Calcium kinetics during bed rest with artificial gravity and exercise countermeasures.
We assessed the potential for countermeasures to lessen the loss of bone calcium during bed rest. Subjects ingested less calcium during bed rest, and with artificial gravity, they also absorbed less calcium. With exercise, they excreted less calcium. To retain bone during bed rest, calcium intake needs to be maintained. This study aims to assess the potential for artificial gravity (AG) and exercise (EX) to mitigate loss of bone calcium during space flight. We performed two studies: (1) a 21-day bed rest (BR) study with subjects receiving 1 h/day AG (n = 8) or no AG (n = 7) and (2) a 28-day BR study with 1 h/day resistance EX (n = 10) or no EX (n = 3). In both studies, stable isotopes of Ca were administered orally and intravenously, at baseline and after 10 days of BR, and blood, urine, and feces were sampled for up to 14 days post dosing. Tracers were measured using thermal ionization mass spectrometry. Data were analyzed by compartmental modeling. Less Ca was absorbed during BR, resulting in lower Ca balance in BR+AG (-6.04 ± 3.38 mmol/day, P = 0.023). However, Ca balance did not change with BR+EX, even though absorbed Ca decreased and urinary Ca excretion increased, because endogenous excretion decreased, and there was a trend for increased bone deposition (P = 0.06). Urinary N-telopeptide excretion increased in controls during BR, but not in the EX group. Markers of bone formation were not different between treatment groups for either study. Ca intake decreased during BR (by 5.4 mmol/day in the AG study and 2.8 mmol/day in the EX study), resulting in lower absorbed Ca. During BR (or space flight), Ca intake needs to be maintained or even increased with countermeasures such as exercise, to enable maintenance of bone Ca.


Do small airway abnormalities characterize asthma phenotypes? In search of proof.
The role of small airway abnormalities in asthma pathogenesis has been extensively studied and debated for several decades. However, whether or not small airway abnormalities play a relevant role in specific phenotypes of asthmatic patients and contribute to clinical presentation is largely unknown. In the present review, we evaluated available data on the role of small airways in severe asthma, with a further focus on asthma in smokers and asthma in the elderly. These phenotypes are characterized by a poor response to treatment and they can represent a model of greater small airway impairment. In severe asthmatics, small airway involvement has been shown through evidence of both distal inflammation and of increased air trapping. The few available data on asthmatics who smoke, and elderly asthmatics, similarly suggests that small airway abnormalities contribute to the pathogenesis of the disease. In this perspective, there could be a rationale for specifically assessing small airway impairment in these patients and for clinical studies evaluating whether pharmacological approaches targeting the more peripheral airways result in clinical benefits beyond conventional therapy.


Morphologically controlled synthesis of colloidal upconversion nanophosphors and their shape-directed self-assembly.
We report a one-pot chemical approach for the synthesis of highly monodisperse colloidal nanophosphors displaying bright upconversion luminescence under 980 nm excitation. This general method optimizes the synthesis with initial heating rates up to 100 °C/minute generating a rich family of nanoscale building blocks with distinct morphologies (spheres, rods, hexagonal prisms, and plates) and upconversion emission tunable through the choice of rare earth dopants. Furthermore, we employ an interfacial assembly strategy to organize these nanocrystals (NCs) into superlattices over multiple length scales facilitating the NC characterization and enabling systematic studies of shape-directed assembly. The global and local ordering of these superstructures is programmed by the precise engineering of individual NC's size and shape. This dramatically improved nanophosphor synthesis together with insights from shape-directed assembly will advance the investigation of an array of emerging biological and energy-related nanophosphor applications.


Evidence for the presence of two conformations of the heme a3-Cu(B) pocket of cytochrome caa3 from Thermus thermophilus.
Resonance Raman (RR) and "light" minus "dark" Fourier transform infrared (FTIR) difference spectra are reported for the CO-bound caa(3) oxidase from Thermus thermophilus. Two Fe-CO stretching modes at 518 and 507 cm(-1), the Fe-C-O bending mode at 570 cm(-1), and three C-O modes of heme a(3) at 1958, 1967, and 1973 cm(-1) have been identified in the RR and FTIR spectra, respectively. The FTIR "light" minus "dark" spectrum indicates the formation of Cu(B)CO as revealed by its ν(CO) at 2060/2065 cm(-1). We assign the bands at 518 (ν(Fe-CO)) and 1967/1973 cm(-1) (ν(C-O)) as the α-conformation. We also assign the bands at 507 and 1958 cm(-1) (ν(C-O)) as originating from the β-conformation of the enzyme. A frequency upshift of the heme a(3) Fe-His mode is observed subsequent to CO photolysis from 209 cm(-1) in the equilibrium deoxy enzyme to 214 cm(-1) in the photoproduct. The caa(3) data, distinctly different from those of ba(3) oxidase, are discussed in terms of the coupling of the α- and β-conformations that occur in heme-copper oxidases with catalytic function. The dynamics between the heme a(3) and heme a propionates as revealed by the perturbation of the bending vibrations δ(prop) of hemes a and a(3) at 385 and 392 cm(-1), respectively, induced upon CO binding to heme a(3) is discussed in terms of the protonic connectivity between the heme a ring-D propionate/Arg site with that of the heme a(3) ring-D propionate-H(2)O site that leads to the highly conserved in the heme-copper oxidases water pool.


Quantifying the Impact of Cancer on the Viscoelastic Properties of the Prostate Gland using a Quasi-Linear Viscoelastic Model.
Pathological disorders can alter the mechanical properties of biological tissues, and studying such changes can help to better understand the disease progression. The prostate gland is no exception, as previous studies have shown that cancer can affect its mechanical properties. However, most of these studies have focused on the elastic properties of the tissue and have overlooked the impact of cancer on its viscous response. To address this gap, we used a quasi-linear viscoelastic model to investigate the impact of cancer on both the elastic and viscous characteristics of the prostate gland. By comparing the viscoelastic properties of segments influenced by cancer and those unaffected by cancer in 49 fresh prostates, removed within two hours after prostatectomy surgery, we were able to determine the influence of cancer grade and tumor volume on the tissue. Our findings suggest that tumor volume significantly affects both the elastic modulus and viscosity of the prostate (p-value less than 2%). Specifically, we showed that cancer increases Young's modulus and shear relaxation modulus by 20%. These results have implications for using mechanical properties of the prostate as a potential biomarker for cancer. However, developing an in vivo apparatus to measure these properties remains a challenge that needs to be addressed in future research. STATEMENT OF SIGNIFICANCE: This study is the first to explore how cancer impacts the mechanical properties of prostate tissues using a quasi-linear viscoelastic model. We examined 49 fresh prostate samples collected immediately after surgery and correlated their properties with cancer presence identified in pathology reports. Our results demonstrate a 20% change in the viscoelastic properties of the prostate due to cancer. We initially validated our approach using tissue-mimicking phantoms and then applied it to differentiate between cancerous and normal prostate tissues. These findings offer potential for early cancer detection by assessing these properties. However, conducting these tests in vivo remains a challenge for future research.


Advances in technology for management of type 1 diabetes.
Technological advances have had a major effect on the management of type 1 diabetes. In addition to blood glucose meters, devices used by people with type 1 diabetes include insulin pumps, continuous glucose monitors, and, most recently, systems that combine both a pump and a monitor for algorithm-driven automation of insulin delivery. In the next 5 years, as many advances are expected in technology for the management of diabetes as there have been in the past 5 years, with improvements in continuous glucose monitoring and more available choices of systems that automate insulin delivery. Expansion of the use of technology will be needed beyond endocrinology practices to primary-care settings and broader populations of patients. Tools to support decision making will also need to be developed to help patients and health-care providers to use the output of these devices to optimise diabetes management.


The Substituent Effect on the Radical Scavenging Activity of Apigenin.
Flavonoids widely found in natural foods are excellent free radical scavengers. The relationship between the substituent and antioxidative activity of flavonoids has not yet been completely elucidated. In this work, the antioxidative activity of apigenin derivatives with different substituents at the C3 position was determined by density functional theory (DFT) calculations. The bond dissociation enthalpy (BDE), ionization potential (IP), and proton affinity (PA) were calculated. Donator acceptor map (DAM) analysis illustrated that the studied compounds are worse electron acceptors than F and also are not better electron donors than Na. The strongest antioxidative group of apigenin derivatives was the same as apigenin. Excellent correlations were found between the BDE/IP/PA and Hammett sigma constants. Therefore, Hammett sigma constants can be used to predict the antioxidative activity of substituted apigenin and to design new antioxidants based on flavonoids. In non-polar phases, the antioxidative activity of apigenin was increased by the electron-withdrawing groups, while it was reduced by the electron-donating groups. Contrary results occurred in the polar phase. The electronic effect of the substituents on BDE(4'-OH), BDE(5-OH), PA(4'-OH), and IP is mainly controlled by the resonance effect, while that on BDE(7-OH), PA(5-OH), and PA(7-OH) is governed by the field/inductive effect.


A dangerous freshwater contact.
An 8-year-old girl was presented to our department for persistent diarrhoea. A first diagnosis of tuberculosis, along with the result of the chest x-ray scan, had been posed some months before, after a holiday in Brazil, when she started presenting aspecific systemic and gastrointestinal symptoms. The girl was under specific antitubercular treatment when we first saw her. New diagnosis of schistosomiasis was posed by our laboratory tests. Treatment with praziquantel was started and complete resolution of clinical and radiological picture was observed.


Lymphatic and Blood Network Analysis During Obesity.
Lymphatic collecting vessels and lymph nodes are inevitably embedded in adipose tissue. The physiological significance of this observation remains still not elucidated. However, obesity is characterized by impaired lymphatic function and increased vessel permeability. Inversely, lymphatic dysfunction induces obesity in mice, suggesting a significant interplay between lymphatic vessels and the adipose tissue. Therefore, understanding factors leading to lymphatic dysfunction might open new therapeutic windows to prevent obesity and associated comorbidities. The first step in this process requires a precise and detailed visualization of the lymphatic network in healthy and inflamed adipose tissue. Here, we describe a rapid, inexpensive, and efficient method that allows to label and analyze lymphatic and blood vessels. This approach takes advantage of the skin-draining brachial lymph node localization within the subcutaneous adipose tissue. The lymphatic arborization of this tissue can be revealed by injecting fluorochrome-conjugated lectins subcutaneously. Moreover, the in vivo labeling approach provides a way to evaluate lymphatic vessel density and functions. Coupled to blood vessel, adipocyte and immune cell staining, the protocol allows for high-resolution mapping of the subcutaneous adipose tissue by 3D imaging.


