Genotyping microsatellite polymorphisms by agarose gel electrophoresis with ethidium bromide staining: application to quantitative trait loci analysis of seizure susceptibility in mice.
Agarose gel electrophoresis with ethidium bromide staining (AGE/EBS) is an efficient and reliable method for analyzing microsatellite polymorphisms. We report the use of AGE/EBS for analyzing DNA microsatellite polymorphisms in a preliminary quantitative trait loci (QTL) study of seizure susceptibility in which a candidate gene strategy was used to direct initial mapping efforts. F2 intercross progeny, derived from seizure-sensitive DBA/2J (D2) and seizure-resistant C57BL/6J (B6) inbred strains of mice, were tested for their sensitivity to the seizure-inducing effect of pentylenetetrazol (PTZ), a gamma-aminobutyric acid (GABA) receptor antagonist. A semi-automated method is described, in which DNA microsatellites were amplified by polymerase chain reaction (PCR) to yield products of 100-200 base pair (bp) in length. Alleles were separated on 3-6% MetaPhor agarose gels, stained with ethidium bromide, and visualized by ultraviolet (UV) illumination. Univariate analysis of genotype and phenotype data provides evidence for a seizure-related QTL on chromosome 5, near genes coding for the GABAA receptor subunits alpha 5 and gamma 3. Interestingly, this suggestive QTL derives from the more resistant B6 strain, but it nonetheless provides impetus for the characterization of possible strain differences in these two candidate genes. Overall, these results demonstrate that AGE/EBS can be useful for rapid screening of genomic regions of special interest in QTL mapping studies.


ANMCO POSITION PAPER: on administration of type 2 sodium-glucose co-transporter inhibitors to prevent heart failure in diabetic patients and to treat heart failure patients with and without diabetes.
This ANMCO (Associazione Nazionale Medici Cardiologi Ospedalieri) position paper aims to analyse the complex action of sodium-glucose co-transporter 2 inhibitors at the level of the kidney and cardiovascular system, focusing on the effect that these molecules have shown in the prevention and treatment of heart failure in diabetic and non-diabetic subjects. The goal was pursued by comparing the data generated with pathophysiology studies and with multicentre controlled studies in large populations. In accordance with the analysis carried out in the document, the following recommendations are issued: (i) canagliflozin, dapagliflozin, empagliflozin, and ertugliflozin are molecules recommended for the prevention of heart failure hospitalizations in type 2 diabetic subjects; (ii) canagliflozin and dapagliflozin are recommended for the prevention of heart failure hospitalizations in type 2 diabetic subjects with severe chronic kidney disease, dapagliflozin proved to be safe and effective also in diabetic subjects; and (iii) dapagliflozin and empagliflozin are recommended to reduce the combined risk of heart failure and cardiovascular death in diabetic and non-diabetic subjects with heart failure and reduced ejection fraction.


Aspirations and Worries: The Role of Parental Intrinsic Motivation in Establishing Oral Health Practices for Indigenous Children.
Aboriginal and Torres Strait Islander (respectfully, subsequently referred to as Indigenous) children in Australia experience oral disease at a higher rate than non-Indigenous children. A history of colonisation, government-enforced assimilation, racism, and cultural annihilation has had profound impacts on Indigenous health, reflected in oral health inequities sustained by Indigenous communities. Motivational interviewing was one of four components utilised in this project, which aimed to identify factors related to the increased occurrence of early childhood caries in Indigenous children. This qualitative analysis represents motivational interviews with 226 participants and explores parents' motivations for establishing oral health and nutrition practices for their children. Findings suggest that parental aspirations and worries underscored motivations to establish oral health and nutrition behaviours for children in this project. Within aspirations, parents desired for children to 'keep their teeth' and avoid false teeth, have a positive appearance, and preserve self-esteem. Parental worries related to child pain, negative appearance, sugar consumption, poor community oral health and rotten teeth. A discussion of findings results in the following recommendations: (1) consideration of the whole self, including mental health, in future oral health programming and research; (2) implementation of community-wide oral health programming, beyond parent-child dyads; and (3) prioritisation of community knowledge and traditions in oral health programming.


Humoral and haemodynamic interactions between clonidine and nifedipine in human essential hypertension.
Acute administration of nifedipine (NIF), a calcium entry blocker (CEB), in animals reduces the hypotensive effect of clonidine (CLN), an alpha-adrenergic agonist. In order to evaluate possible negative interactions between NIF and CLN in man during chronic treatment with these drugs, 12 patients with mild to moderate uncomplicated essential hypertension received either NIF (20 mg twice daily), CLN (0.25 mg once daily), the two drugs together at the same doses, or their matched placebos for a 2-week period each. NIF (-13.5% versus placebo) and CLN (-10.2% versus placebo) decreased blood pressure significantly and when combined, blood pressure was further decreased (-17.1% versus placebo). Heart rate was increased by NIF and NIF + CLN but unchanged by CLN alone. Plasma renin activity (PRA) tended to increase with NIF, decreased with CLN (P < 0.05) and was unchanged with NIF + CLN. Plasma aldosterone did not vary during any of the phases of the trial. These results indicate that NIF and CLN do not interact negatively on blood pressure control in essential hypertensive subjects.


Racial discrimination and child and adolescent health in longitudinal studies: A systematic review.
The association between racial discrimination and adverse health outcomes has been documented across an increasing body of evidence in recent years, although a close examination of longitudinal studies has not yet taken place. This review applied a life course lens in examining the evidence for a longitudinal association between racial discrimination exposure during childhood and adolescence, and later mental and physical health outcomes. Medline, PsycINFO, Global Health, ERIC, CINAHL Plus, Academic Search Premier and SocINDEX were searched from earliest records to October 2017 for eligible articles. Results were described through a narrative synthesis of the evidence. Findings from 46 studies reported in 88 empirical articles published between 2003 and 2017 were identified. Studies were primarily based on cohorts from the United States, comprised of young people aged 11-18 years, and were published since 2010. Data were most frequently collected over two to three timepoints at intervals exceeding 12 months. Statistically significant associations with racial discrimination were most commonly reported for behaviour problems including delinquency and risk-taking behaviour, with significant adverse effects found in 74% of these associations. Statistically significant adverse effects were also reported in 63% of associations with health-harming behaviours including substance use, and 61% found associations with mental health outcomes. Consistently significant associations were reported between accumulated racism and later health outcomes, and the health effects of racism were reported to vary with developmental periods, although few studies featured these analyses. Evidence from this review highlights that the duration and timing of exposure to racial discrimination matters. This review emphasises the need to gain evidence for the mechanisms linking early racism exposure to adverse health outcomes in later life. Future longitudinal research can address this need by capitalising on prospective cohort studies and ensuring that proposed analysis informs variable selection and timing of data collection.


The contribution of a complex systems-based approach to progressive social resilience.
The use of resilience in social practice has evolved from a theoretical framework at the intersection between individuals and their social ecology. Critics argue this theory still results in policies and practices that are too individualised, with the potential for negative social consequences. This paper further critiques contemporary understanding of resilience theory and its application. It juxtaposes complex systems theory with a social inequalities oriented resilience practice. This provides a paradoxical approach. It is acknowledged that state and public policy decisions and actions can be anti-resilient, undermining community and social resilience that already exists in the form of social relationships, self-organisation and co-production. Nevertheless, collective social resilience also illustrates the potential of local and service user organisations to contribute to an overall transformational change process.


The carboxyl-terminal domain of phosphophoryn contains unique extended triplet amino acid repeat sequences forming ordered carboxyl-phosphate interaction ridges that may be essential in the biomineralization process.
Phosphophoryns (PPs), a family of Asp and Ser(P)-rich dentin proteins, are considered to be archetypal regulators of several aspects of extracellular matrix (ECM) biomineralization. We have cloned a rat incisor PP gene, Dmp2, from our odontoblast cDNA library and localized it to mouse chromosome 5q21 within 2 centimorgans of Dmp1, another tooth-specific ECM protein. The carboxyl-terminal region of Dmp2 protein (60 residue % Ser, 31 residue % Asp) is divided into two domains, one with unique repetitive blocks of [DSS]n,3</=14, the other with [SD]m = 2,3. Conformational analysis shows the phosphorylated form of the [DS*S*]n repeats to have a unique structure with well defined ridges of phosphates and carboxyls available for counter ion binding. The [S*D]m domains have different phosphate and carboxylate interaction edges and thus different calcium ion and apatite surface binding properties. These two domains and the colocalization of Dmp1 and Dmp2 genes at a position equivalent to the dentinogenesis imperfecta type II location on human 4q21 all suggest that the PPs are indeed involved in some aspect of ECM mineralization.


The rice CK2 kinase regulates trafficking of phosphate transporters in response to phosphate levels.
Phosphate transporters (PTs) mediate phosphorus uptake and are regulated at the transcriptional and posttranslational levels. In one key mechanism of posttranslational regulation, phosphorylation of PTs affects their trafficking from the endoplasmic reticulum (ER) to the plasma membrane. However, the kinase(s) mediating PT phosphorylation and the mechanism leading to ER retention of phosphorylated PTs remain unclear. In this study, we identified a rice (Oryza sativa) kinase subunit, CK2β3, which interacts with PT2 and PT8 in a yeast two-hybrid screen. Also, the CK2α3/β3 holoenzyme phosphorylates PT8 under phosphate-sufficient conditions. This phosphorylation inhibited the interaction of PT8 with PHOSPHATE TRANSPORTER TRAFFIC FACILITATOR1, a key cofactor regulating the exit of PTs from the ER to the plasma membrane. Additionally, phosphorus starvation promoted CK2β3 degradation, relieving the negative regulation of PT phosphorus-insufficient conditions. In accordance, transgenic expression of a nonphosphorylatable version of OsPT8 resulted in elevated levels of that protein at the plasma membrane and enhanced phosphorus accumulation and plant growth under various phosphorus regimes. Taken together, these results indicate that CK2α3/β3 negatively regulates PTs and phosphorus status regulates CK2α3/β3.


NGF-induced axon growth is mediated by localized inactivation of GSK-3beta and functions of the microtubule plus end binding protein APC.
Little is known about how nerve growth factor (NGF) signaling controls the regulated assembly of microtubules that underlies axon growth. Here we demonstrate that a tightly regulated and localized activation of phosphatidylinositol 3-kinase (PI3K) at the growth cone is essential for rapid axon growth induced by NGF. This spatially activated PI3K signaling is conveyed downstream through a localized inactivation of glycogen synthase kinase 3beta (GSK-3beta). These two spatially coupled kinases control axon growth via regulation of a microtubule plus end binding protein, adenomatous polyposis coli (APC). Our results demonstrate that NGF signals are transduced to the axon cytoskeleton via activation of a conserved cell polarity signaling pathway.


