Capacity planning for maternal-fetal medicine using discrete event simulation.
Maternal-fetal medicine is a rapidly growing field requiring collaboration from many subspecialties. We provide an evidence-based estimate of capacity needs for our clinic, as well as demonstrate how simulation can aid in capacity planning in similar environments. A Discrete Event Simulation of the Center for Fetal Diagnosis and Treatment and Special Delivery Unit at The Children's Hospital of Philadelphia was designed and validated. This model was then used to determine the time until demand overwhelms inpatient bed availability under increasing capacity. No significant deviation was found between historical inpatient censuses and simulated censuses for the validation phase (p = 0.889). Prospectively increasing capacity was found to delay time to balk (the inability of the center to provide bed space for a patient in need of admission). With current capacity, the model predicts mean time to balk of 276 days. Adding three beds delays mean time to first balk to 762 days; an additional six beds to 1,335 days. Providing sufficient access is a patient safety issue, and good planning is crucial for targeting infrastructure investments appropriately. Computer-simulated analysis can provide an evidence base for both medical and administrative decision making in a complex clinical environment.


Chemically tunable photoresponse of ultrathin polypyrrole.
Building a complex structure system of conductive polymers without a complicated fabricating process is a long-awaited goal to improving the functional photoresponse properties of conductive polymers. In this study, we demonstrate that the photoresponse of polypyrrole (PPy)-based photodetector devices with an ultrathin polymer layer can be chemically modulated by simply immersing the devices into an alkaline solution. After alkaline treatment, the pyrrole unit transforms into a quinoid structure. Characteristics of current-voltage reveal an increased photosensitivity with several orders of magnitude when decreasing the applied bias voltage. Furthermore, ultrathin PPy belts with a width of 100 nm exhibit ultra-high photosensitivites of roughly 1000 (unit) and photoresponsivities of 54.3 A W-1 due to the high surface area ratio of the nanobelts.


Morphological, functional and neurological outcomes of craniectomy versus cranial vault remodeling for isolated nonsyndromic synostosis of the sagittal suture: a systematic review.
Craniosynostosis is a condition characterized by the premature closure of one or more of the cranial vault sutures. It can occur alone or in association with other congenital defects and may be part of a syndrome. The sagittal suture is most commonly affected, comprising 40-60% of cases. Premature fusion of the sagittal suture can cause scaphocephaly due to compensatory anterior-posterior growth of the skull. This is morphologically considered as a narrow elongated skull with a decreased cephalic index, and is diagnosed clinically and/or radiologically. Both the indications for surgery and the techniques used have varied with time and location. Surgical techniques have evolved, from limited craniectomy to calvarial remodeling. In recent times a return to craniectomy methods has occurred with the more recent introduction of endoscopic methods. The objectives of this review were to identify and synthesize the best available evidence on the morphological, functional and neurological outcomes of craniectomy compared to cranial vault remodeling. This review considered studies of infants with primary isolated sagittal synostosis operated on or before the mean age of 24 months. The intervention of interest was local craniectomy and this was compared to cranial vault remodeling. Morphological (primary), functional and neurological (secondary) outcomes were included. Mortality, complications and aesthetic outcome were included as tertiary outcomes. A comprehensive search was undertaken across major databases. The retrieved studies were assessed by two independent reviewers for methodological validity prior to inclusion. Data was then extracted and, where possible, pooled in statistical meta-analysis. For descriptive studies, where statistical pooling was not possible, the findings are presented in narrative form. Search and retrieval: Based on critical appraisal, 27 studies were considered to be suitable for this review. These studies were all descriptive in nature. Meta-analysis was only possible for the primary morphological outcome (post-operative cephalic index).Morphological (cephalic index):At one year follow-up, post-operatively remodeling offers an advantage over craniectomy (Z = 4.16, P<0.0001)Morphological:Improvements of the cephalic index to varying degrees were seen in patients receiving either procedure and there is not enough evidence to suggest that either treatment group had greater improvement over the other.Functional and neurological:Although their global scores may be comparable to an age-matched population, patients with sagittal synostosis who have undergone a surgical repair of any type may have discrepancies in specific domains and may be at risk of developing learning disorders. There is insufficient primary research with inter-procedure comparisons of preoperative and postoperative cognitive and neurological outcomes.Tertiary outcomes:There is not enough evidence to comment on mortality or postoperative infection in either treatment group. Patients undergoing cranial vault remodeling have a higher rate of transfusion compared to those undergoing craniectomy; however, it is likely that this difference relates to elective transfusion based on hospital-specific protocols. It remains unknown whether there is an inherently higher need for transfusion in patients undergoing remodeling procedures. Delaying surgery however may increase the risk of raised intracranial pressure (ICP) and its associated complications. Whilst there is no evidence for raised ICP post-craniectomy, a few studies have shown raised ICP in patients post-remodeling. There is not enough evidence to establish a relationship between both procedures and raised ICP. Aesthetic outcome appears to be "better" in patients who undergo remodeling; however, there is little rigorous evidence to support this hypothesis. Conclusions were drawn from both the meta-analysis and the narrative results.When comparing the mean change in cephalic index one year after surgery, remodeling was shown to be superior to limited craniectomy in patients with isolated synostosis of the sagittal suture. However both procedures were seen to give improvements at short, medium and longer term time points. Improvements in cephalic index may be sustained, deteriorate or improve over time; based on the current data neither procedure offers a clear long-term advantage over the other. Longer follow-up is required to compare outcomes at different time points.Patients who have surgery (any type) for isolated sagittal synostosis may have deficiencies in different subdomains at later school-age testing, whilst maintaining an age-appropriate global intelligence quotient (IQ) and school performance. There is no evidence to suggest that surgery of either type imparts any benefit in terms of functional or neurological outcomes.There is no evidence to suggest that surgery of either type imparts any benefit in terms of functional or neurological outcomes. While school performance and general IQ may be comparable to age-matched controls, patients with sagittal synostosis who have undergone surgical repair of any type may be at risk of deficiencies in sub-areas of testing and be at risk of learning disorders.There is insufficient evidence regarding mortality, infection, postoperative ICP and aesthetic outcome. While transfusion rates were greater in the remodeling group, this may be due to higher rates of elective transfusion.The inconclusive findings indicate an ongoing need for higher quality primary research comparing the morphological and functional outcomes of craniectomy and cranial vault remodeling in primary sagittal synostosis. Outcomes should be measured in both the short and long term.


Characterization of urban pollutant emission fluxes and ambient concentration distributions using a mobile laboratory with rapid response instrumentation.
A large and increasing fraction of the planet's population lives in megacities, especially in the developing world. These large metropolitan areas generally have very high levels of both gaseous and particulate air pollutants that have severe impacts on human health, ecosystem viability, and climate on local, regional, and even continental scales. Emissions fluxes and ambient pollutant concentration distributions are generally poorly characterized for large urban areas even in developed nations. Much less is known about pollutant sources and concentration patterns in the faster growing megacities of the developing world. New methods of locating and measuring pollutant emission sources and tracking subsequent atmospheric chemical transformations and distributions are required. Measurement modes utilizing an innovative van based mobile laboratory equipped with a suite of fast response instruments to characterize the complex and "nastier" chemistry of the urban boundary layer are described. Instrumentation and measurement strategies are illustrated with examples from the Mexico City and Boston metropolitan areas. It is shown that fleet average exhaust emission ratios of formaldehyde (HCHO), acetaldehyde (CH3CHO) and benzene (C6H6) are substantial in Mexico City, with gasoline powered vehicles emitting higher levels normalized by fuel consumption. NH3 exhaust emissions from newer light duty vehicles in Mexico City exceed levels from similar traffic in Boston. A mobile conditional sampling air sample collection mode designed to collect samples from intercepted emission plumes for later analysis is also described.


Mode-coupling theory for reaction dynamics in liquids.
A theory for chemical reaction dynamics in condensed phase systems based on the generalized Langevin formalism of Grote and Hynes [J. Chem. Phys. 73, 2715 (1980)] is presented. A microscopic approach to calculate the dynamic friction is developed within the framework of a combination of kinetic and mode-coupling theories. The approach provides a powerful analytic tool to study chemical reactions in realistic condensed phase environments. The accuracy of the approach is tested for a model isomerization reaction in a Lennard-Jones fluid. Good agreement is obtained for the transmission coefficient at different solvent densities, in comparison with numerical simulations based on the reactive-flux approach.