Measurement of Interpeptidic CuII Exchange Rate Constants of CuII-Amyloid-β Complexes to Small Peptide Motifs by Tryptophan Fluorescence Quenching.
The interpeptidic CuII exchange rate constants were measured for two Cu amyloid-β complexes, Cu(Aβ1-16) and Cu(Aβ1-28), to fluorescent peptides GHW and DAHW using a quantitative tryptophan fluorescence quenching methodology. The second-order rate constants were determined at three pH values (6.8, 7.4, and 8.7) important to the two Cu(Aβ) coordination complexes, components Cu(Aβ)I and Cu(Aβ)II. The interpeptidic CuII exchange rate constant is approximately 104 M-1 s-1 but varies in magnitude depending on many variables. These include pH, length of the Aβ peptide, location of the anchoring histidine ligand in the fluorescent peptide, number of amide deprotonations required in the tryptophan peptide to coordinate CuII, and interconversion between Cu(Aβ)I and Cu(Aβ)II. We also present EPR data probing the CuII exchange between peptides and the formation of ternary species between Cu(Aβ) and GHW. As the nonfluorescent GHK and DAHK peptides are important motifs found in the blood and serum, their ability to sequester CuII ions from Cu(Aβ) complexes may be relevant for the metal homeostasis and its implication in Alzheimer's disease. Thus, their kinetic CuII interpeptidic exchange rate constants are important chemical rate constants that can help elucidate the complex CuII trafficking puzzle in the synaptic cleft.


Differential regulation of the fiber type-specific gene expression of the sarcoplasmic reticulum calcium-ATPase isoforms induced by exercise training.
The regulatory role of adenosine monophosphate-activated protein kinase (AMPK)-α2 on sarcoplasmic reticulum calcium-ATPase (SERCA) 1a and SERCA2a in different skeletal muscle fiber types has yet to be elucidated. Sedentary (Sed) or exercise-trained (Ex) wild-type (WT) and AMPKα2-kinase dead (KD) transgenic mice, which overexpress a mutated and inactivated AMPKα2 subunit, were utilized to characterize how genotype or exercise training influenced the regulation of SERCA isoforms in gastrocnemius. As expected, both Sed and Ex KD mice had >40% lower AMPK phosphorylation and 30% lower SERCA1a protein than WT mice (P < 0.05). In contrast, SERCA2a protein was not different among KD and WT mice. Exercise increased SERCA1a and SERCA2a protein content among WT and KD mice, compared with their Sed counterparts. Maximal SERCA activity was lower in KD mice, compared with WT. Total phospholamban protein was higher in KD mice than in WT and lower in Ex compared with Sed mice. Exercise training increased phospholamban Ser(16) phosphorylation in WT mice. Laser capture microdissection and quantitative PCR indicated that SERCA1a mRNA expression among type I fibers was not altered by genotype or exercise, but SERCA2a mRNA was increased 30-fold in WT+Ex, compared with WT+Sed. In contrast, the exercise-stimulated increase for SERCA2a mRNA was blunted in KD mice. Exercise upregulated SERCA1a and SERCA2a mRNA among type II fibers, but was not altered by genotype. Collectively, these data suggest that exercise differentially influences SERCA isoform expression in type I and type II fibers. Additionally, AMPKα2 influences the regulation of SERCA2a mRNA in type I skeletal muscle fibers following exercise training.


Sellar inflammatory mass with inflammatory bowel disease.
Inflammatory bowel disease may be associated with different intracranial disorders. An inflammatory sellar mass is very rare but includes a variety of noninfectious causes including lymphocytic hypophysitis, granulomatous inflammation and Wegener's granulomatosis. A 32-year-old man was diagnosed with an inflammatory sellar mass associated with an extensive colonic inflammatory process clinically characteristic of Crohn's disease. The concurrent onset of these inflammatory disorders in distinctly separate sites may reflect their common embryological origin or represent an unusual form of metastatic Crohn's disease. Further studies are needed to determine if less overt or focal sellar inflammatory processes occur in inflammatory bowel disease, particularly in Crohn's disease because their occurrence may be critically relevant for long-term management.


The parabasalid symbiont community of Heterotermes aureus: Molecular and morphological characterization of four new species and reestablishment of the genus Cononympha.
The subterranean termite Heterotermes aureus is endemic to arid regions of southwestern USA and northern Mexico. Like other termites in the family Rhinotermitidae, it harbors a community of protists (Phylum Parabasalia) in its hindgut that aid in cellulose digestion. We investigated the hindgut community of H. aureus using light microscopy, single cell isolation, and high throughput amplicon sequencing. Here we describe four new parabasalid species from the classes Trichonymphea and Spirotrichonymphea. Three of the new species include Pseudotrichonympha aurea (Trichonymphea), Holomastigotoides aureus, and Holomastigotoides oxyrhynchus (Spirotrichonymphea). The fourth new species is a Spirotrichonympha-like protist for which we reinstate the genus Cononympha and describe under the name Cononympha aurea (Spirotrichonymphea). We also used high throughput amplicon sequencing with custom primers on DNA from fresh and ethanol preserved termites collected across the southwest USA and Mexico to investigate population-level differences in hindgut community composition. We report that the community is highly similar across populations: no additional parabasalid species were identified in any of the H. aureus specimens, but several specimens appeared to lack either C. aurea or H. oxyrhynchus.


Wagner apparatus: a portable traction device for the femur.
The Wagner device has proved to be an extremely useful tool for controlling difficult femoral fractures. The fracture and soft tissue injury can be adequately controlled and the patient mobilized by use of the traveling traction principle. This apparatus is easily applied, and the fracture length and alignment can be readily maintained. The device has proven useful in permitting wound care and control. In the complicated polytraumatized patient it facilitates improved pulmonary care and patient hygiene. The advantage the Wagner device affords over conventional traction devices and other external fixation units makes its use a viable orthopaedic alternative.


Liver × receptor ligands disrupt breast cancer cell proliferation through an E2F-mediated mechanism.
Liver × receptors (LXRs) are members of the nuclear receptor family of ligand-dependent transcription factors and have established functions as regulators of cholesterol, glucose, and fatty acid metabolism and inflammatory responses. Published reports of anti-proliferative effects of synthetic LXR ligands on breast, prostate, ovarian, lung, skin, and colorectal cancer cells suggest that LXRs are potential targets in cancer prevention and treatment. To further determine the effects of LXR ligands and identify their potential mechanisms of action in breast cancer cells, we carried out microarray analysis of gene expression in four breast cancer cell lines following treatments with the synthetic LXR ligand GW3965. Differentially expressed genes were further subjected to gene ontology and pathway analyses, and their expression profiles and associations with disease parameters and outcomes were examined in clinical samples. Response of E2F target genes were validated by real-time PCR, and the posited role of E2F2 in breast cancer cell proliferation was tested by RNA interference experiments. We observed cell line-specific transcriptional responses as well as a set of common responsive genes. In the common responsive gene set, upregulated genes tend to function in the known metabolic effects of LXR ligands and LXRs whereas the downregulated genes mostly include those which function in cell cycle regulation, DNA replication, and other cell proliferation-related processes. Transcription factor binding site analysis of the downregulated genes revealed an enrichment of E2F binding site sequence motifs. Correspondingly, E2F2 transcript levels are downregulated following LXR ligand treatment. Knockdown of E2F2 expression, similar to LXR ligand treatment, resulted in a significant disruption of estrogen receptor positive breast cancer cell proliferation. Ligand treatment also decreased E2F2 binding to cis-regulatory regions of target genes. Hierarchical clustering of breast cancer patients based on the expression profiles of the commonly downregulated LXR ligand-responsive genes showed a strong association of these genes with patient survival. Taken together, these results indicate that LXR ligands target gene networks, including those regulated by E2F family members, are critical for tumor biology and disease progression and merit further consideration as potential agents in the prevention and treatment of breast cancers.


Orthostatic hypotension, orthostatic hypertension, and ambulatory blood pressure in patients with chronic kidney disease in CRIC.
Orthostatic changes in blood pressure (BP), either orthostatic hypotension or orthostatic hypertension (OHTN), are common among patients with chronic kidney disease. Whether they are associated with unique out-of-office BP phenotypes is unknown. CRIC is a prospective, multicenter, observational cohort study of participants with CKD. BP measured at 2 min after standing and ambulatory BP monitoring (ABPM) were obtained on 1386 participants. Orthostatic hypotension was defined as a 20 mmHg drop in SBP or 10 mmHg drop in DBP when changing from seated to standing positions. Systolic and diastolic night-to-day ratio was also calculated. OHTN was defined as a 20 or 10 mmHg rise in SBP or DBP when changing from a seated to a standing position. White-coat effect (WCE) was defined as seated minus daytime ambulatory BP. Of the 1386 participants (age: 58 ± 10 years, 44% female, 39% black), 68 had orthostatic hypotension and 153 had OHTN. Postural reduction in SBP or DBP was positively associated with greater systolic and diastolic WCE and systolic and diastolic night-to-day ratio. Orthostatic hypotension was positively associated with diastolic WCE (β = 3 [0.2, 5.9]). Diastolic OHTN was negatively associated with systolic WCE (β = -4 [-7.2, -0.5]) and diastolic WCE (β = -6 [-8.1, -4.2]). Postural change in BP was associated with WCE and night-to-day-ratio. Orthostatic hypotension was positively associated with WCE and OHTN was negatively associated with WCE. These findings strengthen observations that postural changes in BP may associate with distinct BP patterns throughout the day. These observations are informative for subsequent research tailoring orthostatic hypotension and OHTN treatment to specific BP phenotypes.