Electrochemical Properties of Sulfurized-Polyacrylonitrile Cathode for Lithium-Sulfur Batteries: Effect of Polyacrylic Acid Binder and Fluoroethylene Carbonate Additive.
Sulfurized carbonized polyacrylonitrile (S-CPAN) is a promising cathode material for Li-S batteries owing to the absence of polysulfide dissolution phenomena in the electrolyte solutions and thus the lack of a detrimental shuttle mechanism. However, challenges remain in achieving high performance at practical loading because of large volume expansion of S-CPAN electrodes and lithium anode degradation at high current densities. To mitigate this problem, we propose a novel cell design including poly(acrylic acid) (PAA) binder for improved integrity of the composite electrodes and fluoroethylene carbonate (FEC) as additive in the electrolyte solutions for stabilizing the lithium metal surface. As a result, these cells delivered high initial discharge capacity of 1500 mAh g-1 and a superior cycling stability ∼98.5% capacity retention after 100 cycles, 0.5 C rate, and high sulfur loading of 3.0 mg cm-2. Scaled-up 260 mAh pouch cells are working very well, highlighting the practical importance of this work.


Role of monocyte/macrophage population in immune response.
The central role of macrophages in host defence against infection and malignancy and processes such as atherosclerosis, makes macrophage biology a fascinating area for research in immunology and cell biology. The endocytic and phagocytic machinery of macrophages is particularly potent and their secretory potential is large and diverse. Studies of cell surface receptors and their role in antigen presentation, microbicidal and tumouricidal activity are actively researched and progress is now being made in defining receptors responsible for monocyte/macrophage cell adhesion within the immune system. This short-review highlights the recent advances in macrophage biology.


Infections after craniotomy: a retrospective study.
A retrospective study was performed to evaluate the rate of infection directly related to craniotomy. Of 246 craniotomies performed in 1981, 234 could be evaluated. The overall incidence of infection was 8.1% (19 infections in 18 patients). The infection rate was 15.2% after insertion of a ventricular shunt (seven infections after 45 ventriculostomies). Staphylococcus aureus, Staphylococcus epidermidis and Propionibacterium acnes were the most frequent isolated micro-organisms. We recommended a double-blind prospective study to evaluate the possible benefit of antibiotic prophylaxis in craniotomies.


METTL3 regulates viral m6A RNA modification and host cell innate immune responses during SARS-CoV-2 infection.
It is urgent and important to understand the relationship of the widespread severe acute respiratory syndrome coronavirus clade 2 (SARS-CoV-2) with host immune response and study the underlining molecular mechanism. N6-methylation of adenosine (m6A) in RNA regulates many physiological and disease processes. Here, we investigate m6A modification of the SARS-CoV-2 gene in regulating the host cell innate immune response. Our data show that the SARS-CoV-2 virus has m6A modifications that are enriched in the 3' end of the viral genome. We find that depletion of the host cell m6A methyltransferase METTL3 decreases m6A levels in SARS-CoV-2 and host genes, and m6A reduction in viral RNA increases RIG-I binding and subsequently enhances the downstream innate immune signaling pathway and inflammatory gene expression. METTL3 expression is reduced and inflammatory genes are induced in patients with severe coronavirus disease 2019 (COVID-19). These findings will aid in the understanding of COVID-19 pathogenesis and the design of future studies regulating innate immunity for COVID-19 treatment.


Quantitative electroencephalograms and neuro-optometry: a case study that explores changes in electrophysiology while wearing therapeutic eyeglasses.
The brain is equipped with a complex system for processing sensory information, including retinal circuitry comprising part of the central nervous system. Retinal stimulation can influence brain function via customized eyeglasses at both subcortical and cortical levels. We investigated cortical effects from wearing therapeutic eyeglasses, hypothesizing that they can create measureable changes in electroencephalogram (EEG) tracings. A Z-BellSM test was performed on a participant to select optimal lenses. An EEG measurement was recorded before and after the participant wore the eyeglasses. Equivalent quantitative electroencephalography (QEEG) analyses (statistical analysis on raw EEG recordings) were performed and compared with baseline findings. With glasses on, the participant's readings were found to be closer to the normed database. The original objective of our investigation was met, and additional findings were revealed. The Z-bellSM test identified lenses to influence neurotypical brain activity, supporting the paradigm that eyeglasses can be utilized as a therapeutic intervention. Also, EEG analysis demonstrated that encephalographic techniques can be used to identify channels through which neuro-optomertric treatments work. This case study's preliminary exploration illustrates the potential role of QEEG analysis and EEG-derived brain imaging in neuro-optometric research endeavors to affect brain function.


Osteoinductive Electrospun Scaffold Based on PCL-Col as a Regenerative Therapy for Peri-Implantitis.
Peri-implantitis is a serious condition affecting dental implants that can lead to implant failure and loss of osteointegration if is not diagnosed and treated promptly. Therefore, the development of new materials and approaches to treat this condition is of great interest. In this study, we aimed to develop an electrospun scaffold composed of polycaprolactone (PCL) microfibers loaded with cholecalciferol (Col), which has been shown to promote bone tissue regeneration. The physical and chemical properties of the scaffold were characterized, and its ability to support the attachment and proliferation of MG-63 osteoblast-like cells was evaluated. Our results showed that the electrospun PCL-Col scaffold had a highly porous structure and good mechanical properties. The resulting scaffolds had an average fiber diameter of 2-9 μm and high elongation at break (near six-fold under dry conditions) and elasticity (Young modulus between 0.9 and 9 MPa under dry conditions). Furthermore, the Col-loaded scaffold was found to decrease cell proliferation when the Col content in the scaffolds increased. However, cytotoxicity analysis proved that the PCL scaffold on its own releases more lactate dehydrogenase into the medium than the scaffold containing Col at lower concentrations (PCL-Col A, PCL-Col B, and PCL-Col C). Additionally, the Col-loaded scaffold was shown to effectively promote the expression of alkaline phosphatase and additionally increase the calcium fixation in MG-63 cells. Our findings suggest that the electrospun membrane loaded with Col can potentially treat peri-implantitis by promoting bone formation. However, further studies are needed to assess the efficacy and safety of this membrane in vivo.


Partial purification and characterization of dynein adenosine triphosphatase from bovine sperm.
Outer dynein arm polypeptides that possess Mg+2-adenosine triphosphatase (ATPase) activity have been extracted from the flagellar axonemes of demembranated bovine sperm. Electron microscopy of intact and salt-extracted sperm demonstrates a relatively selective removal of the outer dynein arms. The salt extract contains a specific ATPase activity of 55 nmoles inorganic phosphate (Pi)/min/mg protein. Sucrose density gradient centrifugation of this extract results in a 6-fold increase in specific activity of ATPase (333 nmole/Pi/min/mg protein), which sediments as a single 13S peak. Concomitant with the increase in specific activity, there is enrichment of three high molecular weight polypeptides (Mr greater than 300,000) characteristic of dynein heavy chains. ATPase activities in the initial extract and in the 13S peak are inhibited by concentrations of vanadate and erythro-9-[3-2-(hydroxynonyl)]adenine similar to those that inhibit ATPase activity in sea urchin sperm dynein. These findings indicate that outer arm dynein ATPase can be extracted and partially purified from bovine sperm.


Content Factor: a measure of a journal's contribution to knowledge.
Impact Factor, the pre-eminent performance metric for medical journals, has been criticized for failing to capture the true impact of articles; for favoring methodology papers; for being unduly influenced by statistical outliers; and for examining a period of time too short to capture an article's long-term importance. Also, in the era of search engines, where readers need not skim through journals to find information, Impact Factor's emphasis on citation efficiency may be misplaced. A better metric would consider the total number of citations to all papers published by the journal (not just the recent ones), and would not be decremented by the total number of papers published. We propose a metric embodying these principles, "Content Factor", and examine its performance among leading medical and orthopaedic surgery journals. To remedy Impact Factor's emphasis on recent citations, Content Factor considers the total number of citations, regardless of the year in which the cited paper was published. To correct for Impact Factor's emphasis on efficiency, no denominator is employed. Content Factor is thus the total number of citations in a given year to all of the papers previously published in the journal. We found that Content Factor and Impact Factor are poorly correlated. We further surveyed 75 experienced orthopaedic authors and measured their perceptions of the "importance" of various orthopaedic surgery journals. The correlation between the importance score and the Impact Factor was only 0.08; the correlation between the importance score and Content Factor was 0.56. Accordingly, Content Factor better reflects a journal's "importance". In sum, while Content Factor cannot be defended as the lone metric of merit, to the extent that performance data informs journal evaluations, Content Factor--an easily obtained and intuitively appealing metric of the journal's knowledge contribution, not subject to gaming--can be a useful adjunct.


Longitudinal effects of menopausal hormone treatments on platelet characteristics and cell-derived microvesicles.
Activated platelets serve as a catalyst for thrombin generation and a source of vasoactive and mitogenic factors affecting vascular remodeling. Oral menopausal hormone treatments (MHT) may carry greater thrombotic risk than transdermal products. This study compared effects of oral and transdermal MHT on platelet characteristics, platelet proteins, and platelet-derived microvesicles (MV) in recently menopausal women. Platelets and MV were prepared from blood of a subset of women (n = 117) enrolled in the Kronos Early Estrogen Prevention Study prior to and after 48 months of treatment with either oral conjugated equine estrogen (0.45 mg/day), transdermal 17β-estradiol (50 µg/day), each with intermittent progesterone (200 mg/day for 12 days a month), or placebo pills and patch. Platelet count and expression of platelet P-selectin and fibrinogen receptors were similar across groups. An aggregate measure of 4-year change in vasoactive and mitogenic factors in platelet lysate, by principle component analysis, indicated significantly lower values in both MHT groups compared to placebo. Increases in numbers of tissue factor positive and platelet-derived MV were significantly greater in the transdermal compared to placebo group. MHT was associated with significantly reduced platelet content of vasoactive and mitogenic factors representing a potential mechanism by which MHT may affect vascular remodeling. Various hormonal compositions and doses of MHT could differentially regulate nuclear transcription in bone marrow megakaryocytes and non-genomic pathways in circulating platelets thus determining numbers and characteristics of circulating MV. Thrombotic risk associated with oral MHT most likely involves liver-derived inflammatory/coagulation proteins rather than circulating platelets per se.