Bidirectional mechanobiology between cells and their local extracellular matrix probed by atomic force microscopy.
There is growing recognition that the mechanical interactions between cells and their local extracellular matrix (ECM) are central regulators of tissue development, homeostasis, repair and disease progression. The unique ability of atomic force microscopy (AFM) to probe quantitatively mechanical properties and forces at the nanometer or micrometer scales in all kinds of biological samples has been instrumental in the recent advances in cell and tissue mechanics. In this review we illustrate how AFM has provided important insights on our current understanding of the mechanobiology of cells, ECM and cell-ECM bidirectional interactions, particularly in the context of soft acinar tissues like the mammary gland or pulmonary tissue. AFM measurements have revealed that intrinsic cell micromechanics is cell-type specific, and have underscored the prominent role of β1 integrin/FAK(Y397) signaling and the actomyosin cytoskeleton in the mechanoresponses of both parenchymal and stromal cells. Moreover AFM has unveiled that the micromechanics of the ECM obtained by tissue decellularization is unique for each anatomical compartment, which may support both its specific function and cell differentiation. AFM has also enabled identifying critical mechanoregulatory proteins involved in branching morphogenesis (MMP14) and acinar differentiation (α3β1 integrin), and has clarified the role of altered tissue mechanics and architecture in a variety of pathologic conditions. Critical technical issues of AFM mechanical measurements like tip geometry effects are also discussed.


Absolute Minimal Sampling of Homonuclear 2D NMR TOCSY Spectra for High-Throughput Applications of Complex Mixtures.
Modern applications of 2D NMR spectroscopy to diagnostic screening, metabolomics, quality control, and other high-throughput applications are often limited by the time-consuming sampling requirements along the indirect time domain t1 . 2D total correlation spectroscopy (TOCSY) provides unique spin connectivity information for the analysis of a large number of compounds in complex mixtures, but standard methods typically require >100 t1 increments for an accurate spectral reconstruction, rendering these experiments ineffective for high-throughput applications. For a complex metabolite mixture it is demonstrated that absolute minimal sampling (AMS), based on direct fitting of resonance frequencies and amplitudes in the time domain, yields an accurate spectral reconstruction of TOCSY spectra using as few as 16 t1 points. This permits the rapid collection of homonuclear 2D NMR experiments at high resolution with measurement times that previously were only the realm of 1D experiments.


Dropout and compliance to physical exercise in menopausal osteopenic women: the European "happy bones" project.
Decline in muscle mass and bone density seem to be two of the most disabling side effects of menopause that negatively affect women's quality of life. Promoting physical activity protocols in the workplace can represent a focal point in the prevention and management of several diseases. The study aims to evaluate the compliance and drop-out of menopausal osteopenic women engaged in combined training performed inside and outside the workplace. Strength and balance were analyzed to evaluate the effect of this protocol on osteoporosis prevention and the risk of falling. 73 menopausal women were enrolled in 5 European countries. They performed 72 lessons of a combined training proposed in the working place (IW) or sport center (SC). Out of the total 39 women enrolled in the IW, 12.8% had to leave the program, while out of the 34 women enrolled in SC, 41.2% did not complete the training. According to the compliance results, 47% of women that completed the trained IW and 85% in the SC recorded high compliance (p = 0.019). Moreover, the strength of the lower limbs (p < 0.001) and static balance (p = 0.001) significantly improved in the whole group. In conclusion, proposing well-structured training in the workplace for menopausal women seems to reduce drop-out. Strength and balance results suggest its positive impact on bone health and risk of falls, despite where it is performed.


In vitro activity of BU-2313B against anaerobic bacteria.
The antimicrobial efficacy of BU-2313B, a new antibiotic, was evaluated in comparison with clindamycin, chloramphenicol and cefoxitin against 265 clinical anaerobic isolates. BU-2313B inhibited 88% of 66 isolates of Bacteroides fragilis tested at 2 micrograms/ml, whereas clindamycin, chloramphenicol and cefoxitin inhibited 98, 97 and 92% of them at 2, 8 and 16 micrograms/ml, respectively. BU-2313B was severalfold more effective than chloramphenicol and cefoxitin, but it was eight times less effective than clindamycin.


Inorganic salt-induced phase control and optical characterization of cadmium sulfide nanoparticles.
Phase-controlled synthesis of CdS nanoparticles from zinc-blende to wurtzite has been successfully realized by an inorganic salt-induced process with no use of surfactants or other ligands in an ultrasound-assisted microwave synthesis system. Pure zinc-blende CdS nanoparticles were produced without adding NaCl, while mixed zinc-blende and wurtzite nanoparticles were obtained by adding NaCl/Cd(2+) molar ratios below 1, and pure wurtzite nanoparticles were produced at a molar ratio of 1. The energy bandgap (E(g)) of the CdS nanoparticles calculated from optical absorption spectra increases as the phase transformation from zinc-blende to wurtzite occurs. Additionally, the CdS nanoparticles showed a 489 nm band-edge emission without adding NaCl, and a 501 nm emission when the molar ratios of NaCl to Cd(2+) are 0.25, 0.5 and 1. It was found that the phase transformation originates from the effect of the halide ion Cl(-). We also found that some other halide ions such as Br(-) and I(-) can induce the phase transformation. It is shown that the phase, size and optical properties of the anisotropic nanoparticles can be well tuned by varying the concentration of the halide ions.


The effect of cimetidine on the relative cardioselectivity of atenolol and metoprolol in asthmatic patients.
The effects of chronic cimetidine therapy on the pharmacodynamic properties of atenolol and metoprolol were assessed in eight asthmatic patients using a placebo-controlled trial. When atenolol and metoprolol were administered at doses which achieved an equivalent degree of beta1-adrenoceptor blockade, metoprolol caused a significantly greater reduction in mean forced expiratory volume in one second (FEV1). This demonstrates the greater cardioselectivity of atenolol. The reductions in mean FEV1 induced by either atenolol or metoprolol were each unaffected by cimetidine. Plasma levels of each of the beta-adrenoceptor blockers were not affected by the addition of cimetidine. The FEV1 in one patient was further reduced when cimetidine was administered with metoprolol, although there was no corresponding change in plasma levels of drug. In this patient there was no such further reduction in FEV1 with atenolol. This study has not provided evidence for a pharmacodynamic interaction between cimetidine and either atenolol or metoprolol, but further studies are indicated.


Urinary excretion of beta 2-microglobulin in myeloma patients.
The levels of beta 2-microglobulin in urine and serum were determined in 39 patients with myelomatosis. In 25 patients the serum beta 2-microglobulin was elevated, and in seven of the patients with increased serum beta-microglobulin the urinary excretion of the protein was also increased. It was concluded that the increased urine beta-microglobulin indicates a renal tubular disorder.


Thyroid transcription factor-1 (TTF-1/Nkx2.1/TITF1) gene regulation in the lung.
TTF-1 [thyroid transcription factor-1; also known as Nkx2.1, T/EBP (thyroid-specific-enhancer-binding protein) or TITF1] is a homeodomain-containing transcription factor essential for the morphogenesis and differentiation of the thyroid, lung and ventral forebrain. TTF-1 controls the expression of select genes in the thyroid, lung and the central nervous system. In the lung, TTF-1 controls the expression of surfactant proteins that are essential for lung stability and lung host defence. Human TTF-1 is encoded by a single gene located on chromosome 14 and is organized into two/three exons and one/two introns. Multiple transcription start sites and alternative splicing produce mRNAs with heterogeneity at the 5' end. The 3' end of the TTF-1 mRNA is characterized by a rather long untranslated region. The amino acid sequences of TTF-1 from human, rat, mouse and other species are very similar, indicating a high degree of sequence conservation. TTF-1 promoter activity is maintained by the combinatorial or co-operative actions of HNF-3 [hepatocyte nuclear factor-3; also known as FOXA (forkhead box A)], Sp (specificity protein) 1, Sp3, GATA-6 and HOXB3 (homeobox B3) transcription factors. There is limited information on the regulation of TTF-1 gene expression by hormones, cytokines and other biological agents. Glucocorticoids, cAMP and TGF-beta (transforming growth factor-beta) have stimulatory effects on TTF-1 expression, whereas TNF-alpha (tumour necrosis factor-alpha) and ceramide have inhibitory effects on TTF-1 DNA-binding activity in lung cells. Haplo-insufficiency of TTF-1 in humans causes hypothyroidism, respiratory dysfunction and recurring pulmonary infections, underlining the importance of optimal TTF-1 levels for the maintenance of thyroid and lung function. Recent studies have implicated TTF-1 as a lineage-specific proto-oncogene for lung cancer.