Heterogeneity of mucus glycoproteins from cystic fibrotic sputum. Are there different families of mucins?
High-Mr mucin glycopeptides prepared from sputum of an individual with cystic fibrosis (CF) were studied by ion-exchange h.p.l.c. The glycopeptides were heterogeneous and a number of partially resolved populations were identified. Whole mucins from the gel phase were separated into four fractions by isopycnic density-gradient centrifugation in CsCl, and high-Mr glycopeptides from these fractions were examined by ion-exchange h.p.l.c. The acidic nature of the high-Mr glycopeptides increased with increasing buoyant density of the intact mucins, and a periodate-Schiff (PAS)-rich and an extremely high-iron diamine (HID)-reactive component were present in the lowest and highest density fractions respectively. The various glycopeptide populations were identified in different proportions in mucins from four other individuals with CF. CF sputum thus seems to contain distinct mucin populations containing different oligosaccharide clusters corresponding to these high-Mr glycopeptides.


A national outbreak of Salmonella enteritidis infections from ice cream. The Investigation Team.
In September 1994, the Minnesota Department of Health detected an increase in the number of reports of Salmonella enteritidis infections. After a case-control study implicated a nationally distributed brand of ice cream (Schwan's) in the outbreak, the product was recalled and further epidemiologic and microbiologic investigations were conducted. We defined an outbreak-associated case of S. enteritidis infection as one in which S. enteritidis was cultured from a person who became ill in September or October 1994. We established national surveillance and surveyed customers of the implicated manufacturer. The steps involved in the manufacture of ice cream associated with cases of S. enteritidis infection were compared with those of products not known to be associated with infection matched for the date of manufacture. Cultures for bacteria were obtained from ice cream samples, the ice cream plant, and tanker trailers that had transported the ice cream base (premix) to the plant. We estimate that S. enteritidis gastroenteritis developed in 224,000 persons in the United States after they ate Schwan's ice cream. The attack rate for consumers was 6.6 percent. Ice cream associated with infection contained a higher percentage of premix that had been transported by tanker that had carried nonpasteurized eggs immediately before (P = 0.02). S. enteritidis was isolated from 8 of 226 ice cream products (3 percent), but not from environmental samples obtained from the ice cream plant (n = 157) or tanker trailers (n = 204). This nationwide outbreak of salmonellosis was most likely the result of contamination of pasteurized ice cream premix during transport in tanker trailers that had previously carried nonpasteurized liquid eggs containing S. enteritidis. To prevent further outbreaks, food products not destined for repasteurization should be transported in dedicated containers.


Environment: a perspective of the self-care deficit nursing theory.
In this column the author presents an examination of Orem's conceptualization of environment as a metaparadigm concept, the role of environment in the theoretical structure of the self-care deficit nursing theory, and the creation of a developmental environment as one of the ways of helping another. Suggestions for further work related to the conceptualization of environment and the organization of existent knowledge are provided. Application of the concept of nursing agency and the environmental factor of healthcare system illustrate the utility of self-care deficit nursing theory in relation to the recommendations for changes in nursing education and for redesign of the healthcare system.


Towards modular bone tissue engineering using Ti-Co-doped phosphate glass microspheres: cytocompatibility and dynamic culture studies.
The production of large quantities of functional vascularized bone tissue ex vivo still represent an unmet clinical challenge. Microcarriers offer a potential solution to scalable manufacture of bone tissue due to their high surface area-to-volume ratio and the capacity to be assembled using a modular approach. Microcarriers made of phosphate bioactive glass doped with titanium dioxide have been previously shown to enhance proliferation of osteoblast progenitors and maturation towards functional osteoblasts. Furthemore, doping with cobalt appears to mimic hypoxic conditions that have a key role in promoting angiogenesis. This characteristic could be exploited to meet the clinical requirement of producing vascularized units of bone tissue. In the current study, the human osteosarcoma cell line MG-63 was cultured on phosphate glass microspheres doped with 5% mol titanium dioxide and different concentrations of cobalt oxide (0%, 2% and 5% mol), under static and dynamic conditions (150 and 300 rpm on an orbital shaker). Cell proliferation and the formation of aggregates of cells and microspheres were observed over a period of two weeks in all glass compositions, thus confirming the biocompatibility of the substrate and the suitability of this system for the formation of compact micro-units of tissue. At the concentrations tested, cobalt was not found to be cytotoxic and did not alter cell metabolism. On the other hand, the dynamic environment played a key role, with moderate agitation having a positive effect on cell proliferation while higher agitation resulting in impaired cell growth. Finally, in static culture assays, the capacity of cobalt doping to induce vascular endothelial growth factor (VEGF) upregulation by osteoblastic cells was observed, but was not found to increase linearly with cobalt oxide content. In conclusion, Ti-Co phosphate glasses were found to support osteoblastic cell growth and aggregate formation that is a necessary precursor to tissue formation and the upregaulation of VEGF production can potentially support vascularization.


Methyl green-pyronin; basis of selective staining of nucleic acids.
1. Methyl green stains selectively highly polymerized desoxyribonucleic acid, and fails to stain, to any significant extent, depolymerized desoxyribonucleic acid and ribonucleic acid. 2. Pyronin stains preferentially low polymers of nucleic acid. 3. Triphenylmethane dyes with two amino groups appear to share the selectivity of methyl green. Those with three amino groups are not selective. 4. A stereochemical hypothesis is offered to account for these observations.


The Interplay Between Cholesterol and Amyloid-β on HT22 Cell Viability, Morphology, and Receptor Tyrosine Kinase Signaling.
There is a lack of understanding in the molecular and cellular mechanisms of Alzheimer's disease that has hindered progress on therapeutic development. The focus has been on targeting toxic amyloid-β (Aβ) pathology, but these therapeutics have generally failed in clinical trials. Aβ is an aggregation-prone protein that has been shown to disrupt cell membrane structure in molecular biophysics studies and interfere with membrane receptor signaling in cell and animal studies. Whether the lipid membrane or specific receptors are the primary target of attack has not been determined. This work elucidates some of the interplay between membrane cholesterol and Aβ42 on HT22 neuronal cell viability, morphology, and platelet-derived growth factor (PDGF) signaling pathways. The effects of cholesterol depletion by methyl-β-cyclodextrin followed by treatment with Aβ and/or PDGF-AA were assessed by MTT cell viability assays, western blot, optical and AFM microscopy. Cell viability studies show that cholesterol depletion was mildly protective against Aβ toxicity. Together cholesterol reduction and Aβ42 treatment compounded the disruption of the PDGFα receptor activation. Phase contrast optical microscopy and live cell atomic force microscopy imaging revealed that cytotoxic levels of Aβ42 caused morphological changes including cell membrane damage, cytoskeletal disruption, and impaired cell adhesion; cell damage was ameliorated by cellular cholesterol depletion. Cholesterol depletion impacted the effects of Aβ42 on HT22 cell viability, morphology, and receptor tyrosine kinase signaling.


Present and future are getting confused: are we equipped to face the technological revolution?
This is a commentary reporting the outcome of a workshop promoted by the Department of Woman's and Child's Health of the University of Padua (Italy) focused on the emerging issue of what seems to be the increasing agemone role of technology. Over the centuries, technology has always been at the service of science, with theoretical insights anticipating experimental proofs. Over the last decades, however, the situation has radically changed, due to several factors. Technology seems to be playing an agemone role. The present and notably the future generation of scientists have major challenges to face. They have to deal with the forces generated by the infosphera; to dominate the technology and to maintain the capacity of generating inquisitive, creative, ethical and spiritual thoughts capable of addressing new scientific hypotheses and projects directed to the individual and collective good. However, in this scenario, what seems more relevant is to focus all our efforts in preparing ourselves, first, and then the new generations to face these challenges. From this point of view, the academic institutions and the scientific societies, have a major responsibility to deal with. The academic ecosystem traditionally used to educate the new generation of professionals as well as, and most importantly, the cultural, the professional pathways presently used to form them need to be extensively revised. The time is running short and the stakes are high. The debate is open.


Endochondral bone formation from hydrogel carriers loaded with BMP2-transduced cells.
The success of ex vivo viral gene therapy systems for promoting bone formation could be improved through the development of systems to spatially localize gene expression. Towards this goal, we have encapsulated adenovirus-transduced human diploid fetal lung fibroblasts (MRC-5) expressing bone morphogenetic protein-type 2 (BMP-2) within non-degradable poly(ethylene glycol)-diacrylate (PEG-DA) hydrogels and implanted these intramuscularly to promote endochondral bone formation. To optimize BMP-2 secretion, the molecular weight of the polymers and cell densities were varied. Polymers with molecular weights of 6, 10, and 20 kDa were used to prepare hydrogels containing 1, 5, or 10 million transduced cells. The results showed that 10 million transduced fibroblasts that was the maximum number of cells feasible for encapsulation within PEG-DA 10 and 20 kDa hydrogels produced the highest amount of secreted BMP-2 protein. Encapsulation of MRC-5 and transduced fibroblasts resulted in 71 and 58% cell viability, respectively. The bioactivity of secreted BMP-2 protein from the hydrogels was confirmed with an alkaline phosphatase assay. Micro-CT of the lower limb muscles of NOD/SCID mice following implantation with hydrogels showed 39.5 +/- 25.0 mm3 mineralized tissue and 31.8 +/- 7.8 mm3 for the cell-injected mice, and the bone was localized to the hydrogel surfaces. Histology revealed bone as well as cartilage for both hydrogel implanted and cell-injected animals.


Beta-cell response during a meal test: a comparative study of incremental doses of repaglinide in type 2 diabetic patients.
To assess the effects of incremental doses of repaglinide on postprandial insulin and glucose profiles after a standard 500-kcal test meal. Sixteen diet-treated Caucasians with type 2 diabetes (mean HbA(1c) 8.4%) were enrolled in this randomized, open-label, crossover trial. Subjects received 0.5, 1, 2, and 4 mg repaglinide or placebo in a random fashion, followed by a standard 500-kcal test meal on 5 separate study days, 1 week apart. The insulinogenic index (DeltaI30/DeltaG30) and insulin area under the curve (AUC) from 0 to 30 min (AUC(0-30)) were higher with the 4-mg drug dose compared with the two lower doses and with 2 mg compared with 0.5 mg. On subgroup analysis, the incremental insulin responses were apparent only in the fasting plasma glucose (FPG) < 9-mmol/l subgroup of subjects and not in the FPG >9-mmol/l subgroup. There was a significant dose-related increase in the late postprandial insulin secretion (insulin AUC(120-240)), which resulted in hypoglycemia in four subjects. Proinsulin-to-insulin ratios at 30 and 60 min improved with increasing doses of repaglinide; higher drug doses (2 and 4 mg) were more effective than the 0.5- and 1-mg doses. Significant dose-related increases in early insulin secretion were found only in less advanced diabetic subjects. In advanced diabetic patients, only the maximum dose (4 mg) was significant compared with placebo. Better proinsulin-to-insulin processing was noted with increasing drug doses.