Tunable liquid crystal q-plates with arbitrary topological charge.
Using a photoalignment technique with a sulphonic azo-dye as the surfactant aligning material, we fabricated electrically tunable liquid crystal q-plates with topological charge 0.5, 1.5 and 3 for generating optical vortex beams with definite orbital angular momentum (OAM) 1,3 and 6 per photon (in units of ¯h), respectively. We carried out several tests on our q-plates, including OAM tomography, finding excellent performances. These devices can have useful applications in general and quantum optics.


Asymmetric chromatid segregation in cardiac progenitor cells is enhanced by Pim-1 kinase.
Cardiac progenitor cells (CPCs) in the adult heart are used for cell-based treatment of myocardial damage, but factors determining stemness, self-renewal, and lineage commitment are poorly understood. Immortal DNA strands inherited through asymmetric chromatid segregation correlate with self-renewal of adult stem cells, but the capacity of CPCs for asymmetric segregation to retain immortal strands is unknown. Cardioprotective kinase Pim-1 increases asymmetric cell division in vivo, but the ability of Pim-1 to enhance asymmetric chromatid segregation is unknown. We aimed to demonstrate immortal strand segregation in CPCs and the enhancement of asymmetric chromatid distribution by Pim-1 kinase. Asymmetric segregation is tracked by incorporation of bromodeoxyuridine. The CPC DNA was labeled for several generations and then blocked in second cytokinesis during chase to determine distribution of immortal versus newly synthesized strands. Intensity ratios of binucleated CPCs with bromodeoxyuridine of ≥70:30 between daughter nuclei indicative of asymmetric chromatid segregation occur with a frequency of 4.57, and asymmetric chromatid segregation is demonstrated at late mitotic phases. Asymmetric chromatid segregation is significantly enhanced by Pim-1 overexpression in CPCs (9.19 versus 4.79 in eGFP-expressing cells; P=0.006). Asymmetric segregation of chromatids in CPCs is increased nearly two-fold with Pim-1 kinase overexpression, indicating that Pim-1 promotes self-renewal of stem cells.


Corneal cross-linking in 9 horses with ulcerative keratitis.
Corneal ulcers are one of the most common eye problems in the horse and can cause varying degrees of visual impairment. Secondary infection and protease activity causing melting of the corneal stroma are always concerns in patients with corneal ulcers. Corneal collagen cross-linking (CXL), induced by illumination of the corneal stroma with ultraviolet light (UVA) after instillation of riboflavin (vitamin B2) eye drops, introduces crosslinks which stabilize melting corneas, and has been used to successfully treat infectious ulcerative keratitis in human patients. Therefore we decided to study if CXL can be performed in sedated, standing horses with ulcerative keratitis with or without stromal melting. Nine horses, aged 1 month to 16 years (median 5 years) were treated with a combination of CXL and medical therapy. Two horses were diagnosed with mycotic, 5 with bacterial and 2 with aseptic ulcerative keratitis. A modified Dresden-protocol for CXL could readily be performed in all 9 horses after sedation. Stromal melting, diagnosed in 4 horses, stopped within 24 h. Eight of nine eyes became fluorescein negative in 13.5 days (median time; range 4-26 days) days after CXL. One horse developed a bacterial conjunctivitis the day after CXL, which was successfully treated with topical antibiotics. One horse with fungal ulcerative keratitis and severe uveitis was enucleated 4 days after treatment due to panophthalmitis. CXL can be performed in standing, sedated horses. We did not observe any deleterious effects attributed to riboflavin or UVA irradiation per se during the follow-up, neither in horses with infectious nor aseptic ulcerative keratitis. These data support that CXL can be performed in the standing horse, but further studies are required to compare CXL to conventional medical treatment in equine keratitis and to optimize the CXL protocol in this species.


Nasopalatine duct cyst: report of 22 cases and review of the literature.
Nasopalatine duct cysts (NPDCs) are the most common developmental, epithelial and non-odontogenic cysts of the maxillae. The present study describes the clinicopathological characteristics of 22 NPDCs and discusses their etiology, incidence, treatment and prognosis, with a review of the literature on the subject. A retrospective observational study was made comprising a period of 36 years (1970-2006), and yielding a series of 22 patients with histopathological confirmation of NPDC. Surgical treatment was carried out under local anesthesia and comprised the dissection and removal of the cyst adopting a usually palatine approach, with the preparation of an enveloping flap from 1.4 to 2.4. No statistically significant correlation was observed between the size of the lesion and patient age, although the size of the cyst differed according to patient gender, with a mean NPDC diameter of 16 mm in males and 12 mm in females. In no case did we observe root resorption or loss of vitality of the upper incisors following surgery. The X-ray image was rounded in 15 cases and heart-shaped in the remaining 7 cases. In the majority of cases panoramic X-rays and periapical and occlusal X-rays sufficed to identify the lesion, though computed tomography was used in cases of doubt. The etiology of NPDC is unclear. Simple surgical resection is recommended, followed by clinical and radiological control to ensure correct resolution of the case.


Loading sequence is a major determinant of afterload-dependent relaxation in intact canine heart.
To elucidate the role of loading sequence in afterload-dependent slowed relaxation in hearts in situ, the time constants (Texp from best exponential fitting method and TL from semilogarithmic method) of isovolumetric left ventricular (LV) pressure decay were studied in nine anesthetized open-chest dogs under the pharmacological blockade of autonomic nerve activity. An afterload change was imposed by clamping the ascending or descending aorta to make the peak LV pressure early or late in systole. During afterload interventions, in contractions with the peak LV pressure in late systole Texp and TL were significantly (P less than 0.05) larger than in those with the peak LV pressure in early systole in any comparable peak LV pressure range. Moreover, both time constants were directly correlated (P less than 0.01) with the time of peak LV pressure irrespective of peak LV pressure and clamp mode of aorta. In another protocol, marked differences both in Texp and TL were also observed between each of 25 pairs of contractions with different loading sequence but with comparable peak LV pressure and LV dimension (segment length). Thus afterload-dependent slowed relaxation in hearts in situ could not be attributed to an increased total load but to the altered loading sequence associated with an increase in afterload.


The adenovirus E3 14.7-kilodalton protein which inhibits cytolysis by tumor necrosis factor increases the virulence of vaccinia virus in a murine pneumonia model.
The 14.7-kilodalton protein (14.7K protein) encoded by the adenovirus (Ad) E3 region inhibits tumor necrosis factor alpha (TNF-alpha)-mediated lysis of cells in tissue culture experiments, but the relevance of this effect in vivo is incompletely understood. To examine the effect of the ability of the Ad 14.7K protein to block TNF lysis upon viral pathogenesis in a murine model, we cloned the 14.7K protein-encoding gene into vaccinia virus (VV), permitting its study in isolation from other Ad E3 immunomodulatory proteins. The gene for murine TNF-alpha was inserted into the same VV containing the 14.7K gene to ensure that each cell infected with the VV recombinant would express both the agonist (TNF) and its antagonist (14.7K). VV was utilized as the vector because it accommodates large and multiple inserts of foreign DNA with faithful, high-level expression of the protein products. In addition, infection of mice with VV induces disease with quantifiable morbidity, mortality, and virus replication. The results of intranasal infections of BALB/c mice with these VV recombinants indicate that the Ad 14.7K protein increases the virulence of VV carrying the TNF-alpha gene by reversing the attenuating effect of TNF-alpha on VV pathogenicity. This was demonstrated by increased mortality, pulmonary pathology, and viral titers in lung tissue following infection with VV coexpressing the 14.7K protein and TNF-alpha, compared with the control virus expressing TNF-alpha alone. These results suggest that the 14.7K protein, which is nonessential for Ad replication in tissue culture, is an immunoregulatory protein which functions in vivo to help counteract the antiviral effects of TNF-alpha.


Envy's pathology: Historical contexts.
This article is concerned with the physicality of envy primarily in early -modern, but also in eighteenth-century health contexts. The discussion brings together descriptions of the effects of envy on the body of the envier, mainly from works of physiology and health preservation, but also from literary and spiritual writings. These depictions of envy are studied beyond their symbolism and with a view to establish whether they are meaningful according to the medical theories of the time in which they occur. The discussion begins by acknowledging the status of envy as a 'disease' and looks to the specific ways in which the discourse of envy conveys this sense. I find that in the early modern discourse envy is always pathological, that is, it is experienced as disease and signifies disease in general and several diseases in particular. Moreover, envy is uniquely placed to convey pathology on account of its being connected to inherently pathogenic elements of the humoural theory. Specifically, envy is physiologically connected to melancholy, and the way it is presented comes close to attributes assigned to black bile. In addition, envy realizes pathology, the occurrence of disease in the body, by impairing the vital process of digestion and thus depriving the person from proper nourishment and sustenance. The analysis further considers how this impairment of the body fits with the physiological manifestation of envy as 'corrosion' and 'consumption'. Finding commonalities with other maladies mediated by these physiological signs the article concludes by considering the function of pathology in the conception of early modern envy.


Candida albicans dampens host defense by downregulating IL-17 production.
IL-17 is one of the key cytokines that stimulate host defense during a Candida infection. Several studies have demonstrated the capacity of Candida albicans to induce a Th17 response. Surprisingly, experiments employing live C. ablicans demonstrated a specific downregulation of host IL-17 secretion in human blood mononuclear cells (PBMCs). By avoiding the direct contact of live C. albicans and PBMCs, we demonstrate that this inhibition effect is mediated by a soluble factor released by live C. albicans. However, this effect is due neither to the releasing of C. albicans pathogen-associated molecular patterns nor to the alteration of different Th cell subtypes. Rather, we found that live C. albicans shifts tryptophan metabolism by inhibiting IDO expression away from kynurenines and toward 5-hydroxytryptophan metabolites. In addition, we show that these latter 5-hydroxytryptophan metabolites inhibit IL-17 production. In conclusion, live C. albicans inhibits host Th17 responses by modulatory effects on tryptophan metabolism.