Construction of the co-expression plasmids of fostriecin polyketide synthases and heterologous expression in Streptomyces.
Polyketides are bioactive natural products with diverse bioactivities, and heterologous production of polyketides in easily engineered microbial hosts is preferred for the production of structurally diverse and the therapeutically active polyketides. In this study, heterologous expression of the biosynthetic genes encoding type I polyketide synthases (PKS) involved in biosynthesis of fostriecin, a unique phosphate monoester polyketide antibiotic, was attempted. Fostriecin PKS (Fos-PKS) biosynthetic gene cluster in a total of 48.4 kb were cloned downstream of the act I promoter in two compatible Streptomyces vectors using Red/ET recombination. The co-expression plasmids were sequentially transferred into Streptomyces lividans and Streptomyces coelicolor. Active transcription of the polyketide genes was confirmed by reverse transcription PCR (RT-PCR) analysis, and the metabolites were detected using high-performance liquid chromatography (HPLC). The recombinant strains S. lividans TK24/p6-fosAB-p4-fosCDEF and S. coelicolor M512/p6-fosAB-p4-fosCDEF were obtained for heterologous expression in Streptomyces. Pigmentation was observed in the recombinant strains, whereas the control strain with empty vector displayed no change in pigment production. Active transcription of the polyketide genes was confirmed by RT-PCR analysis and subsequent sequencing. The present study is the first attempt to overexpress Fos-PKS biosynthetic gene cluster in Streptomyces. More studies on heterologous expression of the fostriecin biosynthetic gene cluster would be beneficial for further understanding the mechanisms of its structural as well as the potential pharmaceutically effect.


Intensive Care Unit Structure Variation and Implications for Early Mobilization Practices. An International Survey.
Early mobilization (EM) improves outcomes for mechanically ventilated patients. Variation in structure and organizational characteristics may affect implementation of EM practices. We queried intensive care unit (ICU) environment and standardized ICU practices to evaluate organizational characteristics that enable EM practice. We recruited 151 ICUs in France, 150 in Germany, 150 in the United Kingdom, and 500 in the United States by telephone. Survey domains included respondent characteristics, hospital and ICU characteristics, and ICU practices and protocols. We surveyed 1,484 ICU leaders and received a 64% response rate (951 ICUs). Eighty-eight percent of respondents were in nursing leadership roles; the remainder were physiotherapists. Surveyed ICUs were predominantly mixed medical-surgical units (67%), and 27% were medical ICUs. ICU staffing models differed significantly (P < 0.001 each) by country for high-intensity staffing, nurse/patient ratios, and dedicated physiotherapists. ICU practices differed by country, with EM practices present in 40% of French ICUs, 59% of German ICUs, 52% of U.K. ICUs, and 45% of U.S. ICUs. Formal written EM protocols were present in 24%, 30%, 20%, and 30%, respectively, of those countries' ICUs. In multivariate analysis, EM practice was associated with multidisciplinary rounds (odds ratio [OR], 1.77; P = 0.001), setting daily goals for patients (OR, 1.62; P = 0.02), presence of a dedicated physiotherapist (OR, 2.48; P < 0.001), and the ICU's being located in Germany (reference, United States; OR, 2.84; P < 0.001). EM practice was also associated with higher nurse staffing levels (1:1 nurse/patient ratio as a reference; 1:2 nurse/patient ratio OR, 0.59; P = 0.05; 1:3 nurse/patient ratio OR, 0.33; P = 0.005; 1:4 or less nurse/patient ratio OR, 0.37; P = 0.005). Those responding rarely cited ambulation of mechanically ventilated patients, use of a bedside cycle, or neuromuscular electrical stimulation as part of their EM practice. Physical therapy initiation, barriers to EM practice, and EM equipment were highly variable among respondents. International ICU structure and practice is quite heterogeneous, and several factors (multidisciplinary rounds, setting daily goals for patients, presence of a dedicated physiotherapist, country, and nurse/patient staffing ratio) are significantly associated with the practice of EM. Practice and barriers may be far different based upon staffing structure. To achieve successful implementation, whether through trials or quality improvement, ICU staffing and practice patterns must be taken into account.


Perceived child weight status, family structure and functioning, and support for health behaviors in a sample of bariatric surgery patients.
The purpose of this study is to describe the associations between bariatric surgery patients' perspectives of their child's weight status, family support for eating and exercise behavior change, and family structure and functioning. A cross-sectional descriptive design with pre- and postsurgery (N = 224) patients was used. Demographics, perceptions of child weight status, family support for eating habits and exercise, and family functioning were assessed from patients at a University Bariatric Clinic. Patients who perceived their child to be overweight/obese reported more impaired family functioning, less family exercise participation, and more discouragement for eating habit change in the family compared to patients who did not perceive their child to be overweight/obese. Single parents more often perceived their children to be overweight/obese, and had more impaired family functioning, and less support for changing eating habits and family exercise participation. Patients with impaired family functioning reported less support for changing eating habits and family exercise participation. Bariatric patients who perceived their child to be overweight/obese and identified as single parents reported more impaired family functioning and less support for eating habits and family participation in exercise. Assessing pre- and postsurgery measures from parents and children will allow the further identification of relationship variables that can be targeted to promote positive family changes that benefit parents and children long-term. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Schistosomiasis vaccines: Farewell to the God of Plague?
Control and even eradication of schistosomiasis have been achieved in some countries using integrated measures but this disease remains endemic in 74 countries with 600 million at risk of whom 200 million are currently infected. The application of control measures, particularly population-based chemotherapy, has in many countries greatly reduced the incidence of serious disease manifestations, but vaccines are urgently needed to supplement existing control measures. Great advances are being made in vaccine development and the first clinical trials are expected to begin shortly.


Determining standards for professional divers diving in benzene polluted waters.
The main objective of this study is to calculate the hours of diving in benzene-polluted waters acceptable for professional divers. We considered recommended absorption limit as that from pulmonary workplace absorption during an 8-h workday at recommended exposure limits set by the National Institute of Occupational Safety and Health, and developed a formula to determine recommended limits for diving time based on actual water and sediment concentrations and exposure conditions. The recommended absorption limit is 1.6 mg of benzene per workday. This is equivalent to total body dermal absorption over a 1-h dive at water concentration of 7.6 mg/L, or absorption from drinking 0.1 L of water with a concentration of 16.5 mg/L, or dermal absorption with 50% of the body covered in sediment with a concentration of 18,851 mg/kg. A formula that calculates allowable diving time considering benzene water and sediment concentrations, and gastrointestinal and dermal exposures is presented. Water concentrations and not exposure to sediment limit recommended diving time, and unlimited diving in areas with sediment and water concentrations thought to pose a serious human health hazard would not surpass the recommended amount of benzene absorbed in the workplace. We conclude that allowable diving time in polluted waters can be calculated resulting in absorption equivalent to recommended limits for pulmonary absorption in the workplace. Our results suggest that agencies determining significant health risk levels of chemical concentrations in sediment and water should consider our findings.


Uniform scale-independent gene transfer to striated muscle after transvenular extravasation of vector.
The muscular dystrophies exemplify a class of systemic disorders for which widespread protein replacement in situ is essential for treatment of the underlying genetic disorder. Somatic gene therapy will require efficient, scale-independent transport of DNA-containing macromolecular complexes too large to cross the continuous endothelia under physiological conditions. Previous studies in large-animal models have revealed a trade-off between the efficiency of gene transfer and the inherent safety of the required surgical and pharmacological interventions to achieve this. Rats and dogs underwent limb or hemibody isolation via atraumatic tourniquet placement or myocardial isolation via heterotopic transplantation. Recombinant adenovirus (10(13) particles per kilogram) or recombinant adeno-associated virus (10(14) genome copies/kg) encoding the lacZ transgene was delivered through pressurized venous infusion without pharmacological mediators. Muscle exhibited almost 100% myofiber transduction in rats and dogs by X-galactosidase staining and significantly higher beta-galactosidase levels compared with nonpressurized delivery. No significant difference was seen in beta-galactosidase levels between 100- or 400-mm Hg groups. The <50-mm Hg group yielded inhomogeneous and significantly lower transgene expression. Uniform scale- and vector-independent skeletal and cardiac myofiber transduction is facilitated by pressurized venous infusion in anatomic domains isolated from the central circulation without pharmacological interference with cardiovascular homeostasis. We provide the first demonstration of uniform gene transfer to muscle fibers of an entire extremity in the dog, providing a firm foundation for further translational studies of efficacy in canine models for human diseases.


[Thymectomy in myasthenia gravis].
Myasthenia gravis is an autoimmundisease characterized by muscle fatigability due to a reduction in available acetylcholinereceptors at neuromuscular junction. Although the role of the thymus in the pathophysiology remains obscure, the results of thymectomy are out of discussion. However, controversy continues concerning the indications for thymectomy. In a period of six years and a half, 24 patients underwent a thymectomy. 22 of the patients benefited from the procedure. Their was no relation with sex, age, and severity of the disease; but how sooner we operate, how greater the changes for a total remission. With this results we propose thymectomy as essential in the treatment of myasthenia gravis. We found no relationship between anatomopathological results and these of the thymectomy. But we can confirm that a reduction in the acetylcholine antibodies titer is not essential for clinical benefit.