Multicenter observational prehospital resuscitation on helicopter study.
Earlier use of in-hospital plasma, platelets, and red blood cells (RBCs) has improved survival in trauma patients with severe hemorrhage. Retrospective studies have associated improved early survival with prehospital blood product transfusion (PHT). We hypothesized that PHT of plasma and/or RBCs would result in improved survival after injury in patients transported by helicopter. Adult trauma patients transported by helicopter from the scene to nine Level 1 trauma centers were prospectively observed from January to November 2015. Five helicopter systems had plasma and/or RBCs, whereas the other four helicopter systems used only crystalloid resuscitation. All patients meeting predetermined high-risk criteria were analyzed. Patients receiving PHT were compared with patients not receiving PHT. Our primary analysis compared mortality at 3 hours, 24 hours, and 30 days, using logistic regression to adjust for confounders and site heterogeneity to model patients who were matched on propensity scores. Twenty-five thousand one hundred eighteen trauma patients were admitted, 2,341 (9%) were transported by helicopter, of which 1,058 (45%) met the highest-risk criteria. Five hundred eighty-five of 1,058 patients were flown on helicopters carrying blood products. In the systems with blood available, prehospital median systolic blood pressure (125 vs 128) and Glasgow Coma Scale (7 vs 14) was significantly lower, whereas median Injury Severity Score was significantly higher (21 vs 14). Unadjusted mortality was significantly higher in the systems with blood products available, at 3 hours (8.4% vs 3.6%), 24 hours (12.6% vs 8.9%), and 30 days (19.3% vs 13.3%). Twenty-four percent of eligible patients received a PHT. A median of 1 unit of RBCs and plasma were transfused prehospital. Of patients receiving PHT, 24% received only plasma, 7% received only RBCs, and 69% received both. In the propensity score matching analysis (n = 109), PHT was not significantly associated with mortality at any time point, although only 10% of the high-risk sample were able to be matched. Because of the unexpected imbalance in systolic blood pressure, Glasgow Coma Scale, and Injury Severity Score between systems with and without blood products on helicopters, matching was limited, and the results of this study are inconclusive. With few units transfused to each patient and small outcome differences between groups, it is likely large, multicenter, randomized studies will be required to detect survival differences in this important population. Level II.


Antitumor effect of lung cancer vaccine with umbilical blood dendritic cells in reconstituted SCID mice.
Dendritic cells (DCs) are important cells in initiating an immune response. A generation of functional DCs has potential clinical use in treating cancer. However, the source of DCs and patient immunodeficiency with cancer have been hindrances in clinical therapy. We generated DCs from human umbilical cord blood mononuclear cells (UBMCs) with recombinant human granulocyte-macrophage colony stimulating factor, recombinant human interleukin-4, and recombinant human tumor necrosis factor-alpha. The mature DC-A549 lung cancer vaccine (AgL-DC) was prepared through loading A549 lysate, treating with lipopolysaccharide (LPS) and positive selecting with CD83 magnetic beads. AgL-DC can secrete interleukin (IL)-12 and IL-1. Further in vitro analysis showed that AgL-DC notably induced human UBMC lymphocyte proliferation (p < 0.01) by 3-(4,5-dimethylthiazol-z-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, increased the cytotoxic T-lymphocyte (CTL) activity of UBMC lymphocytes against A549 cells (p < 0.05, at effector cells:target cells ratios of 50:1 and 100:1) by lactate dehydrogenase (LDH) cytotoxic assay, and improved production of IL-6 and tumor necrosis factor-beta (p < 0.01, p < 0.05) by enzyme-linked immunosorbent assay. Subsequently, the reconstitute immunity model in severe combined immunodeficiencies (SCID) mice has been established using human UBMC transplantation, and similar trends to results of UBMC in vitro experiments have been shown in lymphocyte proliferation, CTL activity, and IL-6 and tumor necrosis factor-beta secretion levels in these models. AgL-DC also significantly (p < 0.01) increased the antitumor effect in vivo. The tumor infiltrating immunocytes were positively expressed human CD83 and CD3 molecules, and they were negatively expressed in tumor tissue treated with control. These results have demonstrated that umbilical cord DCs are a useful source of vaccine cells for augmenting CTL-mediated cytotoxicity and have potential usefulness in cellular therapy for human cancer in a new vaccination strategy.


Synthetic 3D PEG-Anisogel Tailored with Fibronectin Fragments Induce Aligned Nerve Extension.
An enzymatically cross-linked polyethylene glycol (PEG)-based hydrogel was engineered to promote and align nerve cells in a three-dimensional manner. To render the injectable, otherwise bioinert, PEG-based material supportive for cell growth, its mechanical and biochemical properties were optimized. A recombinant fibronectin fragment (FNIII9*-10/12-14) was coupled to the PEG backbone during gelation to provide cell adhesive and growth factor binding domains in close vicinity. Compared to full-length fibronectin, FNIII9*-10/12-14 supports nerve growth at similar concentrations. In a 3D environment, only the ultrasoft 1 w/v% PEG hydrogels with a storage modulus of ∼10 Pa promoted neuronal growth. This gel was used to establish the first fully synthetic, injectable Anisogel by the addition of magnetically aligned microelements, such as rod-shaped microgels or short fibers. The Anisogel led to linear neurite extension and represents a large step in the direction of clinical translation with the opportunity to treat acute spinal cord injuries.


Development of non-transgenic glyphosate tolerant wheat by TILLING.
Glyphosate (N-phosphonomethyl-glycine) is the world's most widely used broad spectrum, post-emergence herbicide. It inhibits the chloroplast-targeted enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS; EC 18.104.22.168), a component of the plant and microorganism-specific shikimate pathway and a key catalyst in the production of aromatic amino acids. Variants of EPSPS that are not inhibited by glyphosate due to particular amino acid alterations in the active site of the enzyme are known. Some of these variants have been identified in weed species that have developed resistance to glyphosate because of the strong selective pressure of continuous, heavy glyphosate use. We have used TILLING (Targeting Induced Local Lesions in Genomes), a non-transgenic, target-selected, reverse genetics, mutation breeding technique, and conventional genetic crosses, to identify and combine, through two rounds of mutagenesis, wheat lines having both T102I and P106S (so-called TIPS enzyme) mutations in both the A and the D sub-genome homoeologous copies of the wheat EPSPS gene. The combined effects of the T102I and P106S mutations are known from previous work in multiple species to minimize the binding of the herbicide while maintaining the affinity of the catalytic site for its native substrates. These novel wheat lines exhibit substantial tolerance to commercially relevant levels of glyphosate.


Health Benefits of Fermented Bamboo Shoots: The Twenty-First Century Green Gold of Northeast India.
The word "bamboo" reminds us of "a hollow stick," but it is filled with a plethora of health benefits. The tribals of northeastern India ferment these beneficial bamboo shoots for the goodness of mankind. Fermentation is an important age-old biotechnological procedure used for the preservation of food products. Fermented bamboo shoots form the niche for many microorganisms, and this confers positive effects and advantages in many ways. These magical shoots have tremendous health benefits like anti-cancer, anti-oxidant, anti-aging, cardioprotective, weight loss, probiotics, to name a few. Apart from health benefits, fermented bamboo shoots form important functional foods and have industrial and economical values. Though these are commonly found and started in the tribal area, and local markets, today, they are valuable all around the world, as popular as gold. Hence, fermented bamboo shoots are referred as "green gold" of India. This review briefs about various health benefits, advantages, disadvantages, future scope, and finally the economic values of fermented bamboo shoots, the "green gold" of the twenty-first century.


Effect of transported segment size on the new bone formation of the rabbit femur in the Ilizarov bone transport method.
This study aimed to compare histological and radiological union in the bone transport of 3 segments of different sizes to reconstruct the rabbit femur's bone defects. Thirty rabbits were divided into 3 groups; a 1-cm defect was created in the femur in all rabbits. The length of the segment to be transferred was 10% of the femur length in group 1, 15% in group 2, and 20% in group 3. All defects were reconstructed by applying bone transport. At the end of the consolidation period, the distraction zone was compared radiologically and histologically. While there was no radiological difference between the groups, the highest histological scores were obtained from group 3. Osteocalcin staining revealed similar involvement in groups 2 and 3, butless involvement in group 1. Evidence from this study has shown that as the size of the segment used for bone transport increases, more stable fixation and better histological union tissue can be obtained in the rabbit femoral defect model.


The relationship between attachment style and placement of parents in adults' attachment networks over time.
Using a bull's-eye hierarchical mapping technique (HMT), the present study examined placement of parents in adults' attachment networks over time. We hypothesized that attachment style would predict distance at which network members (mother, father, and romantic partner) would be placed from the core-self over time. Participants completed the HMT on two occasions, 12 months apart. Concurrently and over time, fathers were placed further from the core-self than mothers. Attachment style explained unique variance, beyond that accounted for by individual and relationship characteristics. Specifically, network members with whom participants reported greater attachment insecurity were placed further from the core-self concurrently. Mothers with whom participants reported greater attachment insecurity were placed further from the core-self over time. Unsatisfactory attachment relationships with father and partner and those marked by higher attachment insecurity were more likely to be excluded from attachment networks over time. Findings suggest that attachment style, relationship quality, romantic relationship status, and parents' marital status determine the placement of parents in adults' attachment networks.


Age-related variation in health status after age 60.
Disability, functionality, and morbidity are often used to describe the health of the elderly. Although particularly important when planning health and social services, knowledge about their distribution and aggregation at different ages is limited. We aim to characterize the variation of health status in a 60+ old population using five indicators of health separately and in combination. 3080 adults 60+ living in Sweden between 2001 and 2004 and participating at the SNAC-K population-based cohort study. Health indicators: number of chronic diseases, gait speed, Mini Mental State Examination (MMSE), disability in instrumental-activities of daily living (I-ADL), and in personal-ADL (P-ADL). Probability of multimorbidity and probability of slow gait speed were already above 60% and 20% among sexagenarians. Median MMSE and median I-ADL showed good performance range until age 84; median P-ADL was close to zero up to age 90. Thirty% of sexagenarians and 11% of septuagenarians had no morbidity and no impairment, 92% and 80% of them had no disability. Twenty-eight% of octogenarians had multimorbidity but only 27% had some I-ADL disability. Among nonagenarians, 13% had severe disability and impaired functioning while 12% had multimorbidity and slow gait speed. Age 80-85 is a transitional period when major health changes take place. Until age 80, most people do not have functional impairment or disability, despite the presence of chronic disorders. Disability becomes common only after age 90. This implies an increasing need of medical care after age 70, whereas social care, including institutionalization, becomes a necessity only in nonagenarians.