Analysis of the spectrotypes of autoantibodies against thyroglobulin in two rat models of autoimmune thyroiditis.
We have studied the isoelectric focusing (IEF) spectrotypes of autoantibodies against thyroglobulin in two rat models of Hashimoto's thyroiditis: (1) AUG strain rats immunized with autologous thyroglobulin in Freund's complete adjuvant; (2) PVG/c strain rats which have been thymectomized and sublethally irradiated. The IEF spectrotypes revealed differences between the two models. The anti-thyroglobulin response in immunized AUG rats is mainly oligoclonal or polyclonal, often with dominant clones in the spectrotype, similar to about 90% of Hashimoto's patients, whereas the response in the PVG/c rat is highly restricted. There were pronounced changes in spectrotype with time in the PVG/c, but not AUG, rats with considerable variation in the lifespan of individual clones. The maximum lifespan of an anti-self secreting B-cell clone in the PVG/c rat, determined by persistence of its clonotype, was at least 16 weeks.


Vasopressin antagonists in the management of heart failure.
Vasopressin antagonists have been studied in a variety of clinical settings, including patients with acute and chronic heart failure. The clinical trials published to date have sought to describe the clinical and physiologic effects of these agents in an effort to prove clinical efficacy and safety. A variety of agents with varying effects on V2 and V1a vasopressin receptor subtype have been studied. They have been shown to reduce bodyweight and improve serum sodium without worsening renal function. They may also decrease the need for loop diuretic use and may be particularly useful in patients with hyponatremia in the setting of volume overload. Further studies are underway that are powered to assess for morbidity and mortality benefits. The beneficial effects have been well documented but, until outcomes are understood more fully, the use of these agents should be limited to currently approved indications. In the USA, this includes only the treatment of euvolemic hyponatremia.


Histomorphological and clinical study of primary and secondary glomerulopathies in Southeast Serbia (20-year period of analysis).
Epidemiological studies of renal biopsies have been performed to follow up the incidence of glomerular diseases on a specified territory and to compare the obtained results with results from other regions. The aim of this study was to analyze the frequency of certain histopathophysiological types of glomerular diseases on the territory of Southeast Serbia. In a 20-year period (1986-2006), 316 kidney biopsies were performed in patients with clinical signs of impaired renal function, in Southeast Serbia. On average 1.6 biopsies were made per year per 100 000 inhabitants. Biopsies of adult patients represented 88% of all biopsies, biopsies in children (aged under 18 years) represented 8%, while biopsies of elderly patients (more than 60 years) represented 4% of all biopsies. The predominance of male patients was described with male/female ratio of 1.4. The most frequent clinical manifestation in patients at the time of biopsy were nephrotic syndrome (42.5%), and asymptomatic proteinuria and/or hemamuria (31.3%) and nephritic syndrome (14.9%). The most common glomerular disease was IgA nephropathy with an incidence of 21.5% of total biopsy diagnosed glomerulopathies, followed by: membranous glomerulonephritis (12.6%), focal segmental proliferative and sclerosing glomerulonephritis (10.7%), lupus nephritis (8.4%), nephroangiosclerosis (7.0%), mesangio-proliferative glomerulonephritis (6.1%), minimal change disease (2.8%), mesangiocapillary glomerulonephritis (2.3%). The frequency of certain histopathologic findings significantly correlated with data from studies that we used for comparison, with the exception of minimal change disease whose incidence in our study was smaller.


Functional and conformational characterization of new mutants of heart fatty acid-binding protein.
In this study we investigated the possible involvement of several amino acids (not located in the ligand-binding centre) in fatty acid binding and conformational stability of heart fatty acid-binding protein (H-FABP). We prepared recombinant human H-FABP proteins with mutations in the hydrophobic patch (Phe(4), Trp(8) and Phe(64)), portal region (Phe(16)), hinge region (Leu(66), Gly(67)), second portal region (Glu(72)) and at the protein surface (Lys(21)) respectively. Oleic acid-binding affinity and conformational stability of human H-FABP are significantly decreased or completely lost by mutation of Trp(8) or Phe(16). NMR spectra confirmed that these residues are important for the stability of the protein fold. Substitution of Phe(4) or Phe(64) resulted in less stability, but oleic acid-binding affinity was not affected. Mutation of Lys(21) had no effect on either structural integrity or fatty acid-binding affinity. Replacement of Leu(66) or Gly(67) did not affect fatty acid binding, but protein stability was reduced. Finally, mutation of Glu(72) to Ser caused no change of affinity, but NMR spectra and urea-denaturation curves showed the extremely poor stability of this mutant. In conclusion, no relationship was observed between fatty acid-binding affinity and conformational stability.


The educational, vocational, and marital status of growth hormone-deficient adults treated with growth hormone during childhood.
The goal of growth hormone therapy in childhood is to increase stature, thereby facilitating normal psychosocial development. To determine the social outcome of patients with growth hormone deficiency (GHD), we interviewed 116 adults with GHD across Canada, including 86 men and 30 women 18 to 38 years of age who were treated with growth hormone during childhood. The education of the 96 patients who had completed their formal education was similar to their siblings and to the general population. Of the patients in the labor force, 35.4% were unemployed; the unemployment rates for those patients less than 25 years of age and those 25 years of age or older were 45% and 23%, respectively, compared with national rates of 21.2% and 9.4% for the same age groups, respectively. Of the 90 patients with GHD who were not attending school, 70 lived with their parents or relatives. Only 15 patients were married; one was divorced. The percentage of patients with GHD who were married was less than 30% of the expected age-adjusted rate. No difference in the rate of employment or marriage was found between the patients with idiopathic isolated GHD and organic hypopituitarism. In summary, the achievements of patients with GHD seem to be normal in the education system, but the rate of employment and marriage are much lower than expected. This poor outcome was unrelated to the response to growth hormone therapy and emphasizes the need to develop strategies that lead to more satisfactory psychosocial integration of patients with GHD in adult life.


[A comparative immunochemical study of the subtypes of serologic type I of the agent of pseudotuberculosis].
It was shown that Y. pseudotuberculosis strains causing the Far-Eastern scarlatina-like fever in the Primorsk region belonged to subtype IB-ipopolysaccharides of the standard strain of subtype IB and of the local strain were closely affiliated by the analytic data and monosaccharide composition, but differed from the lipopolysaccharide of strains belonging to subtype IA. Living vaccines should be used to obtain the sera against the subtypes IA and IB of the causative agent.


Effects of a Complexity-Based Approach on Generalization of Past Tense -ed and Related Morphemes.
In a previous article, we reported that beginning treatment for regular past tense -ed with certain types of verbs led to greater generalization in children with developmental language disorder than beginning treatment with other types of verbs. This article provides updated data from that study, including the addition of data from 3 children, results from naturalistic language samples, and data from a third time point. Twenty 4- to 9-year-old children with developmental language disorder (10 per condition) were randomly assigned to receive language intervention in which the verbs used to teach regular past tense -ed were manipulated. Half received easy first intervention, beginning with highly frequent, telic, phonologically simple verbs, and half received hard first intervention, beginning with less frequent, atelic, and phonologically complex verbs. The design used a train-to-criterion approach, with children receiving up to 36 visits. Performance was assessed using elicited production probes and language samples before intervention, immediately following intervention and 6-8 weeks later. Children in the hard first group showed greater gains on the use of regular past tense -ed in both structured probes (at immediate post only) and in language samples (at both immediate and delayed post). Gains attributable to therapy were not observed in untreated morphemes. This study suggests that the choice of therapy materials, with an eye on the role that treatment stimuli play in generalization, is important for treatment efficacy. Clinicians should consider early selection of atelic, lower-frequency, phonologically complex verbs when teaching children to use regular past tense -ed. Further work expanding this to other morphemes and a larger population is needed to confirm this finding.


[Immune adherence (C3b) receptor activity of erythrocytes in patients with esophageal cancer and cancer of the gastric cardia].
The red cell C3b receptor activity of 52 patients with esophageal cancer, 19 patients with cancer of gastric cardia and 31 age-matched normal persons was studied by the ability of forming ZC3b-rosette (rosette formation with zymosan particles treated by guinea pig complement) and IC-Z rosette (rosette formation with zymosan particles). The results showed that the ZC3b-rosette formation rate was significantly lower in cancer patients than that of the normal subjects (P less than 0.01), but the IC-Z rosette formation rate was significantly higher in the cancer patients (P less than 0.01). This suggests that the esophageal cancer and cancer of gastric cardia be associated with surface changes in the erythrocytes, especially those of the receptors which are responsible for immune adherence reaction. The methods of detecting the red cell C3b receptor are also described.


Directed self-assembly of peptide-diketopyrrolopyrrole conjugates - a platform for bio-organic thin film preparation.
Increased water solubility and long-range intermolecular ordering have been introduced into the fluorescent organic molecule thiophene-diketopyrrolopyrrole (TDPP) via its conjugation to the octapeptide HEFISTAH, which is derived from the protein-protein β-interface of the homo-tetramer protein diaminopimelate decarboxylase. The octapeptide, and its TDPP mono- and cross-linked conjugates were synthesised using 9-fluorenylmethoxycarbonyl (Fmoc) based solid-phase peptide synthesis (SPPS). Unlike the unmodified peptide, the resulting mono-linked and cross-linked peptides showed a fibrous morphology and formed hydrogels at 4 wt% in water at neutral pH, but failed to assemble at pH 2 and pH 9. Further peptide characterization showed that the TDPP organic core enhances peptide self-assembly and that both peptides assembled into fibers with a parallel β-sheet structure. Furthermore, UV-vis spectroscopic analysis suggests that the TDPP molecules form H-type aggregates where the chromophores are likely to be co-facially packed, but rotationally and/or laterally offset from one another. This intermolecular coupling indicates that π-π stacking interactions are highly likely - a favourable sign for charge transport. The enhanced aqueous solubility and self-assembling properties of the TDPP-peptide conjugates allowed the successful preparation of thin films. Atomic force microscopy, X-ray diffraction and UV-vis spectroscopic analysis of these thin films revealed that the hybrid materials retained a fibrous morphology, β-sheet structures and strong intermolecular coupling between neighbouring TDPP molecules. These results open an exciting avenue for bio-organic materials development, through structural and electronic tuning of the TDPP core.