Monanema martini in its murid hosts: microfiladermia related to infective larvae and adult filariae.
The microfiladermia of Monanema martini was studied in two natural murid hosts, Lemniscomys striatus and Arvicanthis niloticus, with 137 and 39 rodents respectively inoculated once, twice or several times. Microfilarial densities (mf/mm2) were measured at the ear pinna every three months. Almost all the rodents developed a microfiladermia. When L. striatus rodents were inoculated once with 30, 80, or 400 infective larvae, microfiladermia increased (peaks of 108, 148, 174 mf/mm2 respectively, at six to nine months p.i.); this fits with the fact that, in this filaria-host pair, the number of adult filariae is proportional to the number of inoculated larvae, and remains at a constant level for more than eight months. Nevertheless microfiladermia was limited, especially during the peak, showing the complexity of its regulatory mechanisms. Several low doses over one year, resulting in 145 L3, increased the microfiladermia at the same level than one dose of 400 larvae; the recovery rate of the larvae was reduced but the total number of filariae recovered was increased. A. niloticus, from which the filarial strain originates, showed a much lower microfiladermia than L. striatus (7 mf/mm2 with 80 larvae, at six months p.i.). This was due to a smaller recovery rate of the infective larvae in this host and, overall, to a reduced fertility of the female worms and a shorter lifetime of adult filariae. However, repeated inoculations increased the microfiladermia (32 mf/mm2), due to the constant presence of small numbers of young filariae producing microfilariae. It is to be noted that the two biological systems presented by M. martini in L. striatus and A. niloticus correspond to the two types of ocular pathology described in a recent opthalmological study, chorioretinal atrophy and keratitis respectively.


How to build the non-host plant for stability in insect resistance management.
We consider using non-host plants that express both a toxin and oviposition deterrence for to increase stability for insect resistance management. The two traits reinforce each other ecologically. We used a two-gene model to evaluate this combination of traits. When toxin resistance was recessive or partially recessive, even moderate levels of oviposition deterrence extended time to resistance. When sensitivity to oviposition deterrence started at a low frequency (0.001) selection pressure from the toxin caused the frequency of the gene for sensitivity to oviposition deterrence to increase and the time to resistance was extended beyond the 150-y timeline of the simulations. Even in the worst-case scenario, when toxin resistance was dominant, oviposition deterrence extended time to resistance up to 150 y. The genes for toxin and sensitivity to oviposition deterrence support each other ecologically to prevent resistance from developing to either trait. This creates a more stable insect resistance management strategy.


Enantiomeric recognition of amino acids by amphiphilic crown ethers in Langmuir monolayers.
Four new chiral, amphiphilic crown ethers differing by the hydrophobic tailgroups were synthesized, and their capacity to recognize enantiomeric amino acids was examined using Langmuir films. Surface pressure and surface potential measurements performed on the subphases containing L or D enantiomers of alanine, valine, phenylglycine, and tryptophane indicate that the crown ethers forming the monolayer interact with the amino acids. The effects observed are ascribed to the formation of host-guest complexes. The differences in the magnitude of the effects measured show that the crown ethers are capable of discriminating between different amino acids as well as the enantiomers. Our results demonstrate that the structure of the monolayer plays a decisive role in the molecular recognition process including chiral recognition.


Green Solvent: Green Shadow on Chemical Synthesis.
The natural beauty and purity of our planet has been contaminated deeply due to human selfish activities such as pollution, improper waste management, and various industrial and commercial discharges of untreated toxic by-products into the lap of nature. The collective impact of these hazardous suspensions into the natural habitat is very deadly. Challenges due to human activity on the environment have become ubiquitous. The chemical industry has a major role in human evolution and, predictably, opened gates of increased risk of pollution if the production is not done sustainably. In these circumstances, the notion of Green Chemistry has been identified as the efficient medium of synthesis of chemicals and procedures to eradicate the toxic production of harmful substances. Principles of Green Chemistry guide the scientist in their hunt towards chemical synthesis which requires the use of solvents. These solvents contaminate our air, water, land and surrounding due to its toxic properties. Even though sufficient precautions are taken for proper disposal of these solvents but it is difficult to be recycled. In order to preserve our future and coming generation from the adverse impacts associated with solvents it is very important to find alternative of this which will be easy to use, reusable and also eco-friendly. Solvents are used daily in various industrial processes as reaction medium, as diluters, and in separation procedures. As reaction medium, the role of solvent is to bring catalysts and reactants together and to release heat thus affecting activity and selectivity. The proper selection of the solvent considering its biological, physical and chemical properties is very necessary for product separation, environmental, safety handling and economic factors. Green solvents are the boon in this context. They are not only environmentally benign but also cost effective. The biggest challenge faced by the chemists is adaptation of methods and selection of solvents during chemical synthesis which will give negligible waste product and will remain human and nature friendly. During designing compounds for a particular reaction it is difficult to give assurance regarding the toxicity and biodegradability of the method. Chemists are still far away from predicting the various chemical and biological effects of the compounds on the back of the envelope. To achieve that point is formidable task but it will definitely act as inspiration for the coming generation of chemists. The green solvents are undoubtedly a far better approach to eliminate the negative impacts and aftermath of any chemical synthesis on the environment. Our study in this review covers an overview of green solvents, their role in safer chemical synthesis with reference to some of the important green solvents and their detail summarization.


Draft genome sequence of Streptomyces globisporus C-1027, which produces an antitumor antibiotic consisting of a nine-membered enediyne with a chromoprotein.
Streptomyces globisporus C-1027 is the producer of antitumor antibiotic C-1027, a nine-membered enediyne-containing compound. Here we present a draft genome sequence of S. globisporus C-1027 containing the intact biosynthetic gene cluster for this antibiotic. The genome also carries numerous sets of genes for the biosynthesis of diverse secondary metabolites.


Vitamin D receptor (VDR) expression is not a prognostic factor in cervical cancer.
The aim of this study was to analyze whether VDR status is a prognostic factor that may be of importance for the assessment of recurrence in cervical cancer. VDR-, Ki-67- and p53-status were analyzed immunohistochemically in cervical cancer patients (n=50) and in benign cervical tissue (n=15). The histopathological data of the tumours were evaluated. RNA was extracted from normal cervical tissue (n=4) and cervical carcinomas (n=8) using the method of Chomczynski. RNA was reverse-transcribed and RNA-levels were semiquantitatively detected by PCR. The expression of VDR was significantly increased in cervical cancer compared to normal cervical tissue on the protein-level but not on the RNA-level. No statistically significant correlations were found comparing VDR status with histopathological data (tumour stage, lymph node status, grading, histological tumour type), with the expression of the proliferation marker Ki-67 and of the tumour suppressor gene p53. Our findings indicate that VDR protein expression is not a prognostic factor in cervical cancer. The strong I/DR immunoreactivity that we observed in cervical cancer specimens supports the body of evidence that cervical cancer may be a target for therapeutically applied vitamin D analogues.


Restoration and enhancement of breast ultrasound images using extended complex diffusion based unsharp masking.
Ultrasound is a well-known imaging modality for the interpretation of breast cancer. It is playing very important role for breast cancer detection that are missed by mammograms. The image acquisition is usually affected by the presence of noise, artifacts, and distortion. To overcome such type of issues, there is a need of image restoration and enhancement to improve the quality of image. This paper proposes a single framework for denoising and enhancement of ultrasound images, where a smoothing filter is replaced with an extended complex diffusion-based filter in an unsharp masking technique. The performance evaluation of the proposed method is tested on real ultrasound breast cancer images database and synthetic ultrasound image. The performance evaluation comprises qualitative and quantitative evaluation along with comparative analysis of pre-existing and proposed method. The quantitative evaluation metrics are mean squared error, peak-signal-to-noise ratio, correlation parameter, normalized absolute error, universal quality index, similarity structure index, edge preservation index, a measure of enhancement, a measure of enhancement by entropy, and second derivative like measurement. The result specifies that the proposed method is better suited approach for the removal of speckle noise which follows Rayleigh distribution, restoration of information, enhancement of abnormalities, and proper edge preservation.