Phenotypic checklist to screen for fragile X syndrome in people with mental retardation.
The development of a phenotypic checklist for identifying people with fragile X syndrome is described. The checklist was designed to identify people with developmental disabilities of unknown causes for molecular genetic testing for fragile X syndrome. The list consists of 28 items (7 on physical characteristics and 21 on behavioral features). Validation data were collected for 110 boys and men with fragile X syndrome and for 79 members of a control group, matched for CA, level of cognitive development, and social (mal)adaptation. On the basis of checklist results, those boys who are likely to be diagnosed as having fragile X syndrome can be identified. The screening list can be considered to be a consistent, reliable, and valid instrument.


Hospitalizations for asthma in children are linked to undertreatment and insufficient asthma education.
Most hospital admissions for asthma exacerbation are avoidable with adequate disease management. The objective of this study was to describe the characteristics of children hospitalized with an asthma exacerbation to identify modifiable factors leading to hospitalization. The study was conducted in 14 pediatric units and included children 3-17 years of age who were hospitalized for an asthma exacerbation. The present analysis covers 498 children with known asthma. Staff physicians used a standardized questionnaire to collect data. Asthma history came from a parental interview and included usual asthma care, frequency of symptoms and quick-relief medication use in the previous month, frequency of exacerbations and number of unscheduled healthcare visits during the past year, and prior asthma-related hospitalizations. More than half the children had previously been hospitalized for an exacerbation, 42% used continuous inhaled corticosteroids, and 57% had a regular follow-up for asthma. Asthma had been well controlled over the past year for 11%, 12% had experienced exacerbations during the past year but that had been optimally controlled during the previous month, and 11% had recently become poorly controlled (infrequent exacerbations in the previous year and non-optimal control in the previous month). The remaining 327 children (66%) were consistently poorly controlled (non-optimal asthma control in the previous month and frequent exacerbations over the previous year). Among this group, 69% had at least one of the following preventable risk factors for hospitalization: no regular controller therapy (49%), no asthma action plan (40%), or no follow-up for asthma (35%). Two-thirds of the children with asthma hospitalized for an exacerbation had been consistently poorly controlled during the previous year. They were frequently undertreated and insufficiently educated about asthma. Further efforts are needed to improve asthma treatment and education in France.


Posttraumatic Parotid Sialocele Complicating a Mandibular Fracture: A Case Report.
Blunt trauma to the parotid resulting in the formation of a sialocele is rare, with only three cases identified in the literature. We present a unique case involving a 32-year-old man with blunt trauma resulting in a left mandibular angle fracture. The patient underwent open reduction and internal fixation of the left mandibular angle fracture via transoral approach. At follow-up, after resolution of the edema from the injury, a sialocele was noted in the region of the left anterior parotid gland. The patient was treated conservatively with antisialagogues, pressure dressings, and multiple percutaneous aspirations that ultimately resulted in resolution of the sialocele.


Redox State in Atrial Fibrillation Pathogenesis and Relevant Therapeutic Approaches.
Myocardial redox state is a critical determinant of atrial biology, regulating cardiomyocyte apoptosis, ion channel function, and cardiac hypertrophy/fibrosis and function. Nevertheless, it remains unclear whether the targeting of atrial redox state is a rational therapeutic strategy for atrial fibrillation prevention. To review the role of atrial redox state and anti-oxidant therapies in atrial fibrillation. Published literature in Medline was searched for experimental and clinical evidence linking myocardial redox state with atrial fibrillation pathogenesis as well as studies looking into the role of redoxtargeting therapies in the prevention of atrial fibrillation. Data from animal models have shown that altered myocardial nitroso-redox balance and NADPH oxidases activity are causally involved in the pathogenesis of atrial fibrillation. Similarly experimental animal data supports that increased reactive oxygen / nitrogen species formation in the atrial tissue is associated with altered electrophysiological properties of atrial myocytes and electrical remodeling, favoring atrial fibrillation development. In humans, randomized clinical studies using redox-related therapeutic approaches (e.g. statins or antioxidant agents) have not documented any benefits in the prevention of atrial fibrillation development (mainly post-operative atrial fibrillation risk). Despite strong experimental and translational data supporting the role of atrial redox state in atrial fibrillation pathogenesis, such mechanistic evidence has not been translated to clinical benefits in atrial fibrillation risk in randomized clinical studies using redox-related therapies.


Sleeping Beauty transposon mutagenesis identifies genes that cooperate with mutant Smad4 in gastric cancer development.
Mutations in SMAD4 predispose to the development of gastrointestinal cancer, which is the third leading cause of cancer-related deaths. To identify genes driving gastric cancer (GC) development, we performed a Sleeping Beauty (SB) transposon mutagenesis screen in the stomach of Smad4(+/-) mutant mice. This screen identified 59 candidate GC trunk drivers and a much larger number of candidate GC progression genes. Strikingly, 22 SB-identified trunk drivers are known or candidate cancer genes, whereas four SB-identified trunk drivers, including PTEN, SMAD4, RNF43, and NF1, are known human GC trunk drivers. Similar to human GC, pathway analyses identified WNT, TGF-β, and PI3K-PTEN signaling, ubiquitin-mediated proteolysis, adherens junctions, and RNA degradation in addition to genes involved in chromatin modification and organization as highly deregulated pathways in GC. Comparative oncogenomic filtering of the complete list of SB-identified genes showed that they are highly enriched for genes mutated in human GC and identified many candidate human GC genes. Finally, by comparing our complete list of SB-identified genes against the list of mutated genes identified in five large-scale human GC sequencing studies, we identified LDL receptor-related protein 1B (LRP1B) as a previously unidentified human candidate GC tumor suppressor gene. In LRP1B, 129 mutations were found in 462 human GC samples sequenced, and LRP1B is one of the top 10 most deleted genes identified in a panel of 3,312 human cancers. SB mutagenesis has, thus, helped to catalog the cooperative molecular mechanisms driving SMAD4-induced GC growth and discover genes with potential clinical importance in human GC.


How soft is that pillow? The perceptual localization of the hand and the haptic assessment of contact rigidity.
A new haptic illusion is described, in which the location of the mobile object affects the perception of its rigidity. There is theoretical and experimental support for the notion that limb position sense results from the brain combining ongoing sensory information with expectations arising from prior experience. How does this probabilistic state information affect one's tactile perception of the environment mechanics? In a simple estimation process, human subjects were asked to report the relative rigidity of two simulated virtual objects. One of the objects remained fixed in space and had various coefficients of stiffness. The other virtual object had constant stiffness but moved with respect to the subjects. Earlier work suggested that the perception of an object's rigidity is consistent with a process of regression between the contact force and the perceived amount of penetration inside the object's boundary. The amount of penetration perceived by the subject was affected by varying the position of the object. This, in turn, had a predictable effect on the perceived rigidity of the contact. Subjects' reports on the relative rigidity of the object are best accounted for by a probabilistic model in which the perceived boundary of the object is estimated based on its current location and on past observations. Therefore, the perception of contact rigidity is accounted for by a stochastic process of state estimation underlying proprioceptive localization of the hand.


Molecular mechanisms regulating immune responses in thromboangiitis obliterans: A comprehensive review.
Thromboangiitis obliterans (TAO) is a thrombotic-occlusive as well as an inflammatory peripheral vascular disease with unknown etiology. Recent evidence has supported the immunopathogenesis of the disease, however, the factors contributing to the altered immune function and vascular tissue inflammation are still unclear. This review was intended to collate the more current knowledge on the regulatory molecules involved in TAO from an immunoreactive perspective. The homeostasis of the immune system as well as a variety of progenitor cell populations appear to be affected during TAO and these alterations are associated with intrinsic signaling defects that are directing to an improved understanding of the crosstalk between angiogenesis and the immune system, as well as the potential of new co-targeting strategies applying both immunotherapy and angiogenic therapy.


Development of a versatile cell force transducer using moiré mechanism.
Cells alter their shape and morphology and interact with their surrounding environment. Mechanical forces developed by cells to their surrounding environments are fundamental to many physiological processes, such as cell growth, division, migration, and apoptosis. In this paper, a novel optical moiré based biomechanol force sensor was developed for cell traction force mapping. We utilized coherent laser beams to illuminate periodic polymeric substrates where isolated cells were cultured. We demonstrated one-dimensional and two-dimensional traction force mapping via optical moiré for both cardiac myocytes and vascular smooth muscle cells. The magnification effect of the moiré fringe pattern permits a real time monitoring of the mechanical interaction between isolated cells and their underlying periodic polymeric structures.


Aortic Valve Replacement in the Era of Transcatheter Aortic Valve Implantation: Current Status in Korea.
In the era of transcatheter aortic valve implantation, this study was conducted to investigate the current trend of aortic valve procedures in Korea and to evaluate the early and mid-term outcomes of isolated surgical aortic valve replacement (SAVR) using bioprosthetic valves in contemporary Korea. Contemporary outcomes of isolated bioprosthetic SAVR in Korea were analyzed using the datasets on a multicenter basis. Patients who underwent isolated SAVR using bioprostheses from June 2015 to May 2019 were included, and those with concomitant cardiac procedures, SAVR with mechanical valve, or SAVR for infective endocarditis were excluded. A total of 456 patients from 4 large-volume centers were enrolled in this study. Median follow-up duration was 43.4 months. Early postoperative outcomes, mid-term clinical outcomes, and echocardiographic outcomes were evaluated. Mean age of the patients was 73.1 ± 7.3 years, and EuroSCORE II was 2.23 ± 2.09. The cardiopulmonary bypass time and aortic cross-clamp times were median 106 and 76 minutes, respectively. SAVR was performed with full median sternotomy (81.8%), right thoracotomy (14.7%), or partial sternotomy (3.5%). Operative mortality was 1.8%. The incidences of stroke and permanent pacemaker implantation were 1.1% and 1.1%, respectively. Paravalvular regurgitation ≥ mild was detected in 2.6% of the patients. Cumulative incidence of all-cause mortality at 5 years was 13.0%. Cumulative incidences of cardiovascular mortality and bioprosthetic valve dysfunction at 5 years were 7.6% and 6.8%, respectively. The most recent data for isolated SAVR using bioprostheses in Korea resulted in excellent early and mid-term outcomes in a multicenter study.