Translational control analysis by translationally active RNA capture/microarray analysis (TrIP-Chip).
We have developed a new approach to systematically study post-transcriptional regulation in a small number of cells. Actively translating mRNAs are associated with polysomes and the newly synthesized peptide chains are closely associated with molecular chaperones such as hsp70s, which assist in the proper folding of nascent polypeptides into higher ordered structures. These chaperones provide an anchor with which to separate actively translating mRNAs associated with polysomes from free mRNAs. Affinity capture beads were developed to capture hsp70 chaperones associated with the polysome complexes. The isolated actively translating mRNAs were used for high-throughput expression profiling analysis. Feasibility was demonstrated using an in vitro translation system with known translationally regulated mRNA transcript thymidylate synthase (TS). We further developed the approach using HCT-116 colon cancer cells with both TS and p53 as positive controls. The steady-state levels of TS and p53 mRNAs were unaltered after 5-fluorouracil treatment as assessed by real-time qRT-PCR analysis. In contrast, the protein expression and polysome-associated mRNA levels of both genes were increased. These differences in translational rate were revealed with our new approach from 500 cells. This technology has the potential to make investigation of translational control feasible with limited quantities of clinical specimens.


Regulation of interferon-tau (IFN-tau) gene promoters by growth factors that target the Ets-2 composite enhancer: a possible model for maternal control of IFN-tau production by the conceptus during early pregnancy.
Expression of interferon-tau gene (IFNT) by conceptuses of cattle and sheep must be in phase with the physiological state of the mother if the pregnancy is to be successful. IFNT has a close-to-consensus AP1 site (-71 to -64), overlapping a binding site for Ets-2 (-79 to -70), the key transcription factor controlling IFNT expression. When a bovine IFNT gene control region-luciferase (luc) construct was transfected into mouse 3T3 fibroblasts in the presence of Ets-2 and oncogenic Ras, luc expression was activated (50- to 100-fold). Mutations in either the activator protein 1 (AP1) site or the Ets-2 site of this construct abolished this effect. Similarly, a mutation of Thr72 of the Ets-2 or the addition of a specific inhibitor for the MAPK signal transduction pathway also markedly reduced expression. IFNT promoter activity was up-regulated in response to colony-stimulating factor-1 in 3T3 cells expressing the colony-stimulating factor-1 receptor c-fms. This response did not occur when the AP1 site or the Ets-2 binding sites were mutated. Nor was the response observed in 3T3 cells expressing an inactive form of c-fms. The experiments indicate that IFNT can be activated by growth factors operating through the Ras/MAPK pathway. The Ets-2 and AP1 binding sites are essential for such effects. The AP1 site, however, is noncanonical and unable to bind either cJun or cFos. These data emphasize the importance of a complex Ets-2 enhancer for expression of IFNT and suggest a means whereby the mother can exert control over conceptus IFN-tau production.


New techniques and issues in assessing walking behavior and its contexts.
In the first section of this article, we discuss the metabolic responses to walking by describing the economy of walking during different locomotion velocities. Gender, weight status, and growth effects on metabolic responses to walking are reviewed. In the second section, we examine the use of technology to assess walking patterns and behavior in the community. We use an engineering approach for understanding how to measure objects that move, and these methods are used to assess walking used in transportation. In the third part of the paper, we summarize self-report methods that have been used to assess walking behavior and highlight the strengths and weaknesses of these methods. We illustrate how self-report methods are used to quantify walking behavior in the surveillance systems that are now widely used to ascertain walking prevalence and temporal changes in different populations. In the final section, we discuss ways of measuring the walkability of neighborhoods and the community to understand the influence of the built environment on walking behavior.


CT-simulator based brachytherapy planner: seed localization and incorporation of biological considerations.
Radiation dose prescription, interpretation, and planning can be problematic for brachytherapy due to high spatial heterogeneity, varying and various dose rates, absence of superimposed calculated isodose distributions onto affected tissues, and lack of dose volume histograms. A new treatment planner has been developed to reduce these limitations in brachytherapy planning. The PC-based planning system uses a CT-simulator to sequentially scan the patient to generate orthogonal images (to localize seed positions) and subsequently axially scan the patient. This sequential scanning procedure avoids using multiple independent patient scans, templates, external frames, or fiducial markers to register the reconstructed seed positions with patient contours. Dose is computed after assigning activity to (low dose rate) Ir192, linear Cs137, or I125 seeds or dwell times (high dose rate) to the Ir192 source. The planar isodose distribution is superimposed onto axial, coronal, or sagittal views of the tissues following image reconstruction. The treatment plan computes (1) direct and cumulative volume dose histograms for individual tissues, (2) the average, standard deviation, and coefficient of skewness of the dose distribution within individual tissues, (3) an average (over all tissue pixels) survival probability (S) and average survival dose DASD for a given radiation treatment, (4) normal tissue complication probability (NTCP) delivered to a given tissue. All four computed quantities account for dose heterogeneity. These estimates of the biological response to radiation from laboratory-based studies may help guide the evaluation of the prescribed low- or high-dose rate therapy in retrospective and prospective clinical studies at a number of treatment sites.


Inactivation of Snt2, a BAH/PHD-containing transcription factor, impairs pathogenicity and increases autophagosome abundance in Fusarium oxysporum.
The soil-borne, asexual fungus Fusarium oxysporum f.sp. melonis (FOM) is a causal agent of muskmelon wilt disease. The current study focused on the most virulent race of FOM-race 1,2. The tagged mutant D122, generated by Agrobacterium tumefaciens-mediated transformation, caused the delayed appearance of initial wilt disease symptoms, as well as a 75% reduction in pathogenicity. D122 was impaired in the gene product homologous to the Snt2-like transcription factor of Schizosaccharomyces pombe. Involvement of snt2 in the early stage of FOM pathogenesis and its requirement for host colonization were confirmed by targeted disruption followed by quantitative reverse transcription-polymerase chain reaction analysis of snt2 expression in planta. Δsnt2 mutants of FOM and Neurospora crassa exhibited similar morphological abnormalities, including a reduction in conidia production and biomass accumulation, slower vegetative growth and frequent hyphal septation. In N. crassa, snt-2 is required for sexual development, as Δsnt-2 mutants were unable to produce mature perithecia. Suppressive subtraction hybridization analysis of the D122 mutant versus wild-type isolate detected four genes (idi4, pdc, msf1, eEF1G) that were found previously in association with the target of rapamycin (TOR) kinase pathway. Expression of the autophagy-related idi4 and pdc genes was found to be up-regulated in the Δsnt2 FOM mutant. In N. crassa, disruption of snt-2 also conferred a significant over-expression of idi4.


Characterization of the tomato ARF gene family uncovers a multi-levels post-transcriptional regulation including alternative splicing.
The phytohormone auxin is involved in a wide range of developmental processes and auxin signaling is known to modulate the expression of target genes via two types of transcriptional regulators, namely, Aux/IAA and Auxin Response Factors (ARF). ARFs play a major role in transcriptional activation or repression through direct binding to the promoter of auxin-responsive genes. The present study aims at gaining better insight on distinctive structural and functional features among ARF proteins. Building on the most updated tomato (Solanum lycopersicon) reference genome sequence, a comprehensive set of ARF genes was identified, extending the total number of family members to 22. Upon correction of structural annotation inconsistencies, renaming the tomato ARF family members provided a consensus nomenclature for all ARF genes across plant species. In silico search predicted the presence of putative target site for small interfering RNAs within twelve Sl-ARFs while sequence analysis of the 5'-leader sequences revealed the presence of potential small uORF regulatory elements. Functional characterization carried out by transactivation assay partitioned tomato ARFs into repressors and activators of auxin-dependent gene transcription. Expression studies identified tomato ARFs potentially involved in the fruit set process. Genome-wide expression profiling using RNA-seq revealed that at least one third of the gene family members display alternative splicing mode of regulation during the flower to fruit transition. Moreover, the regulation of several tomato ARF genes by both ethylene and auxin, suggests their potential contribution to the convergence mechanism between the signaling pathways of these two hormones. All together, the data bring new insight on the complexity of the expression control of Sl-ARF genes at the transcriptional and post-transcriptional levels supporting the hypothesis that these transcriptional mediators might represent one of the main components that enable auxin to regulate a wide range of physiological processes in a highly specific and coordinated manner.


Treatment of recent-onset type 1 diabetic patients with DiaPep277: results of a double-blind, placebo-controlled, randomized phase 3 trial.
To evaluate safety and efficacy of DiaPep277 in preserving β-cell function in type 1 diabetic patients. DIA-AID 1 is a multinational, phase 3, balanced-randomized, double-blind, placebo-controlled, parallel-group clinical study. Newly diagnosed patients (N = 457, aged 16-45 years) were randomized to subcutaneous injections of DiaPep277 or placebo quarterly for 2 years. The primary efficacy end point was the change from baseline in the area under the glucagon-stimulated C-peptide curve. Secondary end points were the change from baseline in mixed-meal stimulated C-peptide secretion and in fasting C-peptide and achieving target HbA1c ≤7% (≤53 mmol/mol). Partial remission (target HbA1c on insulin ≤0.5 units/kg/day) and hypoglycemic event rate were exploratory end points. DiaPep277 was safe and well tolerated. Significant preservation of C-peptide secretion was observed in the DiaPep277-treated group compared with the placebo (relative treatment effects of 23.4%, P = 0.037, and 29.2%, P = 0.011, in the modified intent-to-treat [mITT] and per-protocol [PP] populations, respectively). The mixed-meal stimulation failed to distinguish between the groups. There was a trend toward efficacy in fasting C-peptide levels, though not statistically significant. Significantly more DiaPep277-treated than placebo-treated patients maintained target HbA1c (mITT 56% versus 44%, P = 0.03; PP 60% versus 45%, P = 0.0082) and entered partial remission (mITT 38% versus 29%, P = 0.08; PP 42% versus 30%, P = 0.035). DiaPep277 treatment reduced the relative hypoglycemic event risk (mITT by 20%; PP by 28%). DiaPep277 safely contributes to preservation of β-cell function and to improved glycemic control in patients with type 1 diabetes.


Vertebral osteoporosis in rheumatoid arthritis patients: effect of low dose prednisone therapy.
The effect of low dose prednisone therapy on spinal bone mass is controversial. We measured lumbar trabecular and cortical bone mineral density (BMD) in 74 rheumatoid arthritis (RA) patients by dual energy quantitative computerized tomography in a cross-sectional study. The presence of vertebral deformities was evaluated on a lateral spine radiograph. Patients who had never been treated with corticosteroids (n = 44) were compared with patients on long-term low dose (less than or equal to 10 mg/day) prednisone therapy (n = 30). After correction for confounding variables the lumbar BMD was highly significantly influenced by prednisone therapy in postmenopausal patients (estimated influence -31.2% on trabecular BMD and -37.2% on cortical BMD). Vertebral deformities were also significantly more frequent in prednisone treated postmenopausal patients. No negative effect of prednisone treatment could be demonstrated in male patients. In contradiction to previous reports we conclude that long-term prednisone therapy may be associated with development of spinal osteoporosis in postmenopausal RA patients, even when low doses are used.