Effect of pentoxifylline on motility and membrane integrity of cryopreserved human spermatozoa.
The purpose of this study was to examine the effects of pentoxifylline used before and after semen cryopreservation-thawing on sperm motility and membrane integrity. Twenty-four semen samples were split into four equal aliquots. Aliquots were incubated at 37 degrees C for 30 min, followed by cryopreservation with TEST-yolk freezing medium using slow programmable freezing protocol. After 2 weeks the sperm samples were thawed, washed twice in Quinn's Sperm Washing Medium (modified HTF with 5.0 mg/mL Human Albumin) and incubated at 37 degrees C for 30 min. Aliquots were treated by adding 3 mmol/L pentoxifylline to: (1) fresh sperm samples during incubation period prior to cryopreservation, (2) sperm samples as a supplement to the cryoprotectant prior to cryopreservation, and (3) thawed sperm samples during incubation period. One aliquot received no treatment (control group). The addition of 3 mmol/L pentoxifylline to fresh semen during incubation period prior to cryopreservation significantly decreased progressive and total motility compared with controls. However, the addition of 3 mmol/L pentoxifylline to cryopreserved semen after thawing significantly increased progressive and total motility compared with controls. After post-thaw, no differences in motion characteristics between sperm samples treated by adding 3 mmol/L pentoxifylline as a supplement to the cryoprotectant and control groups were observed. Post-thaw hypoosmotic swelling (HOS) test scores did not improve with the addition of pentoxifylline compared with the control group. It is concluded that pentoxifylline enhanced post-thaw motility of cryopreserved human spermatozoa when added after thawing. No improvement was found by freezing sperm with pentoxifylline.


Design, synthesis and bioactivity study on 5-phenylfuran derivatives as potent reversal agents against P-glycoprotein-mediated multidrug resistance in MCF-7/ADR cell.
P-glycoprotein (P-gp)-mediated multidrug resistance (MDR) is a phenomenon in which cells become resistant to structurally and mechanistically unrelated drugs resulting in low intracellular drug concentrations. It is one of the noteworthy problems in malignant tumor clinical therapeutics. So P-gp protein is one of the ideal targets to solve MDR. Based on the lead compound 5m obtained from our previous work, a series of furan derivatives featuring alkyl-substituted phenols and 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline were designed and synthesized as reversal agents against P-gp in this paper. Compound 16 containing isopropoxy possessed good potency against P-gp mediated MDR in MCF-7/ADR (IC50 (doxorubicin) = 0.73 μM, RF = 69.6 with 5 μM 16 treated). Western blot results and Rh123 accumulation assays showed that 16 effectively inhibited P-gp efflux function but not its expression. The preliminary structure-activity relationship and docking studies demonstrated that compound 16 would be a potential P-gp inhibitor. Most worthy of mention is that compound 16 has achieved satisfactory results in combination with a variety of anti-tumor drugs, such as doxorubicin, paclitaxel, and vincristine. This study forwards a hopeful P-gp inhibitor for withstanding malignant tumor cell with multidrug resistance setting the basis for further studies.


Proguanylin secretion and the role of negative-feedback inhibition in a villous epithelial cell line.
The mechanisms of proguanylin synthesis and secretion in the intestine are incompletely understood. We designed an in vitro model to study proguanylin secretion in a model of intestinal villous epithelial cells. The C2/bbe1 cell line, a differentiated subclone of Caco-2 cells, was used to examine the direction of proguanylin secretion and the potential for feedback regulation via activators of the guanylyl cyclase C signal transduction pathway. When cells were grown on Transwell inserts, proguanylin was secreted into the apical and basolateral media, consistent with other models of intestinal guanylin secretion. Proguanylin synthesis and secretion were not decreased on activation of guanylyl cyclase C-mediated chloride secretion, implying a regulatory system other than negative-feedback inhibition. These data describe the use of C2/bbe1 cells as a model for proguanylin secretion in villous epithelial cells and demonstrate their potential use for the study of the regulatory mechanisms governing proguanylin synthesis and secretion.


Radical contraction of 1,3,2-dioxaphosphepanes to 1,3,2-dioxaphosphorinanes: a kinetic and (17)O NMR spectroscopic study.
Two diastereomeric 5-bromo-4-phenyl-2-phenoxy-2-oxo-1,3,2-dioxophosphepanes have been synthesized and used to study the contraction of 4-phenyl-2-phenoxy-2-oxo-1,3,2-dioxophosphorinan-5-yl radicals. Kinetics were determined by competition methods and demonstrate Arrhenius parameters typical of rearrangements of this kind. Isotopic labeling reveals that all rearrangements are formally of the 1,2-type with retention of configuration at phosphorus. Analysis of the stereochemistry of the rearrangements, however, reveals the two diastereomers to take different paths with respect to the geometry of the presumed alkene radical cation intermediate.


Genetic Diversity and Sequence Polymorphism of Two Genes Encoding Theileria parva Antigens Recognized by CD8+ T Cells among Vaccinated and Unvaccinated Cattle in Malawi.
East Coast fever (ECF) is an acute fatal tick-borne disease of cattle caused by Theileria parva. It causes major losses in exotic and crossbreed cattle, but this could be prevented by a vaccine of T. parva if the vaccine is selected properly based on information from molecular epidemiology studies. The Muguga cocktail (MC) vaccine (Muguga, Kiambu 5 and Serengeti-transformed strains) has been used on exotic and crossbreed cattle. A total of 254 T. parva samples from vaccinated and unvaccinated cattle were used to understand the genetic diversity of T. parva in Malawi using partial sequences of the Tp1 and Tp2 genes encoding T. parva CD8+ antigens, known to be immunodominant and current candidate antigens for a subunit vaccine. Single nucleotide polymorphisms were observed at 14 positions (3.65%) in Tp1 and 156 positions (33.12%) in Tp2, plus short deletions in Tp1, resulting in 6 and 10 amino acid variants in the Tp1 and Tp2 genes, respectively. Most sequences were either identical or similar to T. parva Muguga and Kiambu 5 strains. This may suggest the possible expansion of vaccine components into unvaccinated cattle, or that a very similar genotype already existed in Malawi. This study provides information that support the use of MC to control ECF in Malawi.


The equivalent circuit of single crab muscle fibers as determined by impedance measurements with intracellular electrodes.
The input impedance of muscle fibers of the crab was determined with microelectrodes over the frequency range 1 cps to 10 kc/sec. Care was taken to analyze, reduce, and correct for capacitive artifact. One dimensional cable theory was used to determine the properties of the equivalent circuit of the membrane admittance, and the errors introduced by the neglect of the three dimensional spread of current are discussed. In seven fibers the equivalent circuit of an element of the membrane admittance must contain a DC path and two capacitances, each in series with a resistance. In two fibers, the element of membrane admittance could be described by one capacitance in parallel with a resistance. In several fibers there was evidence for a third very large capacitance. The values of the elements of the equivalent circuit depend on which of several equivalent circuits is chosen. The circuit (with a minimum number of elements) that was considered most reasonably consistent with the anatomy of the fiber has two branches in parallel: one branch having a resistance R(e) in series with a capacitance C(e); the other branch having a resistance R(b) in series with a parallel combination of a resistance R(m) and a capacitance C(m). The average circuit values (seven fibers) for this model, treating the fiber as a cylinder of sarcolemma without infoldings or tubular invaginations, are R(e) = 21 ohm cm(2); C(e) = 47 microf/cm(2); R(b) = 10.2 ohm cm(2); R(m) = 173 ohm cm(2); C(m) = 9.0 microf/cm(2). The relation of this equivalent circuit and another with a nonminimum number of circuit elements to the fine structure of crab muscle is discussed. In the above equivalent circuit R(m) and C(m) are attributed to the sarcolemma; R(e) and C(e), to the sarcotubular system; and R(b), to the amorphous material found around crab fibers. Estimates of actual surface area of the sarcolemma and sarcotubular system permit the average circuit values to be expressed in terms of unit membrane area. The values so expressed are consistent with the dielectric properties of predominantly lipid membranes.


Nanoelectrochemistry at liquid/liquid interfaces for analytical, biological, and material applications.
Herein, we feature our recent efforts toward the development and application of nanoelectrochemistry at liquid/liquid interfaces, which are also known as interfaces between two immiscible electrolyte solutions (ITIES). Nanopipets, nanopores, and nanoemulsions are developed to create the nanoscale ITIES for the quantitative electrochemical measurement of ion transfer, electron transfer, and molecular transport across the interface. The nanoscale ITIES serves as an electrochemical nanosensor to enable the selective detection of various ions and molecules as well as high-resolution chemical imaging based on scanning electrochemical microscopy. The powerful nanoelectroanalytical methods will be useful for biological and material applications as illustrated by in situ studies of solid-state nanopores, nuclear pore complexes, living bacteria, and advanced nanoemulsions. These studies provide unprecedented insights into the chemical reactivity of important biological and material systems even at the single nanostructure level.


