extension decreased dramatically by 67% and 72%, respectively, for uncoated and TiO2 coated silk filaments. Only the Young's modulus of TiO2 coated silk filaments which can be considered as a more stable tensile characteristic became significantly higher than that of the uncoated silk filaments with increasing UV-irradiation time. After 1day irradiation, even though the uncoated silk filaments could not be tested and completely lost of their fiber properties, the TiO2 coated silk filaments showed a stress-strain curve in initial elastic region with Young's modulus of ∼13GPa which indicates considerable protective effect of TiO2 on the silk fiber structure, especially on the β-sheet microcrystals against UV-radiation. The FT-IR/ATR spectral results showed that significant photodegradation took place in not only crystalline but also amorphous regions which were deduced from the decrease in the absorbance ratios of the bands assigned to CH3 rocking, Cα-Cβ, Cα-C stretching vibrations in β-sheet crystalline regions as well as the Amide I, II, and III bands for both crystalline and amorphous regions. Even though the ratio of crystalline to amorphous regions in uncoated silk filaments decreased significantly, the ratio in TiO2 coated silk filaments became almost constant with increasing UV-irradiation time which may indicate more stable β-sheet microcrystals against photodegradation.


Identification of genetic heterogeneity in the Cryptosporidium parvum ribosomal repeat.
Oocysts of the protozoan parasite Cryptosporidium parvum are found in most surface waters and can contaminate municipal water supplies, as demonstrated by recent outbreaks of cryptosporidiosis. A method capable of fingerprinting C. parvum isolates from the environment would facilitate the study of epidemiology and transmission cycles and aid in the implementation of preventive measures to reduce water contamination by oocytes. In this study, we report polymorphism in C. parvum isolates on the basis of analysis of random amplified polymorphic DNA and nucleotide sequences in a region of the 18S rRNA and the internal transcribed spacer 1. Isolate-specific primers for these two regions were designed, and PCR tests capable of discriminating between isolates were developed. In both PCR assays, the five C. parvum isolates analyzed segregated into two subgroups. One group consisted of isolates that originated directly from human patients, and the other group had various host origins and had been propagated in laboratory animals. These results demonstrate the feasibility of distinguishing C. parvum isolates by sequence-specific PCR tests.


MSRF-Net: A Multi-Scale Residual Fusion Network for Biomedical Image Segmentation.
Methods based on convolutional neural networks have improved the performance of biomedical image segmentation. However, most of these methods cannot efficiently segment objects of variable sizes and train on small and biased datasets, which are common for biomedical use cases. While methods exist that incorporate multi-scale fusion approaches to address the challenges arising with variable sizes, they usually use complex models that are more suitable for general semantic segmentation problems. In this paper, we propose a novel architecture called Multi-Scale Residual Fusion Network (MSRF-Net), which is specially designed for medical image segmentation. The proposed MSRF-Net is able to exchange multi-scale features of varying receptive fields using a Dual-Scale Dense Fusion (DSDF) block. Our DSDF block can exchange information rigorously across two different resolution scales, and our MSRF sub-network uses multiple DSDF blocks in sequence to perform multi-scale fusion. This allows the preservation of resolution, improved information flow and propagation of both high- and low-level features to obtain accurate segmentation maps. The proposed MSRF-Net allows to capture object variabilities and provides improved results on different biomedical datasets. Extensive experiments on MSRF-Net demonstrate that the proposed method outperforms the cutting-edge medical image segmentation methods on four publicly available datasets. We achieve the Dice Coefficient (DSC) of 0.9217, 0.9420, and 0.9224, 0.8824 on Kvasir-SEG, CVC-ClinicDB, 2018 Data Science Bowl dataset, and ISIC-2018 skin lesion segmentation challenge dataset respectively. We further conducted generalizability tests and achieved DSC of 0.7921 and 0.7575 on CVC-ClinicDB and Kvasir-SEG, respectively.


Cytoglobin conformations and disulfide bond formation.
The oligomeric state and kinetics of ligand binding were measured for wild-type cytoglobin. Cytoglobin has the classical globin fold, with an extension at each extremity of about 20 residues. The extended length of cytoglobin leads to an ambiguous interpretation of its oligomeric state. Although the hydrodynamic diameter corresponds to that of a dimer, it displays a mass of a single subunit, indicating a monomeric form. Thus, rather than displaying a compact globular form, cytoglobin behaves hydrodynamically like a tightly packed globin with a greater flexibility of the N- and C-terminal regions. Cytoglobin displays biphasic kinetics after the photolysis of CO, as a result of competition with an internal protein ligand, the E7 distal histidine. An internal disulfide bond may form which modifies the rate of dissociation of the distal histidine and apparently leads to different cytoglobin conformations, which may affect the observed oxygen affinity by an order of magnitude.