Transvaginal sonography as a screening method for the detection of early ovarian cancer.
From December 1987 to December 1993, 6470 women underwent screening with transvaginal sonography (TVS) as part of the University of Kentucky Ovarian Cancer Screening Project. Two groups of women were eligible to participate in this investigation: (i) asymptomatic postmenopausal patients or patients >50 years of age, and (ii) asymptomatic women >30 years of age with a family history of ovarian cancer. Ovarian volume was calculated using the prolate ellipsoid formula (length x height x width x 0.523). An abnormal sonogram was defined by (1) an ovarian volume >10 cm3 in postmenopausal women or >20 cm3 in premenopausal women, and (2) a papillary or complex tissue projection into a cystic ovarian tumor. All women with an abnormal TVS had a repeat sonogram in 4-6 weeks. Patients with persistently abnormal scans had a serum CA-125 determination, tumor morphology indexing, and color Doppler sonography. Ninety patients (1.4%) with a persisting abnormality on TVS underwent exploratory laparotomy or laparoscopy for tumor removal. Thirty-seven patients had serous cystadenomas and six had primary ovarian cancers. Five patients had Stage IA ovarian cancer and one patient had Stage IIIB disease. Only one of the ovarian cancer patients had a palpable abnormality on pelvic examination, and none had an elevated (>35 u/ml) serum CA-125. All these patients are presently alive and well 1-5 years after conventional therapy. There was one false negative in this study, a 38-year-old white female who was noted to have a small ovarian cancer at the time of laparoscopic prophylactic oophorectomy 11 months after a normal scan. Over 17,000 screening years have been accrued and there have been no deaths from primary ovarian cancer in the screened population. A cost analysis of TVS screening is presented.


Immunogenicity and safety of a novel monovalent high-dose inactivated poliovirus type 2 vaccine in infants: a comparative, observer-blind, randomised, controlled trial.
Following the proposed worldwide switch from trivalent oral poliovirus vaccine (tOPV) to bivalent types 1 and 3 OPV (bOPV) in 2016, inactivated poliovirus vaccine (IPV) will be the only source of protection against poliovirus type 2. With most countries opting for one dose of IPV in routine immunisation schedules during this transition because of cost and manufacturing constraints, optimisation of protection against all poliovirus types will be a priority of the global eradication programme. We assessed the immunogenicity and safety of a novel monovalent high-dose inactivated poliovirus type 2 vaccine (mIPV2HD) in infants. This observer-blind, comparative, randomised controlled trial was done in a single centre in Panama. We enrolled healthy infants who had not received any previous vaccination against poliovirus. Infants were randomly assigned (1:1) by computer-generated randomisation sequence to receive a single dose of either mIPV2HD or standard trivalent IPV given concurrently with a third dose of bOPV at 14 weeks of age. At 18 weeks, all infants were challenged with one dose of monovalent type 2 OPV (mOPV2). Primary endpoints were seroconversion and median antibody titres to type 2 poliovirus 4 weeks after vaccination with mIPV2HD or IPV; and safety (as determined by the proportion and nature of serious adverse events and important medical events for 8 weeks after vaccination). The primary immunogenicity analyses included all participants for whom a post-vaccination blood sample was available. All randomised participants were included in the safety analyses. This trial is registered with ClinicalTrials.gov, number NCT02111135. Between April 14 and May 9, 2014, 233 children were enrolled and randomly assigned to receive mIPV2HD (117 infants) or IPV (116 infants). 4 weeks after vaccination with mIPV2HD or IPV, seroconversion to poliovirus type 2 was recorded in 107 (93·0%, 95% CI 86·8-96·9) of 115 infants in the mIPV2HD group compared with 86 (74·8%, 65·8-82·4) of 115 infants in the IPV group (difference between groups 18·3%, 95% CI 5·0-31·1; p<0·0001), and median antibody titres against poliovirus type 2 were 181 (95% CI 72·0-362·0) in the mIPV2HD group and 36 (18·0-113·8) in the IPV group (difference between groups 98·8, 95% CI 60·7-136·9; p<0·0001). Serious adverse events were reported for six (5%) of 117 infants in the mIPV2HD group and seven (6%) of 116 infants in the IPV group during the 8-week period after vaccination; none were related to vaccination. No important medical events were reported. Our findings lend support to the use of mIPV2HD as an option for stockpiling for outbreak response or primary protection in selected areas at risk for emergence of poliovirus type 2 during the next phase of the polio eradication plan. Bill & Melinda Gates Foundation.


Relationship between dietary factors and prostate-specific antigen in healthy men.
There is little evidence indicating whether dietary factors influence prostate-specific antigen (PSA) concentrations. We examined whether nutritional factors, including energy, protein, fat, and carbohydrate intake were associated with PSA in healthy men. We investigated 13,594 men aged 50 years and over who visited a hospital for a routine health checkup between 2003 and 2007. Dietary intake was assessed using a food frequency questionnaire. We performed a multiple linear regression to examine the association between PSA and dietary intake. After controlling for age, body mass index, and physical activity, PSA was significantly negatively associated with percent protein intake (p for trend < 0.001). Compared with the lowest quintile, PSA was 5.8% lower (95% CI: -8.9 to -2.5%) in the highest quintile. We also observed a significant positive association between percent fat intake and PSA concentration (p for trend 0.043). PSA was 3.4% greater (95% CI: 0-6.9%) among men in the highest quintile compared with those in the lowest quintile. Men who had a lower percent protein intake and higher percent fat intake had an elevated PSA level, although the magnitude of these associations was small.


Clearing the cervical spine in the blunt trauma patient.
The goal of cervical spine clearance is to establish that injuries are not present. Patients are classified into four groups: asymptomatic, temporarily nonassessable secondary to distracting injuries or intoxication, symptomatic, and obtunded. Level I evidence supports that the asymptomatic patient can be cleared on clinical grounds and does not require imaging. The temporarily nonassessable patient may have short-term mental status changes (eg, intoxication, painful distracting injuries) and can be evaluated by two methods. When there is urgency, the evaluation is similar to that for the obtunded patient. Alternatively, the patient can be reevaluated within 24 to 48 hours, after return of mentation or following treatment of painful injuries. The patient then can be assessed as the asymptomatic patient is. The symptomatic patient requires advanced imaging. The obtunded patient should undergo, at minimum, a multidetector CT scan. Two methods are advocated. One uses only multidetector CT; a normal result is sufficient to clear the obtunded patient. The alternative method is obtaining a magnetic resonance image subsequent to a negative multidetector CT scan. Because at present information is insufficient to determine whether MRI is indicated, this is an area of controversy.


[Characteristics of the changes in mental activity of children with pathology of the biliary tract].
The paper treats of the data on the psychoneurological sphere in preschool children suffering from dyskinesia of the biliary tract. A study was made of the psychoneurological status of 120 schoolchildren with biliary pathology. Three variants of the +cerebro-asthenic syndrome were revealed: ++astheno-adynamic, ++astheno-hyperdynamic and moderately marked ones. To objectively evaluate the mental status and the structure of mental activity disorders, all the children underwent an experimental and psychological examination. A relationship was discovered between the changes in mental activity and the variant of the cerebrasthenic syndrome and the child's age. The most remarkable changes in mental activity were recorded in children with the ++astheno-adynamic variant of the +cerebro-asthenic syndrome.


[Associations between the renal tubular acidosis of primary Sjogren syndrome and the infection of Epstein-Barr virus: a preliminary study].
To explore the possible etiological associations between the infection of Epstein-Barr Virus (EBV) and the occurrence of renal tubular acidosis (RTA) of primary Sjogren's syndrome (SS), renal biopsies from both primary SS with clinical or subclinical RTA and controls were detected by using a monoclonal antibody against EBV early antigen P138 and a 32P-labelled EBV Bam W probe. As a result, cytoplasmic fluorescence staining of epithelial cells with the monoclonal antibody as described above was noted in all the seven renal biopsies from primary SS, and one out of two renal samples among them contained EBV DNA detectable by dot blot hybridization with Bam W probe. Whereas five control renal biopsies were all negative on the same methods of detection. Our study suggested that EBV has been of a lytic fate leading to active replication in the kidneys of patients with RTA of primary SS. Therefore, EBV may play an important role in the renal damage, especially resulting in RTA, of primary SS.


Comparative study of membrane potential-sensitive fluorescent probes and their use in ion channel screening assays.
In this study, the authors compared and evaluated 4 membrane potential probes in the same cellular assay: the oxonol dye DiBAC(4)(3), the FLIPR membrane potential (FMP) dye (Molecular Devices), and 2 novel fluorescence resonance energy transfer (FRET) dye systems from PanVera [CC2-DMPE/DiSBAC(2)(3)] and Axiom [DiSBAC(1)(3)/DiSBAC(1)(5)]. The kinetic parameters of each membrane probe were investigated in RBL-2H3 cells expressing an endogenous inward rectifier potassium channel (IRK1). The FMP dye presented the highest signal over background ratio whereas the FRET dyes from PanVera gave the fastest response. The determination of IC(50) values for 8 different channel modulators indicated a good correlation between the 4 membrane probe systems. The compound-dye interaction was evaluated in the presence of compounds at 10 muM and clearly indicated no effect on the FMP or the PanVera donor dye, whereas some major interference with the oxonol probes was observed. Using a cell permeabilization assay in the presence of gramicidin, the authors concluded that the FRET dyes from PanVera and the FMP dye are unable to measure the gramicidin-induced cell membrane hyperpolarizations. The 4 dye systems were investigated under high-throughput screening (HTS) conditions, and their respective Z' parameter was determined. The characteristics of each dye system and its potential use in HTS assays is discussed.