[Biopanning of HCV antigen epitopes from a random 12-peptide library by anti-hepatitis C virus polyclonal antibody].
To analyze the epitopes of anti-hepatitis C virus(HCV)antibodies by peptide library biopanning. Phage random peptide library of 12 amino acids was immunoscreened with purified IgG from the sera of hepatitis C patients. Positive clones which were obtained after 3 rounds of biopanning were detected by ELISA and 4 of them were sequenced. After 3 rounds of screening, the radio of output to input increased from (4.6 x 10(-4))% to (5.3 x 10(-2))%, meaning the enrichment was effective. At the third round of screening, all the selected clones proved to specifically react with the sera for immunoscreening. Four positive phage clones were sequenced, which shared a very conservative sequence and was named as C1. Its inserting sequence in the coat protein III was deduced to be GSMSPYVRWYTP, and the positive rate of C1 reacted with 20 cases of HCV patients was 85%. The antigen-mimic peptide C1 is successfully screened from 12 random phage peptide library and it has some diagnostic value.


A prospective, randomized trial examining the efficacy and safety of clarithromycin in combination with ethambutol, rifabutin, or both for the treatment of disseminated Mycobacterium avium complex disease in persons with acquired immunodeficiency syndrome.
This multicenter, randomized, open-label phase 3 clinical trial compared the safety and efficacy of 3 clarithromycin-containing combination regimens for the treatment of disseminated Mycobacterium avium complex (MAC) disease in persons with acquired immunodeficiency syndrome. A total of 160 eligible patients with bacteremic MAC disease were randomized to receive clarithromycin with either ethambutol (C+E), rifabutin (C+R), or both (C+E+R) for 48 weeks. After 12 weeks of treatment, the proportion of subjects with a complete microbiologic response was not statistically significantly different among treatment arms: the proportion was 40% in the C+E group, 42% in the C+R group, and 51% in the C+E+R group (P=.454). The proportion of patients with complete or partial responses who experienced a relapse while receiving C+R (24%) was significantly higher than that of patients receiving C+E+R (6%; P=.027) and marginally higher than that of patients receiving C+E (7%; P=.057). Subjects in the C+E+R group had improved survival, compared with the C+E group (hazard ratio [HR], 0.44; 95% confidence interval [CI], 0.23-0.83) and the C+R group (HR, 0.49; 95% CI, 0.26-0.92).


Regio- and Stereoselective Cascades via Aldol Condensation and 1,3-Dipolar Cycloaddition for Construction of Functional Pyrrolizidine Derivatives.
An efficient and step-economical approach to access functionalized pyrrolizidine derivatives by a one-pot tandem sequence, including an aldol condensation and subsequent 1,3-dipolar cycloaddition process, has been developed, starting from acetone, aldehyde, and proline. A number of substituted aromatic aldehydes were amenable to this transformation, and the desired products, racemic 7a-7w and chiral 9a-9m, were obtained with excellent regioselectivities and outstanding diastereoselectivities. Moreover, in situ NMR studies revealed MgSO4 could effectively promote the aldol condensation pathway in this tandem process.


Alternative approaches to estimation of population pharmacokinetic parameters: comparison with the nonlinear mixed-effect model.
Individual pharmacokinetic parameters can be viewed as independent realizations of a random variable. The probability density function of the variable is assumed to be specified by its first two moments (mean vector and covariance matrix), and these moments then characterize the distribution of the parameters in the population. The following methods are presented for estimation of population characteristics from a set of pharmacokinetic measurements in a sample of subjects: The Global Two-Stage Approach (GTS) uses estimates (and their covariances) of individual parameters obtained after separate fitting of each individual's data. The Iterated Two-Stage Approach (ITS) makes the GTS procedure iterative, using refined bayesian estimates of individual parameters at each step. The Nonlinear Filtering Approach (NLF) also relies on individual parameter estimates produced by using an optimal filter on each subject's data. The three methods give exact results (maximum likelihood estimates), as does NONMEM (the Nonlinear Mixed-Effect Model Approach), when the individual pharmacokinetic model is linear with respect to the parameters and when the distributions of the pharmacokinetic parameters and of the measurement noise in the individual data are both multivariate normal. When the individual pharmacokinetic model is statistically nonlinear (the usual case), the methods differ with respect to: (1) their strategy for handling nonlinearity, (2) their ability to deal with any type of data (experimental and/or routine), and (3) their sensitivity to the amplitude of random effects. With regard to computational aspects, both the computer memory storage requirements and the amount of computation required for the GTS approach are much smaller than for the three other methods. Contrasting considerations as well as results of simulations suggest that GTS, ITS, and, in future, NLF may be valuable alternatives to NONMEM or modifications of it for estimation of population characteristics of pharmacokinetic parameters.


Preliminary Results of Teleconsultations Temporarily Allowed during the COVID-19 Pandemic.
During the COVID-19 pandemic, Korea has temporarily expanded coverage of teleconsultation to ensure access to essential health services. As a preliminary study, we investigated service utilization patterns and the characteristics of doctors and patients involved in these temporary teleconsultation services. Using national health insurance claims data from February 23, 2020 to June 30, 2020 from the Health Insurance Review and Assessment Service, 228269875 cases were identified. Among them, 567390 cases that received teleconsultation services were included in our study. We performed descriptive analyses according to the types of healthcare institutions. In total, 6193 healthcare institutions provided teleconsultation. Of these, 5466 (88.3%) were clinics. Physicians providing teleconsultations were most likely to be doctors of internal medicine (34.0%) or pediatricians (7.0%) and based in the Seoul Metropolitan area (30.4%). In terms of patients undergoing teleconsultation, the most common major disease categories treated were circulatory system diseases (I00-I99). In a detailed analysis, hypertensive diseases (I10-I15) were the most common diagnoses, with a total of 88726 cases (15.6%), followed by diabetes mellitus at 60298 cases (10.6%). The proportion of Medical Aid recipients receiving teleconsultations was higher (9.5%) than other socioeconomic groups. Among all participants, 356622 cases (84.6%) were from a return visit, and 108838 cases (19.2%) received teleconsultation services without being prescribed drugs. Temporarily allowed teleconsultation services were provided mostly to the following patients: 1) those scheduled for revisitation, 2) those with chronic diseases, and 3) those living in pandemic hotspots.


RYR1-Related Myopathies: Clinical, Histopathologic and Genetic Heterogeneity Among 17 Patients from a Portuguese Tertiary Centre.
Pathogenic variants in ryanodine receptor type 1 (RYR1) gene are an important cause of congenital myopathy. The clinical, histopathologic and genetic spectrum is wide. Review a group of the patients diagnosed with ryanodinopathy in a tertiary centre from North Portugal, as an attempt to define some phenotypical patterns that may help guiding future diagnosis. Patients were identified from the database of the reference centre for Neuromuscular Disorders in North Portugal. Their data (clinical, histological and genetic) was retrospectively accessed. Seventeen RYR1-related patients (including 4 familial cases) were identified. They were divided in groups according to three distinctive clinical characteristics: extraocular muscle (EOM) weakness (N = 6), disproportionate axial muscle weakness (N = 2) and joint laxity (N = 5). The fourth phenotype includes patients with mild tetraparesis and no distinctive clinical features (N = 4). Four different histopathological patterns were found: centronuclear (N = 5), central core (N = 4), type 1 fibres predominance (N = 4) and congenital fibre type disproportion (N = 1) myopathies. Each index case, except two patients, had a different RYR1 variant. Four new genetic variants were identified. All centronuclear myopathies were associated with autosomal recessive inheritance and EOM weakness. All central core myopathies were caused by pathogenic variants in hotspot 3 with autosomal dominant inheritance. Three genetic variants were reported to be associated to malignant hyperthermia susceptibility. Distinctive clinical features were recognized as diagnostically relevant: extraocular muscle weakness (and centronuclear pattern on muscle biopsy), severe axial weakness disproportionate to the ambulatory state and mild tetraparesis associated with (proximal) joint laxity. There was a striking genetic heterogeneity, including four new RYR1 variants.


Gouy's Phase Anomaly in Electron Waves Produced by Strong-Field Ionization.
Ionization of atoms by strong laser fields produces photoelectron momentum distributions that exhibit modulations due to the interference of outgoing electron trajectories. For a faithful modeling, it is essential to include previously overlooked phase shifts occurring when trajectories pass through focal points. Such phase shifts are known as Gouy's phase anomaly in optics or as Maslov phases in semiclassical theory. Because of Coulomb focusing in three dimensions, one out of two trajectories in photoelectron holography goes through a focal point as it crosses the symmetry axis in momentum space. In addition, there exist observable Maslov phases already in two dimensions. Clustering algorithms enable us to implement a semiclassical model with the correct preexponential factor that affects both the weight and the phase of each trajectory. We also derive a simple rule to relate two-dimensional and three-dimensional models for linear polarization. It explains the shifted interference fringes and weaker high-energy yield in three dimensions. The results are in excellent agreement with solutions of the time-dependent Schrödinger equation.