Factors Influencing Attitude to Menstruation in Elementary School Girls according to the Presence of Menarche.
The purpose of this study was to examine factors related to attitudes to menstruation in elementary school girls according to menarche experience. Data were collected using a structured questionnaire from 208 girls in 5th or 6th grade (90 who were menarche and 118 who were not). The study was a cross-sectional study. Data were analyzed using SPSS/WIN 18.0 program for descriptive statistics, t-test, ANOVA, Pearson correlation, and multiple regression. There were significant differences in body image, menstrual knowledge and attitude between the two groups of students. Results from the multiple regression analysis revealed difference in related factors and in the magnitude of the relationship evaluated by standardized beta coefficients. Significant factors, listed by the magnitude of beta coefficients, among students in menarche were positive affect of menarche, self-oriented perfectionism-critical, negative affect of menarche. Significant factors among non-menarche students were menstrual knowledge and self-oriented perfectionism-critical. There are differences between menarche and non-menarche girls in attitude to menstruation and related factors. Based on the results of the present study, systemic health education according to differences between the two groups should be developed to foster positive attitudes to menstruation.


Insulin action in the double incretin receptor knockout mouse.
The incretins glucagon-like peptide 1 and glucose-dependent insulinotropic polypeptide have been postulated to play a role in regulating insulin action, although the mechanisms behind this relationship remain obscure. We used the hyperinsulinemic-euglycemic clamp to determine sites where insulin action may be modulated in double incretin receptor knockout (DIRKO) mice, which lack endogenous incretin action. DIRKO and wild-type mice were fed regular chow or high-fat diet for 4 months. Clamps were performed on 5-h-fasted, conscious, unrestrained mice using an arterial catheter for sampling. Compared with wild-type mice, chow and high fat-fed DIRKO mice exhibited decreased fat and muscle mass associated with increased energy expenditure and ambulatory activity. Clamp rates of glucose infusion (GIR), endogenous glucose production (endoR(a)), and disappearance (R(d)) were not different in chow-fed wild-type and DIRKO mice, although insulin levels were lower in DIRKO mice. Liver Akt expression was decreased but Akt activation was increased in chow-fed DIRKO compared with wild-type mice. High-fat feeding resulted in fasting hyperinsulinemia and hyperglycemia in wild-type but not in DIRKO mice. GIR, suppression of endoR(a), and stimulation of R(d) were inhibited in high fat-fed wild-type mice but not in DIRKO mice. High-fat feeding resulted in impaired tissue glucose uptake (R(g)) in skeletal muscle of wild-type mice but not of DIRKO mice. Liver and muscle Akt activation was enhanced in high fat-fed DIRKO compared with wild-type mice. In summary, DIRKO mice exhibit enhanced insulin action compared with wild-type mice when fed a regular chow diet and are protected from high-fat diet-induced obesity and insulin resistance.


Serology of pneumocystosis.
Included in this evaluation of serologic tests for American pneumocystis pneumonia is a summary of results reported from an indirect immunofluorescence (IF) test and a comparison of results of the immunodiffusion, complement-fixation, and double diffusion tests. Athough IF was the most sensitive, it dectected only a third of the infections. Specificity of the test was good when serum titers greater than or equal to 1:16 were considered postive. Results of test sera in both European and American laboratories suggested that the low level of measurable antibody in American sera real and not due to technical error. These findings are compatible with the concept that two types of population may be infected with Pneumocystis organisms, one type associated with plasma cell pneumonia and the second with hypoergic pneumocystosis in immunosuppressed patients. Analysis by double diffusion to test antigens revealed their antigenic complexity. Most components, were indentified or partially identified with host elements, and no purely parastic antigens were demonstrated. When cysts isolated from animals and humans were tested, results suggested that they shared at least some common antigens.


BH3 profiling discriminates response to cytarabine-based treatment of acute myelogenous leukemia.
As acute myelogenous leukemia (AML) patient response to cytarabine-based standard-of-care treatment is variable, stratification into subgroups by biomarker-predicted response may lead to improved clinical outcomes. Here, we assess cell mitochondrial depolarization to proapoptotic signaling BH3-only peptides as a surrogate for the function of Bcl-2 family proteins to address clinical response to cytarabine-based therapy in patients with AML (N = 62). Peripheral blood mononuclear cell (PBMC) or bone marrow aspirate specimens were obtained from newly diagnosed patients with AML, viably preserved, and assayed by flow cytometry following BH3 profile assay with individual BH3 peptides. Mann-Whitney analysis indicates biomarker correlation with response to induction therapy: Notably, BIM priming was highly significant (P = 2 × 10(-6)) with a compelling sensitivity/specificity profile [area under curve (AUC) = 0.83; 95% confidence interval (CI), 0.73-0.94; P = 2 × 10(-10)]. Multivariate analysis indicates improved profiles for BIM readout + patient age (AUC = 0.89; 95% CI, 0.81-0.97) and BIM + patient age + cytogenetic status (AUC = 0.91; 95% CI, 0.83-0.98). When patients were stratified by cytogenetic status, BIM readout was significant for both intermediate (P = 0.0017; AUC = 0.88; 95% CI, 0.71-1.04) and unfavorable (P = 0.023; AUC = 0.79; 95% CI, 0.58-1.00) risk groups, demonstrating predictive power independent of cytogenetics. Additional analyses of secondary clinical endpoints displayed correlation between overall survival (P = 0.037) and event-free survival (P = 0.044) when patients were stratified into tertiles by BIM peptide response. Taken together, these results highlight the potential utility of BH3 profiling in personalized diagnostics of AML by offering actionable information for patient management decisions.


Spatial Patterns and Functional Profiles for Discovering Structure in fMRI Data.
We explore unsupervised, hypothesis-free methods for fMRI analysis in two different types of experiments. First, we employ clustering to identify large-scale functionally homogeneous systems. We formulate a generative mixture model, derive the EM algorithm and apply it to delineate functional systems. We also investigate spectral clustering in application to this problem and demonstrate that both methods give rise to similar partitions of the brain based on resting state fMRI data. Second, we demonstrate how to extend this approach to include information about the experimental protocol. Specifically, we formulate a mixture model in the space of possible profiles of brain response to stimuli. In both applications, our methods confirm previously known results in brain mapping and point to new research directions for exploratory analysis of fMRI data.


Loss of Mitochondrial Localization of Human FANCG Causes Defective FANCJ Helicase.
Fanconi anemia (FA) is a unique DNA damage repair pathway. To date, 22 genes have been identified that are associated with the FA pathway. A defect in any of those genes causes genomic instability, and the patients bearing the mutation become susceptible to cancer. In our earlier work, we identified that Fanconi anemia protein G (FANCG) protects the mitochondria from oxidative stress. In this report, we have identified eight patients having a mutation (C.65G>C), which converts arginine at position 22 to proline (p.Arg22Pro) in the N terminus of FANCG. The mutant protein, hFANCGR22P, is able to repair the DNA and able to retain the monoubiquitination of FANCD2 in the FANCGR22P/FGR22P cell. However, it lost mitochondrial localization and failed to protect mitochondria from oxidative stress. Mitochondrial instability in the FANCGR22P cell causes the transcriptional downregulation of mitochondrial iron-sulfur cluster biogenesis protein frataxin (FXN) and the resulting iron deficiency of FA protein FANCJ, an iron-sulfur-containing helicase involved in DNA repair.


Effects of cardiac growth on electrical dyssynchrony in the single ventricle patient.
Single ventricle patients, including those with hypoplastic left heart syndrome (HLHS), typically undergo three palliative heart surgeries culminating in the Fontan procedure. HLHS is associated with high rates of morbidity and mortality, and many patients develop arrhythmias, electrical dyssynchrony, and eventually ventricular failure. However, the correlation between ventricular enlargement and electrical dysfunction in HLHS physiology remains poorly understood. Here we characterize the relationship between growth and electrophysiology in HLHS using computational modeling. We integrate a personalized finite element model, a volumetric growth model, and a personalized electrophysiology model to perform controlled in silico experiments. We show that right ventricle enlargement negatively affects QRS duration and interventricular dyssynchrony. Conversely, left ventricle enlargement can partially compensate for this dyssynchrony. These findings have potential implications on our understanding of the origins of electrical dyssynchrony and, ultimately, the treatment of HLHS patients.


A systematic review of the role of renin angiotensin aldosterone system genes in diabetes mellitus, diabetic retinopathy and diabetic neuropathy.
The renin angiotensin aldosterone system (RAAS) plays a vital role in regulating glucose metabolism and blood pressure, electrolyte and fluid homeostasis. The aim of this systematic review is to assess the association of the RAAS genes with diabetes mellitus (DM) and its complications of retinopathy, neuropathy and cardiovascular disease (CVD). The relevant English-language studies were identified using the key words of DM, type 1 diabetes mellitus (T1DM), T2DM, renin angiotensin aldosterone polymorphisms or genotypes and RAAS from the search engines of MEDLINE/PubMed, and Scopus from January 1, 1995 to July 30, 2014. Inclusion criteria for selecting relevant studies were reporting the role of RAAS gene variants in the pathogenesis of T1DM or T2DM, diabetic retinopathy (DR), diabetic neuropathy and cardiovascular complication of DM. The reviewers identified 204 studies of which 73 were eligible for inclusion in the present systematic review. The review indicates the angiotensinogen (AGT) M235T polymorphism might not affect the risk of DM. The role of angiotensin converting enzyme insertion/deletion (ACE I/D) and angiotensin II type 1 receptor gene (AT1R) A1166C polymorphisms in the pathogenesis of DM could not be established. Studies indicate the absence of an association between three polymorphisms of AGT M235T, ACE I/D and AT1R A1166C and DR in DM patients. A protective role for ACE II genotype against diabetic peripheral neuropathy has been suggested. Also, the ACE I/D polymorphism might be associated with the risk of CVD in DM patients. More studies with adequate sample size that investigate the influence of all RAAS gene variants together on the risk of DM and its complications are necessary to provide a more clear picture of the RAAS genes polymorphisms involvement in the pathogenesis of DM and its complications.