Alpha1,3-galactosyltransferase gene-knockout pigs for xenotransplantation: where do we go from here?
The ability to genetically engineer pigs that no longer express the Galalpha1,3Gal (Gal) oligosaccharide has been a significant step toward the clinical applicability of xenotransplantation. Using a chronic immunosuppressive regimen based on costimulatory blockade, hearts from these pigs have survived from 2 to 6 months in baboons. Graft failure was predominantly from the development of a thrombotic microangiopathy. Potential contributing factors include the presence of preformed anti-nonGal antibodies or the development of low levels of elicited antibodies to nonGal antigens, natural killer (NK) cell or macrophage activity, and inherent coagulation dysregulation between pigs and primates. The breeding of pigs transgenic for an "anticoagulant" gene, such as human tissue factor pathway inhibitor, hirudin, or CD39, or lacking the gene for the prothrombinase, fibrinogen-like protein-2, is anticipated to inhibit the change in the endothelium to a procoagulant state that takes place in the pig organ after transplantation. The identification of the targets for anti-nonGal antibodies and/or human macrophages might allow further genetic modification of the pig, and xenogeneic NK cell recognition and activation may be inhibited by the transgenic expression of human leukocyte antigen molecules and/or by blocking the function of activating NK receptors. The ultimate goal of induction of T-cell tolerance may be possible only if these hurdles in the coagulation system and innate immunity can be overcome.


Long-run Relations between Childhood Shocks and Health in Late Adulthood-Evidence from the Survey of Health, Ageing, and Retirement in Europe.
In this article, we address the long-run associations between childhood shocks and health in late adulthood. Applying a life-course approach and data from SHARE, we estimate direct and indirect relations of shocks like relocation, dispossession, or hunger and health outcomes after 50 years of age. Having lived in a children's home, in a foster family, or having suffered a period of hunger turn out to be the most detrimental. Using a finite mixture model, which allows to classify the correlations between shocks and later health into a priori unknown groups, we show that some adverse shocks show opposite relations for specific groups. (JEL codes: J1, I12, J13).


Keratouveitis from Euphorbia cyparissias exposure is a temporal phenomenon.
We present the case of an 82-year-old woman with severe keratouveitis secondary to exposure to the plant Euphorbia cyparissias during the month of August. Despite copious irrigation of both eyes, she developed a case of Euphorbia keratouveitis that was typical in its course. Exposure to plant sap from this species should be treated much like an alkaline chemical injury as the pH of the ocular surface may be elevated. E. cyparissias should be carefully handled as it may result in keratouveitis, especially in early autumn.


The association between grip strength and carotid intima media thickness: A Mendelian randomization analysis of the Canadian Longitudinal Study on Aging.
Several two-sample Mendelian randomization studies have reported discordant results concerning the association between grip strength and cardiovascular disease, possibly due to the number of instrumental variables used, pleiotropic bias, and/ or effect modification by age and sex. We conducted a sex- and age-stratified one-sample Mendelian randomization study in the Canadian Longitudinal Study on Aging. We investigated whether grip strength is associated with carotid intima media thickness (cIMT), a marker of vascular atherosclerosis event risk, using eighteen single nucleotide polymorphisms (SNP) identified as specifically associated with grip strength. A total of 20,258 participants of self-reported European ancestry were included in the analytic sample. Our Mendelian randomization findings suggest a statistically significant association between grip strength and cIMT (MR coefficient of 0.02 (95% CI: 0.01, 0.04)). We found no statistically significant differences between sexes (p-value = 0.201), or age groups [(≤ 60 years old versus >60 years old); p-value = 0.421]. This study provides evidence that grip strength is inversely associated with cIMT. Our one-sample MR study design allowed us to demonstrate that there is no evidence of heterogeneity of effects according to age group or biological sex.


Insect wing deformation measurements using high speed digital holographic interferometry.
An out-of-plane digital holographic interferometry system is used to detect and measure insect's wing micro deformations. The in-vivo phenomenon of the flapping is registered using a high power cw laser and a high speed camera. A series of digital holograms with the deformation encoded are obtained. Full field deformation maps are presented for an eastern tiger swallowtail butterfly (Pterourus multicaudata). Results show no uniform or symmetrical deformations between wings. These deformations are in the order of hundreds of nanometers over the entire surface. Out-of-plane deformation maps are presented using the unwrapped phase maps.


S-100 immunoreactivity in rat brain glial cultures is associated with both astrocytes and oligodendrocytes.
S-100 protein, a Ca(2+)-binding protein of the EF-hand type, is most abundant in the brain, and is involved in cell differentiation and the molecular mechanisms underlying cytoskeletal organization. We have investigated the immunocytochemical localization of S-100 protein in rat brain glial cultures prepared from the cerebral hemispheres of newborn rats. In mixed glial cultures, containing astrocytes type I and II and oligodendrocytes at various stages of differentiation, S-100 immunoreactivity was detected in all three cell types. Double immunofluorescence analysis revealed that in astrocytes, S-100 immunoreactivity was mainly colocalized with glial fibrillary acidic protein (GFAP), while in oligodendrocytes a close association with microtubular structures was observed. For immunoblot analysis, highly enriched oligodendrocytes and astrocytes were separately cultured for another week, and their extracts were analyzed by immunoblotting. The immunoblots of the cell extracts of both cell types showed a single S-100-immunoreactive polypeptide with an apparent molecular weight of approximately 12,000 daltons. Thus, the data presented here demonstrate that S-100 protein is not confined to astrocytes but occurs also in oligodendrocytes of rat brain. The close association with the oligodendroglial cytoskeleton suggests that this protein could also play a regulatory role in the organization of microtubules in oligodendrocytes and hence may be involved in the formation and maintenance of the myelin-containing membrane sheets.


The Role of Bcl-2 and Beclin-1 Complex in "Switching" between Apoptosis and Autophagy in Human Glioma Cells upon LY294002 and Sorafenib Treatment.
Gliomas are the most malignant tumors of the central nervous system. One of the factors in their high drug resistance is avoiding programmed death (PCD) induction. This is related to the overexpression of intracellular survival pathways: PI3K-Akt/PKB-mTOR and Ras-Raf-MEK-ERK. Apoptosis and autophagy are co-existing processes due to the interactions between Bcl-2 and beclin-1 proteins. Their complex may be a molecular "toggle-switch" between PCD types. The aim of this research was to investigate the role of Bcl-2:beclin-1 complex in glioma cell elimination through the combined action of LY294002 and sorafenib. Drug cytotoxicity was estimated with an MTT test. The type of cell death was evaluated using variant microscopy techniques (fluorochrome staining, immunocytochemistry, and transmission electron microscopy), as well as the Bcl-2:beclin-1 complex formation and protein localization. Molecular analysis of PCD indicators was conducted through immunoblotting, immunoprecipitation, and ELISA testing. SiRNA was used to block Bcl-2 and beclin-1 expression. The results showed the inhibitors used in simultaneous application resulted in Bcl-2:beclin-1 complex formation and apoptosis becoming dominant. This was accompanied by changes in the location of the tested proteins. "Switching" between apoptosis and autophagy using PI3K and Raf inhibitors with Bcl-2:beclin-1 complex formation opens new therapeutic perspectives against gliomas.


Glucose administration and cognitive function: differential effects of age and effort during a dual task paradigm in younger and older adults.
Current research suggests that glucose facilitates performance on cognitive tasks which possess an episodic memory component and a relatively high level of cognitive demand. However, the extent to which this glucose facilitation effect is uniform across the lifespan is uncertain. This study was a repeated measures, randomised, placebo-controlled, cross-over trial designed to assess the cognitive effects of glucose in younger and older adults under single and dual task conditions. Participants were 24 healthy younger (average age 20.6 years) and 24 healthy older adults (average age 72.5 years). They completed a recognition memory task after consuming drinks containing 25 g glucose and a placebo drink, both in the presence and absence of a secondary tracking task. Glucose enhanced recognition memory response time and tracking precision during the secondary task, in older adults only. These findings do not support preferential targeting of hippocampal function by glucose, rather they suggest that glucose administration differentially increases the availability of attentional resources in older individuals.


Depression and intelligence in patients with Parkinson's disease and deep-brain stimulation.
The goal of this study is to examine the association of depression with intelligence and education in patients with Parkinson's disease treated with bilateral subthalamic nucleus stimulation (STN-DBS). The literature has been contradictory concerning depression in Parkinson's disease patients. Some studies have shown less depression in Parkinson's disease patients with more education not treated with STN-DBS. Other recently published studies indicate that STN-DBS improves the depression associated with Parkinson's disease. No studies have examined the correlation of these factors with depression in Parkinson's disease patients treated with STN-DBS. We administered the Beck Depression Inventory (BDI) pre- and postoperatively to 21 Parkinson's disease patients (seven women, 14 men, ages 49-75) who underwent STN-DBS. The postoperative scores of the lower 50th percentile (n=8) of the Verbal Comprehensive Index of the Wechsler Adult Intelligence Scale (WAIS-III) decreased significantly (P=0.036), while the upper 50th percentile (n=13) remained nearly constant (P=0.802). Furthermore, as the education increased from highschool to graduate level, patients demonstrated less improvement in depressive symptoms postoperatively. These findings suggest that Parkinson's disease patients with lower intelligence test scores and less education benefit more with regards to depressive symptomatology after STN-DBS than patients with higher scores and education.


Synergism versus Additivity: Defining the Interactions between Common Disinfectants.
Many of the most common disinfectant and sanitizer products are formulations of multiple antimicrobial compounds. Products claiming to contain synergistic formulations are common, although there is often little supporting evidence. The antimicrobial interactions of all pairwise combinations of common disinfectants (benzalkonium chloride, didecyldimethylammonium chloride, polyhexamethylene biguanide, chlorocresol, and bronopol) were classified via checkerboard assay and validated by time-kill analyses. Combinations were tested against Acinetobacter baumannii NCTC 12156, Enterococcus faecalis NCTC 13379, Klebsiella pneumoniae NCTC 13443, and Staphylococcus aureus NCTC 13143. Synergistic interactions were identified only for the combinations of chlorocresol with benzalkonium chloride and chlorocresol with polyhexamethylene biguanide. Synergism was not ubiquitously demonstrated against all species tested and was on the borderline of the synergism threshold. These data demonstrate that synergism between disinfectants is uncommon and circumstantial. Most of the antimicrobial interactions tested were characterized as additive. We suggest that this is due to the broad, nonspecific mechanisms associated with disinfectants not providing an opportunity for the combined activities of these compounds to exceed the sum of their parts. IMPORTANCE The scarcity of observed synergistic interactions suggests that in the case of many disinfectant-based products, combined mechanisms of interaction may be being misinterpreted. We emphasize the need to correctly differentiate between additivity and synergism in antimicrobial formulations, as inappropriate classification may lead to unnecessary issues in the event of regulatory changes. Furthermore, we question the need to focus on synergism and disregard additivity when considering combinations of disinfectants, as the benefits that synergistic interactions provide are not necessarily relevant to the application of the final product.