Voltage-gated currents in muscle cells of Schistosoma mansoni.
Three morphologically distinct types of muscle fibres isolated from the platyhelminth Schistosoma mansoni have been studied with whole cell current- and voltage-clamp techniques. Fibres showed a marked time-dependent decrease in membrane resistance in response to depolarizing current injections. Voltage-clamp experiments revealed the presence of two distinct voltage-gated outward currents. The most prominent current is a slowly activating, slowly and incompletely inactivating potassium current similar to delayed rectifier currents which have been described in a variety of cell types from a variety of organisms. Also present is a faster activating, quickly and completely inactivating potassium current that shares functional characteristics with 'A'-currents. All three of the cell types studied possess a delayed rectifier current, but only two of the three types have 'A'-currents. Though depolarization with high K+ leads to contraction of the dispersed fibres, no voltage-gated inward currents could be detected by whole cell voltage-clamp under any of our conditions.


Band neutrophil counts are unnecessary for the diagnosis of infection in patients with normal total leukocyte counts.
The authors have assessed the clinical value of an "accurate" band count as a correlate to infection in 33 patients with a culture proven bacterial infection and a normal total leukocyte count. Thirty hematologically normal outpatients, presumably with no infections, were selected for the control group. Manual 500 cell differentials were performed on each subject in both groups. The absolute and percent band count, the absolute and percent total neutrophil count, and the presence of neutrophils more immature than bands were analyzed for their ability to identify the patients with an infection. The presence of neutrophils more immature than bands and the absolute neutrophil count are the two values that best separated the infected patients from the outpatients, even though the selected patients had a normal total leukocyte count. The band count had minimal value for identifying infected patients. Other studies were reviewed regarding the utility of band counts. The authors conclude that manual band counts are unnecessary because when accompanied by a clinical assessment of the patient, automated instruments accurately provide the appropriate hematologic data that may aid in the diagnosis of infection.


Validating the German version of the Quality of Relationship Inventory: confirming the three-factor structure and report of psychometric properties.
Research on psychosocial influences such as relationship characteristics has received increased attention in the clinical as well as social-psychological field. Several studies demonstrated that the quality of relationships, in particular with respect to the perceived support within intimate relationships, profoundly affects individuals' mental and physical health. There is, however, a limited choice of valid and internationally known assessments of relationship quality in Germany. We report the validation of the German version of the Quality of Relationships Inventory (QRI). First, we evaluated its factor structure in a representative German sample of 1.494 participants by means of confirmatory factor analysis. Our findings support the previously proposed three-factor structure. Second, importance and satisfaction with different relationship domains (family/children and relationship/sexuality) were linked with the QRI scales, demonstrating high construct validity. Finally, we report sex and age differences regarding the perceived relationship support, conflict and depth in our German sample. In conclusion, the QRI is a reliable and valid measurement to assess social support in romantic relationships in the German population.


Profiles of telomeric repeats in Insecta reveal diverse forms of telomeric motifs in Hymenopterans.
Telomeres consist of highly conserved simple tandem telomeric repeat motif (TRM): (TTAGG)n in arthropods, (TTAGGG)n in vertebrates, and (TTTAGGG)n in most plants. TRM can be detected from chromosome-level assembly, which typically requires long-read sequencing data. To take advantage of short-read data, we developed an ultra-fast Telomeric Repeats Identification Pipeline and evaluated its performance on 91 species. With proven accuracy, we applied Telomeric Repeats Identification Pipeline in 129 insect species, using 7 Tbp of short-read sequences. We confirmed (TTAGG)n as the TRM in 19 orders, suggesting it is the ancestral form in insects. Systematic profiling in Hymenopterans revealed a diverse range of TRMs, including the canonical 5-bp TTAGG (bees, ants, and basal sawflies), three independent losses of tandem repeat form TRM (Ichneumonoids, hunting wasps, and gall-forming wasps), and most interestingly, a common 8-bp (TTATTGGG)n in Chalcid wasps with two 9-bp variants in the miniature wasp (TTACTTGGG) and fig wasps (TTATTGGGG). Our results identified extraordinary evolutionary fluidity of Hymenopteran TRMs, and rapid evolution of TRM and repeat abundance at all evolutionary scales, providing novel insights into telomere evolution.


Disfluent difficulties are not desirable difficulties: the (lack of) effect of Sans Forgetica on memory.
Scientists working at the intersection of cognitive psychology and education have developed theoretically-grounded methods to help people learn. One important yet counterintuitive finding is that making information harder to learn - that is, creating desirable difficulties - benefits learners. Some studies suggest that simply presenting information in a difficult-to-read font could serve as a desirable difficulty and therefore promote learning. To address this possibility, we examined the extent to which Sans Forgetica, a newly developed font, improves memory performance - as the creators of the font claim. Across four experiments, we set out to replicate unpublished findings by the font's creators. Subjects read information in Sans Forgetica or Arial, and rated how difficult the information was to read (Experiment 1) or attempted to recall the information (Experiments 2-4). Although subjects rated Sans Forgetica as being more difficult to read than Arial, Sans Forgetica led to equivalent memory performance, and sometimes even impaired it. These findings suggest that although Sans Forgetica promotes a feeling of disfluency, it does not create a desirable difficulty or benefit memory.


When Is Evidence Enough Evidence? A Systematic Review and Meta-Analysis of the Trabectome as a Solo Procedure in Patients with Primary Open-Angle Glaucoma.
The purpose of this systematic review and meta-analysis was to examine the availability of evidence for one of the earliest available minimally invasive glaucoma surgery (MIGS) procedures, the Trabectome. Various databases were searched up to December 20, 2016, for any published studies assessing the use of the Trabectome as a solo procedure in patients with primary open-angle glaucoma (POAG). The standardized mean differences (SMD) were calculated for the change in intraocular pressure (IOP) and number of glaucoma mediations used at 1-month, 6-month, and 12-month follow-up. After screening, three studies and one abstract with analyzable data were included. The meta-analysis showed statistically significant reductions in IOP and number of glaucoma medications used at all time points. Though the Trabectome as a solo procedure appears to lower IOP and reduces the number of glaucoma medications, more high-quality studies are required to make definitive conclusions. The difficulty of obtaining evidence may be one of the many obstacles that limit a full understanding of the potential safety and/or efficacy benefits compared to standard treatments. The time has come for a thoughtful and integrated approach with stakeholders to determine optimal access to care strategies for our patients.


Delay of movement caused by disruption of cortical preparatory activity.
We tested the hypothesis that delay-period activity in premotor cortex is essential to movement preparation. During a delayed-reach task, we used subthreshold intracortical microstimulation to disrupt putative "preparatory" activity. Microstimulation led to a highly specific increase in reach reaction time. Effects were largest when activity was disrupted around the time of the go cue. Earlier disruptions, which presumably allowed movement preparation time to recover, had only a weak impact. Furthermore, saccadic reaction time showed little or no increase. Finally, microstimulation of nearby primary motor cortex, even when slightly suprathreshold, had little effect on reach reaction time. These findings provide the first evidence, of a causal and temporally specific nature, that activity in premotor cortex is fundamental to movement preparation. Furthermore, although reaction times were increased, the movements themselves were essentially unperturbed. This supports the suggestion that movement preparation is an active and actively monitored process and that movement can be delayed until inaccuracies are repaired. These results are readily interpreted in the context of the recently developed optimal-subspace hypothesis.


Chronic myofascial pain: management by low-output helium-neon laser therapy.
Therapeutic benefits of low-output helium-neon laser therapy have not been established, but laser therapy has been suggested as an effective means of treating many acute and chronic musculoskeletal pain syndromes. Although not released for general clinical use by the FA, the helium-neon laser has been promoted to physical therapists and athletic trainers as potentially useful for the treatment of pain syndromes. In particular, it has been proposed that it may be more effective than conventional measures such as medication and conventional physical therapy in the treatment of myofascial pain syndromes (fibrositis, fibromyalgia). The citations in the literature include only case reports. Sixty-two patients were treated by using acupuncture points. Two sessions of five treatments were given six weeks apart. A crossover double-blind technique was used in the treatments. The clinical responses were assessed using portions of the McGill Pain Questionnaire. No statistical difference between the treatment and the placebo groups could be determined.