Cerebral air embolism from a Central Venous Catheter: A timely reminder of the importance of rapid diagnosis.
Cerebral air embolism (CAE) is a rare, avoidable and potentially fatal iatrogenic complication. Here, we report a case of CAE associated with a central venous catheter in the internal jugular vein that resulted in neurological deficits and generalised epileptic seizures. A 64-year-old man admitted for fasciotomy for compartment syndrome developed CAE with left-sided neurological deficits. The suspected origin was retrograde air flow from the right internal jugular venous catheter. The air spontaneously resorbed without the need for specific therapy, and he made a good recovery. CAE is an infrequent iatrogenic complication that requires prompt diagnosis to avoid significant morbidity and mortality. This case serves as a timely reminder that adverse outcome such as stroke, seizures or death can be avoided by a high index of suspicion and prompt diagnosis. Hyperbaric oxygen is the prime therapeutic measure, but high-quality evidence on its clinical value is lacking.


Identification and characterization of outer membrane vesicles from the fish pathogen Vibrio ordalii.
Vibriosis outbreaks due to Vibrio ordalii occur globally, but Chilean salmon aquaculture, in particular, has suffered significant monetary losses in the last 15 years. Little is known about the virulence mechanisms employed by V. ordalii. However, most Vibrio pathogens (e.g., Vibrio anguillarum, a very close taxonomic species) present outer membrane vesicles (OMVs) that are released extracellularly and implicated in the delivery of virulence factors to host cells. This study provides the first reported evidence of the fish pathogen V. ordalii producing and releasing OMVs under normal growth conditions. Analyses were conducted with the V. ordalii strain Vo-LM-18 and the type strain ATCC 33509T . For comparative purposes, the reference strain V. anguillarum ATCC 43307 was employed. The average size for the three Vibrio strains was 0.215 ± 0.6 µm (via scanning electron microscopy) or between 0.19 and 1.8 µm (via dynamic light scattering), with each bacterium presenting a wide range. SDS-PAGE revealed similarities in OMV patterns, but neither total nor external proteins were identical. Comparing V. ordalii ATCC 33509T and Vo-LM-18, bands were most evident in the total proteins, and the greatest degree of similarity in OMV profiles was between 37 and 50 kDa. The purified OMVs demonstrated haemolytic enzyme activity, which could play a role during V. ordalii infection. These data represent an initial step towards gaining new insights into this virulence factor, of which a lot is known in other pathogenic microorganisms.


Specific inhibition of p97/VCP ATPase and kinetic analysis demonstrate interaction between D1 and D2 ATPase domains.
The p97 AAA (ATPase associated with diverse cellular activities), also called VCP (valosin-containing protein), is an important therapeutic target for cancer and neurodegenerative diseases. p97 forms a hexamer composed of two AAA domains (D1 and D2) that form two stacked rings and an N-terminal domain that binds numerous cofactor proteins. The interplay between the three domains in p97 is complex, and a deeper biochemical understanding is needed in order to design selective p97 inhibitors as therapeutic agents. It is clear that the D2 ATPase domain hydrolyzes ATP in vitro, but whether D1 contributes to ATPase activity is controversial. Here, we use Walker A and B mutants to demonstrate that D1 is capable of hydrolyzing ATP and show for the first time that nucleotide binding in the D2 domain increases the catalytic efficiency (kcat/Km) of D1 ATP hydrolysis 280-fold, by increasing kcat 7-fold and decreasing Km about 40-fold. We further show that an ND1 construct lacking D2 but including the linker between D1 and D2 is catalytically active, resolving a conflict in the literature. Applying enzymatic observations to small-molecule inhibitors, we show that four p97 inhibitors (DBeQ, ML240, ML241, and NMS-873) have differential responses to Walker A and B mutations, to disease-causing IBMPFD mutations, and to the presence of the N domain binding cofactor protein p47. These differential effects provide the first evidence that p97 cofactors and disease mutations can alter p97 inhibitor potency and suggest the possibility of developing context-dependent inhibitors of p97.


Electronic criterion for categorizing the chalcogen and halogen bonds: sulfur-iodine interactions in crystals.
Diversity of mutual orientations of Y-S and I-X and covalent bonds in molecular crystals complicate categorizing noncovalent chalcogen and halogen bonds. Here, the different types of S...I interactions with short interatomic distances are analysed. The selection of S...I interactions for the categorization of the chalcogen and halogen bonds has been made using angles that determine the mutual orientation of electron lone pairs and σ-holes interacted S and I atoms. In complicated cases of noncovalent interactions with `hole-to-hole' of S and I orientations, distinguishing the chalcogen and halogen bonds is only possible if the atom is uniquely determined, which also provides the electrophilic site. The electronic criterion for chalcogen/halogen bonds categorizing that is based on analysis of dispositions of electron density and electrostatic potential minima along the interatomic lines has been suggested and its effectiveness has been demonstrated.


Serum beta-carotene and vitamin C as biomarkers of vegetable and fruit intakes in a community-based sample of French adults.
Relatively high intakes of vegetables and fruit and relatively low intakes of fat are associated with lower rates of heart disease and many types of cancer. Biomarkers for vegetable and fruit consumption are most useful when applicable across different ages, body weights, diets, and varying patterns of fat intake. This study examined two biomarkers, serum concentrations of beta-carotene and vitamin C, as a function of anthropometric, dietary, and lifestyle factors in a community-based sample of French adults. The interview-based dietary-history method was used to assess dietary intakes of 361 males and 476 females aged 18-94 y resident in the Val-de-Marne district southeast of Paris. Serum beta-carotene was quantified by HPLC and vitamin C was measured by using an automated method. Serum beta-carotene and vitamin C concentrations were positively associated with vegetable and fruit intakes and were negatively linked to the consumption of energy, alcohol, and fat. Multiple-regression analyses showed that serum beta-carotene concentration was predicted by fruit and vegetable intakes but was inversely associated with body mass, energy and alcohol intakes, and tobacco use. Serum vitamin C concentration was positively associated with fruit consumption but was negatively associated with age, body mass, and tobacco use. Serum beta-carotene and vitamin C concentrations are useful biomarkers of vegetable and fruit consumption in the French diet. However, other dietary and lifestyle factors also have a significant effect on circulating concentrations of these antioxidant micronutrients.


[Prognostic significance of the hepatopulmonary syndrome in liver cirrhosis].
The prognosis of hepatopulmonary syndrome (HPS) has been only rarely investigated. We investigated the survival of 32 cirrhotic patients, 14 (44%) with HPS and 18 with a normal gaseous exchange (NGE), and the associated factors. During a mean (standard deviation) of 56 (27) months, 9 patients in the HPS group (relative risk: 0.64) and 4 patients in the NGE group (relative risk: 0.22) died. The odds ratio was 6.42 (p < 0.01; 95% confidence interval, 0.04-0.80). Patients in the HPS group died after 44 (31) months, while patients in the NGE group died 65 (21) months after inclusion (p < 0.05). Overall, 46% of deaths were liver related. Factors associated with death were the right to left shunting and the increased plasmatic renin levels. Of note, the plasmatic volume and diffusing capacity were protective. The coexistence of HPS worsens the prognosis in liver cirrhosis.


[Efficiency of outpatient clinics in Matanzas, Cuba, using data envelopment analysis].
To rate the efficiency of all the outpatient clinics in Matanzas, Cuba; identify the best-performing clinics; and find opportunities for improvement at the others. A descriptive study of the 40 outpatient clinics in the province of Matanza was carried out during the first trimester of 2006. Clinics were grouped according to the complexity of services they offer and the socioeconomic level of the municipality in which they are located. Five output and six input variables were analyzed. Calculations were performed using data envelopment analysis, including optimization of results and constant and variable returns-to-scale. In general, the clinics studied had high efficiency rates, with a mean of 0.95 +/- 0.11. Eleven (27.5%) clinics studied were rated inefficient (0.77 +/- 0.12). The following Three key areas for improvement were identified: increasing tuberculosis detection rates in the community, reducing rates of preventable infant mortality, and expanding immunization coverage. Among the 11 clinics rated as inefficient, there were resource gaps in at least one indicator. By recognizing successful clinics, best practices were identified that could be used to improve the weaker clinics. The resource distribution process should be reviewed to ensure that additional, unneeded resources are not used to improve efficiency.


Slow Magnetic Relaxation of a 12-Metallacrown-4 Complex with a Manganese(III)-Copper(II) Heterometallic Ring Motif.
The heterobimetallic metallacrown (MC), (TMA)2{Mn(OAc)2[12-MCMn(III)Cu(II)N(shi)-4](CH3OH)}·2.90CH3OH, 1, where TMA+ is tetramethylammonium, -OAc is acetate, and shi3- is salicylhydroximate, consists of a MnII ion captured in the central cavity and alternating unambiguous and ordered manganese(III) and copper(II) sites about the MC ring, a first for the archetypal MC structure design. DC-magnetometry characterization and subsequent simulation with the Spin Hamiltonian H = -J1(s1 + s3)·s5 - J2(s2 + s4)·s5 - J3Σi=14si·si+1 + d(sz,12 + sz,32) + μBΣj=15gjsj·B indicates an S = 5/2 ground state and a sizable axial zero-field splitting on MnIII. AC-susceptibility measurements reveal that 1 displays slow magnetization relaxation akin to single-molecule magnet (SMM) behavior.


Urinary tract infection mimicking acute mesenteric ischaemia in an immunocompromised patient.
Bowel infarction due to acute mesenteric ischaemia (AMI) is an abdominal emergency with a high mortality rate. We report a case of exaggerated septic response to a urinary tract infection mimicking AMI in an immunosuppressed diabetic patient. A 56-year-old female was found collapsed at home with a 24 hour history of diarrhoea, a central abdominal pain and a complex past medical history. Examination showed her to be pyrexial, drowsy, profoundly dehydrated with evidence of cardiovascular collapse. She had a tender distended abdomen, raised inflammatory markers, raised lactate of 9.1 u/L and urinalysis was positive for leucocytes and nitrites. An abdominal computed tomography (CT) scan was reported to show small bowel ischaemia. She underwent a negative laparotomy and recovered following management in the intensive therapy unit. The negative laparotomy rate can be reduced by having abdominal CT performed and reported by an experienced radiologist or by the use of diagnostic laparoscopy.