Gene therapy for hemophilia.
This review will highlight the progress achieved in the past 2 years on using gene therapy to treat hemophilia in animals and humans. There has been substantial progress in using gene therapy to treat animals with hemophilia. Novel approaches for hemophilia A in mice include expression of Factor VIII in blood cells or platelets derived from ex-vivo transduced hematopoietic stem cells, or in-vivo transfer of transposons expressing Factor VIII into endothelial cells or hepatocytes. Advances in large-animal models include the demonstration that neonatal administration of a retroviral vector expressing canine Factor VIII completely corrected hemophilia A in dogs, and that double-stranded adeno-associated virus vectors resulted in expression of Factor IX that is 28-fold that obtained using single-stranded adeno-associated virus vectors. In humans, one hemophilia B patient achieved 10% of normal activity after liver-directed gene therapy with a single-stranded adeno-associated virus vector expressing human Factor IX. Expression fell at 1 month, however, which was likely due to an immune response to the modified cells. Gene therapy has been successful in a patient with hemophilia B, but expression was unstable due to an immune response. Abrogating immune responses is the next major hurdle for achieving long-lasting gene therapy.


The RNAmute web server for the mutational analysis of RNA secondary structures.
RNA mutational analysis at the secondary-structure level can be useful to a wide-range of biological applications. It can be used to predict an optimal site for performing a nucleotide mutation at the single molecular level, as well as to analyze basic phenomena at the systems level. For the former, as more sequence modification experiments are performed that include site-directed mutagenesis to find and explore functional motifs in RNAs, a pre-processing step that helps guide in planning the experiment becomes vital. For the latter, mutations are generally accepted as a central mechanism by which evolution occurs, and mutational analysis relating to structure should gain a better understanding of system functionality and evolution. In the past several years, the program RNAmute that is structure based and relies on RNA secondary-structure prediction has been developed for assisting in RNA mutational analysis. It has been extended from single-point mutations to treat multiple-point mutations efficiently by initially calculating all suboptimal solutions, after which only the mutations that stabilize the suboptimal solutions and destabilize the optimal one are considered as candidates for being deleterious. The RNAmute web server for mutational analysis is available at http://www.cs.bgu.ac.il/~xrnamute/XRNAmute.


Macrophage migration inhibitory factor mediates protease-activated receptor 4-induced bladder pain through urothelial high mobility group box 1.
Macrophage migration inhibitory factor (MIF) mediates pain although the mechanisms are not well understood. Urothelial activation of protease activated receptor 4 (PAR4) results in urothelial MIF release, urothelial high mobility group box 1 (HMGB1) release and bladder pain in mice without bladder inflammation. All three effects are prevented by MIF inhibition while intravesical disulfide HMGB1 alone can induce bladder pain. This study utilizes genetic MIF deletion to determine whether MIF mediates PAR4-induced bladder pain and is upstream of HMGB1-induced bladder pain. Wild type (C57/BL6) and MIF knockout (KO) mice were treated with intravesical PAR4 activating peptide or disulfide HMGB1 and tested for abdominal mechanical hypersensitivity at baseline (before treatment) and 24 h after injection. Micturition parameters and bladder histology were examined after behavioral test. Real-time PCR and western blotting measured HMGB1 mRNA and protein levels in the bladders of naïve wild type and MIF KO mice, while immunofluorescence measured HMGB1 protein levels in the urothelium of both strains. Intravesical PAR4 activation resulted in abdominal mechanical hypersensitivity in wild-type mice but not MIF KO mice. Intravesical disulfide HMGB1 induced abdominal mechanical hypersensitivity in both strains. Neither treatment resulted in significant changes in micturition or bladder histology in either strain. HMGB1 mRNA and protein levels were higher in MIF KO mouse bladders and the urothelium of MIF KO bladder had greater immunostaining than the wild-type strain. MIF is a pivotal molecule mediating PAR4-induced bladder pain and regulating urothelial HMGB1 production and release to elicit bladder pain.


Determination of functional effects of mutations in the steroid 21-hydroxylase gene (CYP21) using recombinant vaccinia virus.
Steroid 21-hydroxylase (P450c21) is absent or defective in more than 90% of patients with congenital adrenal hyperplasia. This disorder of cortisol biosynthesis occurs in a wide spectrum of clinical severity; specific mutations in the 21-hydroxylase gene (CYP21) have been found in association with particular clinical phenotypes. To determine the functional effects of mutations causing amino acid substitutions, normal P450c21 and three mutagenized P450c21 enzymes were expressed at high levels in cultured COS-1 cells using recombinant vaccinia virus. A single amino acid substitution (Val281----Leu) present in patients with mild "nonclassical" 21-hydroxylase deficiency resulted in an enzyme with 20-50% of normal activity. A mutation (Ile172----Asn) identified in patients with the "simple virilizing" form (poor cortisol synthesis but adequate aldosterone synthesis) resulted in an enzyme with less than 2% of normal activity. Finally, a cluster mutation (Ile-Val-Glu-Met234-238----Asn-Glu-Glu-Lys) found in a patient with severe "salt wasting" 21-hydroxylase deficiency (inadequate aldosterone synthesis) results in an enzyme with no detectable activity. These data indicate that the severity of 21-hydroxylase deficiency correlates with the degree of enzymatic compromise.