STRA13 interacts with STAT3 and modulates transcription of STAT3-dependent targets.
STRA13 is a pVHL-dependent bHLH transcription factor up-regulated on the mRNA level in multiple cancer cell lines and implicated recently in the regulation of immune cell homeostasis and autoimmunity. In searching for STRA13-interacting proteins with oncogenic potential by the yeast two-hybrid screening, we identified STAT3 beta as a STRA13-binding partner. We showed that STRA13 binds predominantly to phosphorylated (active) STAT3 alpha and beta isoforms via its HLH and C-terminal regions. We also found that STRA13 was able to activate transcription from STAT-dependent cis-elements. Expression of endogenous STRA13 was shown to be cytokine-inducible, consistent with STRA13 involvement in STAT-dependent transcription regulation. We demonstrated that the STAT3-regulated promoter of the pro-apoptotic Fas gene was activated upon STRA13 over-expression and that co-expression of STRA13 with STAT3 beta or STAT3 alpha modulated the transcriptional outcome. Forced expression of STRA13 induced apoptosis, in agreement with the STRA13 activation effect on the Fas promoter. Simultaneous expression of STRA13 and STAT3 beta resulted in alleviation of the STRA13 pro-apoptotic effect. Thus, for the first time, we identify STRA13 as a STAT3 partner and provide a consistent line of evidence for STRA13 involvement into regulation of apoptosis via the STAT pathways.


Effects of carbon and oxygen limitations and calcium concentrations on biofilm removal processes.
Bacterial biofilm removal processes due to shear and catastrophic sloughing have been investigated in a turbulent flow system under conditions of carbon versus oxygen substrate limitations and varying aqueous phase calcium concentrations. Biofilm cellular and extracellular polymer carbon, total biofilm carbon and mass, and biofilm calcium concentrations are measured for pure culture biofilms of the facultative aerobe, Pseudomonas putida ATCC 11172. Results indicate oxygen-limited biofilms reach a higher steady-state biofilm organic carbon level than carbon-limited biofilms. Oxygen-limited biofilms also exhibit (1) a higher extracellular polymer-carbon: cell-carbon ratio throughout biofilm development and (2) a higher biofilm calcium content than carbon-limited biofilms. Increasing aqueous phase calcium concentrations increase the amount of biofilm calcium in both cases; the rate of calcium accumulation in oxygen-limited biofilms increases with increasing liquid phase calcium concentrations over the entire range studied while the rates of calcium accumulation in carbon-limited biofilms appear independent of aqueous phase calcium concentrations above 11.0 mg/L. Oxygen-limited biofilms with their higher extracellular polymer and calcium content exhibit shear removal rates that are 20-40% of those observed for carbon-limited biofilms. However, it is the oxygen-limited biofilms that experience catastrophic sloughing events. The carbon-limited biofilms studied here never sloughed even if subjected to intentional long-term deprivation of all nutrients. Reduced shear removal and the susceptibility to sloughing of the oxygen-limited biofilms are attributed to their more cohesive structure bought about by their relatively greater extracellular polymer production.


Relevance of the chronobiological and non-chronobiological actions of melatonin for enhancing therapeutic efficacy in neurodegenerative disorders.
Melatonin is an indolamine with a large spectrum of functions that can be divided into chronobiotic and nonchronobiotic. Chronobiotic effects are mediated by the daily rhythm of melatonin in the plasma due to nocturnal pineal synthesis, whereas the melatonin produced by other cells, such as gastrointestinal and immune competent cells, is independent of the light/dark cycle and exert non-chronobiotic effects. The concentrations achieved by the two sources are significantly different, varying in the pM-nM range in the plasma, and may achieve concentrations in the mM range when released locally by activated immune-competent cells. Consequently, the effects of the melatonin produced in these two situations are distinct. Much has been reported about the beneficial response to exogenous melatonin administration in several pathological conditions. However, the relationship between the establishment of a disease and the state of the physiological activity of the pineal gland is still poorly understood. Here, we review the state of art in the modulation of pineal melatonin synthesis, relevant patents, and discuss its relationship with neurodegenerative disorders that involve a central inflammatory response, such as Alzheimer's disease, to suggest the putative relevance of new therapeutic protocols that replace this pineal hormone.


From Mesenchymal Stromal Cells to Engineered Extracellular Vesicles: A New Therapeutic Paradigm.
Mesenchymal stromal cells (MSCs) are multipotent cells obtained from many tissues including bone marrow, adipose tissue, umbilical cord, amniotic fluid, and placenta. MSCs are the leading cell source for stem cell therapy due to their regenerative and immunomodulatory properties, their low risk of tumorigenesis and lack of ethical constraints. However, clinical applications of MSCs remain limited. MSC therapeutic development continues to pose challenges in terms of preparation, purity, consistency, efficiency, reproducibility, processing time and scalability. Additionally, there are issues with their poor engraftment and survival in sites of disease or damage that limit their capacity to directly replace damaged cells. A key recent development in MSC research, however, is the now widely accepted view that MSCs primarily exert therapeutic effects via paracrine factor secretion. One of the major paracrine effectors are extracellular vesicles (EVs). EVs represent a potential cell-free alternative to stem cell therapy but are also rapidly emerging as a novel therapeutic platform in their own right, particularly in the form of engineered EVs (EEVs) tailored to target a broad range of clinical indications. However, the development of EVs and EEVs for therapeutic application still faces a number of hurdles, including the establishment of a consistent, scalable cell source, and the development of robust GMP-compliant upstream and downstream manufacturing processes. In this review we will highlight the clinical challenges of MSC therapeutic development and discuss how EVs and EEVs can overcome the challenges faced in the clinical application of MSCs.


Selective adsorption of L-serine functional groups on the anatase TiO2(101) surface in benthic microbial fuel cells.
To help design bacteria-friendly anodes for unmediated benthic microbial fuel cells (MFCs), we explore the role of anatase TiO2(101) surface biocompatibility in selecting the functional groups of the levo-isomer serine (L-Ser), which contains carboxyl, hydroxyl, and amino groups in a single molecule. By performing total energy calculations and molecular dynamics simulations based on a density-functional tight-binding method, we find that at room temperature, the surface should be active for biomolecules with carboxyl/carboxylic and hydroxyl groups, but it is not sensitive to those with amino groups. The hydrogen bonding between the hydroxyl H and surface O facilitates electron transfer from the pili or the bacterial matrix to the anode surface, which improves the output power density. Thus, in combination with conductive polymers, the anatase TiO2(101) surface can be an effective biocompatible substrate in benthic MFCs by enabling the surface O to form more hydrogen bonds with the hydroxyl H of the biomolecule.


The chiral potential of phenanthriplatin and its influence on guanine binding.
The monofunctional platinum complex cis-[Pt(NH3)2Cl(Am)](+), also known as phenanthriplatin, where Am is the N-heterocyclic base phenanthridine, has promising anticancer activity. Unlike bifunctional compounds such as cisplatin, phenanthriplatin can form only one covalent bond to DNA. Another distinguishing feature is that phenanthriplatin is chiral. Rotation about the Pt-N bond of the phenanthridine ligand racemizes the complex, and the question arises as to whether this process is sufficiently slow under physiological conditions to impact its DNA-binding properties. Here we present the results of NMR spectroscopic, X-ray crystallographic, molecular dynamics, and density functional theoretical investigations of diastereomeric phenanthriplatin analogs in order to probe the internal dynamics of phenanthriplatin. These results reveal that phenanthriplatin rapidly racemizes under physiological conditions. The information also facilitated the interpretation of the NMR spectra of small molecule models of phenanthriplatin-platinated DNA. These studies indicate, inter alia, that one diastereomeric form of the complexes cis-[Pt(NH3)2(Am)(R-Gua)](2+), where R-Gua is 9-methyl- or 9-ethylguanine, is preferred over the other, the origin of which stems from an intramolecular interaction between the carbonyl oxygen of the platinated guanine base and a cis-coordinated ammine. The relevance of this finding to the DNA-damaging properties of phenanthriplatin and its biological activity is discussed.


Three-dimensional methods in seismic exploration.
Two-dimensional seismic prospecting methods are inadequate to solve the three-dimensional geological problems that arise in petroleum exploration. The three-dimensional seismic technique is a data collection and processing system that samples the subsurface adequately. The structure and lithology of rocks are mapped accurately by imaging their acoustic reflectivity with the scalar wave equation. In oil industry economics, the expense of three-dimensional seismic methods is justified by their usefulness for discovering additional reserves and for reducing the growing costs of developing oil and gas fields.


A generalized model for predicting postcompletion errors.
A postcompletion error is a type of procedural error that occurs after the main goal of a task has been accomplished. There is a strong theoretical foundation accounting for postcompletion errors (Altmann & Trafton, 2002; Byrne & Bovair, 1997). This theoretical foundation has been leveraged to develop a logistic regression model of postcompletion errors based on reaction time and eye movement measures (Ratwani, McCurry, & Trafton, 2008). This study further develops and extends this predictive model by (a) validating the model and the general set of predictors on a new task to test the robustness of the model, and (b) determining which specific theoretical components are most important to postcompletion error prediction.


FPGA Realizations of Chaotic Epidemic and Disease Models Including Covid-19.
The spread of epidemics and diseases is known to exhibit chaotic dynamics; a fact confirmed by many developed mathematical models. However, to the best of our knowledge, no attempt to realize any of these chaotic models in analog or digital electronic form has been reported in the literature. In this work, we report on the efficient FPGA implementations of three different virus spreading models and one disease progress model. In particular, the Ebola, Influenza, and COVID-19 virus spreading models in addition to a Cancer disease progress model are first numerically analyzed for parameter sensitivity via bifurcation diagrams. Subsequently and despite the large number of parameters and large number of multiplication (or division) operations, these models are efficiently implemented on FPGA platforms using fixed-point architectures. Detailed FPGA design process, hardware architecture and timing analysis are provided for three of the studied models (Ebola, Influenza, and Cancer) on an Altera Cyclone IV EP4CE115F29C7 FPGA chip. All models are also implemented on a high performance Xilinx Artix-7 XC7A100TCSG324 FPGA for comparison of the needed hardware resources. Experimental results showing real-time control of the chaotic dynamics are presented.