Experimental fascial flap model in the dog: free flap of the dorsal thoracic fascia.
For years, various types of fascial flaps have been used in clinical practice; however, there are many unanswered questions regarding their basic physiology, anatomy and histopathologic changes occurring after transfer. Simple and reliable flap models are needed to investigate these questions, but very few of these flap models have been described in experimental animals to date. The purpose of this study was to describe a new reliable fascia flap model in the dog-the dorsal thoracic fascia flap. This fascia is defined as the anatomic layer that contains the blood supply to the scapular and parascapular fasciocutaneous flaps. Fourteen adult dogs were used in this experiment. The vascular anatomy of the dorsal thoracic fascia was studied by anatomic dissection and microangiography. Anatomic dissection revealed that the main axial vessel supplying the dorsal thoracic fascia was the superficial branch of the thoracodorsal vessel. Based on the vascular pedicle, fascia flaps generally measuring 15 x 24 cm were created. At gross observation, all of these large flaps based solely on the vascular pedicle were observed to be well-perfused. Microangiographic examination revealed the intense vascularity of the superficial branches of the thoracodorsal vessels in the whole area of all flaps. It was concluded that this is a simple and reliable fascial flap model which can be prepared as a free or pedicled flap. It has a consistent, long vascular pedicle with large vessel diameters supporting a large fascial flap.


Transmission of 'Candidatus Anaplasma camelii' to mice and rabbits by camel-specific keds, Hippobosca camelina.
Anaplasmosis, caused by infection with bacteria of the genus Anaplasma, is an important veterinary and zoonotic disease. Transmission by ticks has been characterized but little is known about non-tick vectors of livestock anaplasmosis. This study investigated the presence of Anaplasma spp. in camels in northern Kenya and whether the hematophagous camel ked, Hippobosca camelina, acts as a vector. Camels (n = 976) and > 10,000 keds were sampled over a three-year study period and the presence of Anaplasma species was determined by PCR-based assays targeting the Anaplasmataceae 16S rRNA gene. Camels were infected by a single species of Anaplasma, 'Candidatus Anaplasma camelii', with infection rates ranging from 63-78% during the dry (September 2017), wet (June-July 2018), and late wet seasons (July-August 2019). 10-29% of camel keds harbored 'Ca. Anaplasma camelii' acquired from infected camels during blood feeding. We determined that Anaplasma-positive camel keds could transmit 'Ca. Anaplasma camelii' to mice and rabbits via blood-feeding. We show competence in pathogen transmission and subsequent infection in mice and rabbits by microscopic observation in blood smears and by PCR. Transmission of 'Ca. Anaplasma camelii' to mice (8-47%) and rabbits (25%) occurred readily after ked bites. Hence, we demonstrate, for the first time, the potential of H. camelina as a vector of anaplasmosis. This key finding provides the rationale for establishing ked control programmes for improvement of livestock and human health.


The StartReact effect on self-initiated movements.
Preparation of the motor system for movement execution involves an increase in excitability of motor pathways. In a reaction time task paradigm, a startling auditory stimulus (SAS) delivered together with the imperative signal (IS) shortens reaction time significantly. In self-generated tasks we considered that an appropriately timed SAS would have similar effects. Eight subjects performed a ballistic wrist extension in two blocks: reaction, in which they responded to a visual IS, and action, in which they moved when they wished within a predetermined time window. In 20-25% of the trials, a SAS was applied. We recorded electromyographic activity of wrist extension and wrist movement kinematic variables. No effects of SAS were observed in action trials when movement was performed before or long after SAS application. However, a cluster of action trials was observed within 200 ms after SAS. These trials showed larger EMG bursts, shorter movement time, shorter time to peak velocity, and higher peak velocity than other action trials (P < 0.001 for all), with no difference from Reaction trials containing SAS. The results show that SAS influences the execution of self-generated human actions as it does with preprogrammed reaction time tasks during the assumed building up of preparatory activity before execution of the willed motor action.


How Effectively Can Adaptive Sampling Methods Capture Spontaneous Ligand Binding?
Molecular dynamics (MD) simulations that capture the spontaneous binding of drugs and other ligands to their target proteins can reveal a great deal of useful information, but most drug-like ligands bind on time scales longer than those accessible to individual MD simulations. Adaptive sampling methods-in which one performs multiple rounds of simulation, with the initial conditions of each round based on the results of previous rounds-offer a promising potential solution to this problem. No comprehensive analysis of the performance gains from adaptive sampling is available for ligand binding, however, particularly for protein-ligand systems typical of those encountered in drug discovery. Moreover, most previous work presupposes knowledge of the ligand's bound pose. Here we outline existing methods for adaptive sampling of the ligand-binding process and introduce several improvements, with a focus on methods that do not require prior knowledge of the binding site or bound pose. We then evaluate these methods by comparing them to traditional, long MD simulations for realistic protein-ligand systems. We find that adaptive sampling simulations typically fail to reach the bound pose more efficiently than traditional MD. However, adaptive sampling identifies multiple potential binding sites more efficiently than traditional MD and also provides better characterization of binding pathways. We explain these results by showing that protein-ligand binding is an example of an exploration-exploitation dilemma. Existing adaptive sampling methods for ligand binding in the absence of a known bound pose vastly favor the broad exploration of protein-ligand space, sometimes failing to sufficiently exploit intermediate states as they are discovered. We suggest potential avenues for future research to address this shortcoming.


Phase II study of dasatinib in Philadelphia chromosome-negative acute and chronic myeloid diseases, including systemic mastocytosis.
Molecular characterization of Philadelphia chromosome-negative (Ph-) chronic myeloproliferative disorders, such as systemic mastocytosis (SM), has provided a clear rationale for investigating novel targeted therapies. The tyrosine kinase (TK) inhibitor dasatinib is 325-fold more potent against Bcr-Abl TK than imatinib in vitro, significantly inhibiting wild-type KIT and platelet-derived growth factor receptor beta TKs, and is active against cells carrying the mutant KIT-D816V gene. In this phase 2, open-label study, the efficacy of dasatinib (140 mg/d) was investigated in 67 patients with various Ph- myeloid disorders, including SM (n = 33; 28 KIT-D816V positive). The overall response rate to dasatinib in patients with SM was 33%. Only two patients, one with SM-myelofibrosis and one with SM-chronic eosinophilic leukemia, achieved complete response (elimination of mastocytosis) lasting for 5 and 16 months, respectively. Both patients were negative for KIT-D816V mutation, had low tryptase levels, abnormal WBC counts, and anemia, and had failed prior therapy with erythropoietin. Additional nine SM patients had symptomatic response, lasting 3 to 18+ months. Complete responses were achieved in two other patients (acute myeloid leukemia and hypereosinophilic syndrome). No responses were observed among patients with myelodysplastic syndromes and primary myelofibrosis. The majority of adverse events were grade 1/2. These data show that dasatinib therapy may benefit a selected group of SM patients, primarily by improving their symptoms, but it does not eliminate the disease in the patients with KIT-D816V mutation.


Unfocused extracorporeal shock waves induce anabolic effects in osteoporotic rats.
Unfocused extracorporeal shock waves (UESW) have been shown to have an anabolic effect on bone mass. Therefore we investigated the effects of UESW on bone in osteoporotic rats with and without anti-resorptive treatment. Twenty-week-old rats were ovariectomized (n = 27). One group was treated with saline and another group with Alendronate (ALN) 2.4 µg/kg, 3×/week. UESW were applied 2 weeks after ovariectomy. Thousand UESW were applied to one hind leg, the contra-lateral hind leg was not treated and served as control. With the use of in vivo micro-CT scanning it was shown that in saline treated rats trabecular bone volume fraction (BV/TV) was higher at 2 weeks follow-up in UESW treated legs compared to control legs. However, at 4 and 10 weeks no difference was found. In ALN treated animals UESW led to a pronounced anabolic response resulting in an increase in BV/TV at all time-points. Furthermore, UESW resulted in increased cortical volume (CtV), higher trabecular connectivity and, more plate-like and thicker trabeculae. Biomechanical testing showed that UESW lead to a higher maximum force before failure and higher stiffness in all treatment groups. With histology abundant areas of intramembranous bone formation along the periosteal cortex and within the bone marrow were observed. In conclusion this study shows promising results for the use of UESW in the treatment of osteoporosis, especially when this treatment is combined with an anti-resorptive treatment.


Spatially invariant image sequences.
The authors define linearly additive spatially invariant image sequences and present an explicit mathematical model for describing them. In such a sequence, all objects are positionally invariant in each image of the sequence but have varying gray-scale contributions to the successive images of the sequence. The various components (features or processes) of the scene or object contribute additively to each image of the sequence, but each component has a characteristic variation (signature) from image to image due to the variation of the function, parameter or spectral band over the sequence. Objects with different spectral characteristics will have different image sequence signatures which can be used to distinguish them. Also presented are the general formulation, derivation, and explicit expression for the linear filter, called the simultaneous diagonalization (SD) filter, that calculates a single new image from the sequence such that a desired process is emphasized and any number of undesired processes is suppressed in the filtered image.


Unusual variants of melanoma: fact or fiction?
Evidence is reviewed separating unusual variants of melanoma from the large group of superficial spreading and nodular (SSM/NOD) histogenetic types. These include (1) the relationship of moles to melanoma of the SSM/ NOD types not found in melanoma arising in lentigo maligna (LMM), desmoplastic neurotrophic melanoma (DNM), mucosal lentiginous melanoma (MLM), or acral lentiginous melanoma (ALM); (2) the strong sunlight association in lentigo maligna (LM) and LMM not always present in SSM/NOD and not likely at all in acral or mucosal lesions (ALM, MLM); (3) epidemiological differences of age, race, and prognosis among the various subtypes; and (4) analogies to neoplasms in other organ systems. These data justify the following conclusions: (1) Variants of melanoma exist as in other neoplasms. (2) They are of epidemiological and therapeutic importance. (3) Until further data are available or networked, data base analysis should use microstage measurements in the common forms of SSM and NOD only, and approach the unusual variants separately and cautiously.