Translational control by the La antigen. Structure requirements for rescue of the double-stranded RNA-mediated inhibition of protein synthesis.
The La antigen is a protein which can bind both single-stranded and double-stranded forms of RNA and has regulatory effects on gene expression at the levels of transcription and translation. It was previously shown to inhibit the activation of the dsRNA-dependent protein kinase PKR by sequestering and/or unwinding double-stranded RNA. Here, we demonstrate that, as predicted by these properties, the La antigen can rescue protein synthesis in the reticulocyte lysate system from inhibition by low concentrations of dsRNA. This effect is reversed by higher concentrations of dsRNA. Using a series of deletion mutants we have investigated the structural features of the La antigen that are required for these effects. The ability to bind dsRNA is influenced by regions within both the previously characterized N-terminal RNP motif and the C-terminal half of the protein. La mutants with either N-terminal or C-terminal deletions retain the ability to inhibit the protein kinase activity of PKR and to rescue protein synthesis from inhibition by dsRNA. It is notable that sequences in the C-terminal half of the La antigen, including a phosphorylation site at Ser366, which are needed for other regulatory effects of the protein on gene expression are dispensable for the effects of La on PKR. We suggest that La regulates PKR activity solely as a result of its ability to act as an RNA-binding protein that can compete with PKR for limiting amounts of dsRNA.


Drimane Sesquiterpene Aldehydes Control Candida Yeast Isolated from Candidemia in Chilean Patients.
Drimys winteri J.R. (Winteraceae) produce drimane sesquiterpenoids with activity against Candida yeast. In this work, drimenol, polygodial (1), isotadeonal (2), and a new drimane α,β-unsaturated 1,4-dialdehyde, named winterdial (4), were purified from barks of D. winteri. The oxidation of drimenol produced the monoaldehyde drimenal (3). These four aldehyde sesquiterpenoids were evaluated against six Candida species isolated from candidemia patients in Chilean hospitals. Results showed that 1 displays fungistatic activity against all yeasts (3.75 to 15.0 µg/mL), but irritant effects on eyes and skin, whereas its non-pungent epimer 2 has fungistatic and fungicide activities at 1.9 and 15.0 µg/mL, respectively. On the other hand, compounds 3 and 4 were less active. Molecular dynamics simulations suggested that compounds 1-4 are capable of binding to the catalytic pocket of lanosterol 14-alpha demethylase with similar binding free energies, thus suggesting a potential mechanism of action through the inhibition of ergosterol synthesis. According to our findings, compound 2 appears as a valuable molecular scaffold to pursue the future development of more potent drugs against candidiasis with fewer side effects than polygodial. These outcomes are significant to broaden the alternatives to treat fungal infections with increasing prevalence worldwide using natural compounds as a primary source for active compounds.


A case of drug-induced hypersensitivity syndrome induced by salazosulfapyridine combined with SIADH caused by interstitial pneumonia.
We present a case of a patient with drug-induced hypersensitivity syndrome (DIHS) caused by salazosulfapyridine combined with syndrome of inappropriate secretion of antidiuretic hormone (SIADH) caused by interstitial pneumonia (IP). A 67-year-old man with a past history of rheumatism (RA) presented with right hemiparalysis and aphasia as the chief complaints. A diagnosis of left embolic cerebral infarction following trial therapy for RA based on computed tomography findings was made, and external decompression was performed. Salazosulfapyridine was newly started on day 7. Dabigatran was started on day 37. On day 41, the patient developed fever. On day 42, edema and erythema appeared on his face, and erythema and rash appeared on his trunk and extremities, with gradual transition to erythroderma. The drug eruption was initially attributed to the dabigatran. Various symptoms of organ dysfunction (enteritis, myocarditis, interstitial pneumonia, hepatic disorder, stomatitis, and others) then appeared and persisted; hence, a diagnosis of DIHS associated with human herpes virus 6 and cytomegalovirus infection induced by salazosulfapyridine was suggested, and the oral administration of salazosulfapyridine was discontinued on day 53. Hyponatremia was observed in association with exacerbation of IP. Due to low serum osmotic pressure and prompt improvement of the serum sodium level by fluid restriction, the SIADH was attributed to IP. In this case, steroid pulse therapy followed by gradual decrease therapy prevented worsening of the condition.