[Medical circles in Shimonoseki in the 18th century and Gyuzan Katsuki's three pupils].
According to the diary of Kyokuso Hirose, there is a grave of Dokushoan Nagadomi not only in Osaka but also in Shimonoseki. Following the notes in the diary, I have been looking for his grave. However, I have not found it yet. While doing this research, I came across the tombstone of a doctor, Genyu Tou. A message of more than 300 words was inscribed on his tombstone. It tells us about the history of medicine in Shimonoseki. Its four main points are as follows: 1. Genyu Tou was a pupil under Gyuzan Katsuki. 2. Yuan Nagadomi studied under the same teacher, and both had their own clinics in Ojicho. 3. The second son of Genyu Tou was Sachu Tou, who contributed a lot to the field of medicine as a doctor in Kyoto. 4. Monan Fujii, who wrote this message, studied at the same medical school. This paper will first consider the message engraved on the tombstone in detail, and will introduce Genyu Tou. Secondly, it will describe the two doctors Monan Fujii and Yuan Nagadomi. Finally, this paper will consider the characteristics of medicine in Shimonoseki in the 18th century, and discuss what happened and how these doctors contributed to the new trends of medicine at that time.


Transdermal delivery of atorvastatin calcium from novel nanovesicular systems using polyethylene glycol fatty acid esters: Ameliorated effect without liver toxicity in poloxamer 407-induced hyperlipidemic rats.
Atorvastatin calcium (ATV), a cholesterol-lowering agent, suffers from poor systemic availability (14%) after oral administration in addition to other side effects on the gastrointestinal tract, liver and muscle. The goal of the present investigation was to improve ATV bioavailability and overcome complications attendant with peroral administration by developing a new nanovesicular system encapsulating ATV for its delivery via the transdermal route. The vesicular systems were prepared by incorporating different polyethylene glycol fatty acid esters such as Labrasol, Cremophor EL, Gelucire 44/14 and Tween 80 as edge activators (EAs) in the lipid bilayer. The effect of the phosphatidylcholine (PC):EA molar ratio on the physicochemical properties of the vesicles was investigated. The pharmacokinetic studies of the optimized formulation were evaluated in rats. The optimized formulation was tested in poloxamer 407-induced hyperlipidemic rats. The plasma lipid profile, activity of liver enzymes, and oxidative stress parameters were measured using commercially available kits. The results revealed high ATV entrapment efficiency (EE%) ranging from 55.62 to 83.91%. The formulations that contained Labrasol showed the highest EE%. The mean diameter of the vesicles was in the range of 186-583nm. T8 containing Gelucire 44/14 as an EA in the molar ratio of 15:1 (PC:EA) gave the smallest size and exhibited the best permeation parameters across the skin. The pharmacokinetic studies revealed that about three times statistically significant (p<0.05) improvement in bioavailability, after transdermal administration of nanotransfersomal ATV gel compared to oral ATV suspension. The transdermal vesicular system exhibited a significant decrease in plasma total cholesterol, triglycerides and LDL cholesterol comparable to oral ATV. Additionally, it lowered the malondialdehyde levels in plasma and abolished the increase in liver enzyme activity. The results obtained suggest that the proposed transdermal vesicular system can serve as a promising alternative means for delivery of ATV.


Design example for the use of hybrid optical elements in the infrared.

The Petzval objective is an effective design form for IR camera systems. It has excellent performance characteristics for high-aperture applications. Its field coverage is limited to a few degrees by the uncorrected Petzval sum.

Using a hybrid aspheric/diffractive front element, it is possible to achieve broadband color correction over the 3-5 µm or 8-12 µm region using only one lens material. For the long wavelength region, Germanium is typically used (except when the application environment is expected to exceed 60° C where Ge becomes opaque). This hybrid combination allows for the correction of the spherical and chromatic aberrations.

The advantage of the high precision obtainable with diamond turning makes it possible to generate simultaneously the diffractive phase profile superimposed on the aspheric surface. To protect the diffractive profile from the environment, it is placed on the rear surface of the first element.


Biomechanics-driven chondrogenesis: from embryo to adult.
Biomechanics plays a pivotal role in articular cartilage development, pathophysiology, and regeneration. During embryogenesis and cartilage maturation, mechanical stimuli promote chondrogenesis and limb formation. Mechanical loading, which has been characterized using computer modeling and in vivo studies, is crucial for maintaining the phenotype of cartilage. However, excessive or insufficient loading has deleterious effects and promotes the onset of cartilage degeneration. Informed by the prominent role of biomechanics, mechanical stimuli have been harnessed to enhance redifferentiation of chondrocytes and chondroinduction of other cell types, thus providing new chondrocyte cell sources. Biomechanical stimuli, such as hydrostatic pressure or compression, have been used to enhance the functional properties of neocartilage. By identifying pathways involved in mechanical stimulation, chemical equivalents that mimic mechanical signaling are beginning to offer exciting new methods for improving neocartilage. Harnessing biomechanics to improve differentiation, maintenance, and regeneration is emerging as pivotal toward producing functional neocartilage that could eventually be used to treat cartilage degeneration.


Office-based management of opioid dependence with buprenorphine: clinical practices and barriers.
Buprenorphine is a safe, effective and underutilized treatment for opioid dependence that requires special credentialing, known as a waiver, to prescribe in the United States. To describe buprenorphine clinical practices and barriers among office-based physicians. Cross-sectional survey. Two hundred thirty-five office-based physicians waivered to prescribe buprenorphine in Massachusetts. Questionnaires mailed to all waivered physicians in Massachusetts in October and November 2005 included questions on medical specialty, practice setting, clinical practices, and barriers to prescribing. Logistic regression analyses were used to identify factors associated with prescribing. Prescribers were 66% of respondents and prescribed to a median of ten patients. Clinical practices included mandatory counseling (79%), drug screening (82%), observed induction (57%), linkage to methadone maintenance (40%), and storing buprenorphine notes separate from other medical records (33%). Most non-prescribers (54%) reported they would prescribe if barriers were reduced. Being a primary care physician compared to a psychiatrist (AOR: 3.02; 95% CI: 1.48-6.18) and solo practice only compared to group practice (AOR: 3.01; 95% CI: 1.23-7.35) were associated with prescribing, while reporting low patient demand (AOR: 0.043, 95% CI: 0.009-0.21) and insufficient institutional support (AOR: 0.37; 95% CI: 0.15-0.89) were associated with not prescribing. Capacity for increased buprenorphine prescribing exists among physicians who have already obtained a waiver to prescribe. Increased efforts to link waivered physicians with opioid-dependent patients and initiatives to improve institutional support may mitigate barriers to buprenorphine treatment. Several guideline-driven practices have been widely adopted, such as adjunctive counseling and monitoring patients with drug screening.


Capture of kinesthesis by a competing cutaneous input.
In four experiments, blindfolded participants were presented with pairs of stimuli simultaneously, one to each index finger. Participants moved one index finger, which was presented with cutaneous and/or kinesthetic stimuli, and this movement caused a raised line to move underneath the other, stationary index finger in a yoked manner. The stimuli were 180º rotations of each other (e.g., < and >), and thus when a < was traced with the moving finger, it caused a > to be felt at the stationary finger. When asked to report the experience, participants predominantly reported the cutaneous stimulus, seemingly being ignorant of the kinesthetic stimulus. This appears to be an intrahaptic capture phenomenon, which is of interest because it suggests that conflict between intrahaptic sensory stimuli can go unnoticed; sometimes we are unaware of how we moved, and sometimes we do not know what we touched. The results are interpreted in light of optimal integration, perceptual suppression, reafference suppression, and inattentional blindness.


Long noncoding RNA TUG1 is downregulated in non-small cell lung cancer and can regulate CELF1 on binding to PRC2.
Long noncoding RNAs (lncRNAs) play crucial roles in tumorigenesis, and lncRNA taurine-upregulated gene 1 (TUG1) has been proven to be associated with several human cancers. However, the mechanisms of TUG1-involved regulation remain largely unknown. We examined the expressions of TUG1 in a cohort of 89 patients with non-small cell lung cancer (NSCLC) to determine the association between TUG1 expression and clinical parameters. We used circular chromosome conformation capture (4C) coupled with next-generation sequencing to explore the genome regions that interact with TUG1 and the TUG1-mediated regulation. TUG1 was significantly downregulated, and the TUG1 downregulation correlated with sex (p = 0.006), smoking status (p = 0.016), and tumor differentiation grade (p = 0.001). Knockdown of TUG1 significantly promoted the proliferation of NSCLC cells. According to the bioinformatic analysis result of TUG1 4C sequencing data, 83 candidate genes and their interaction regions were identified. Among these candidate genes, CUGBP and Elav-like family member 1 (CELF1) are potential targets of TUG1 in-trans regulation. To confirm the interaction between TUG1 and CELF1, relative expressions of CELF1 were examined in TUG1 knockdown H520 cells; results showed that CELF1 was significantly upregulated in TUG1 knockdown H520 cells. RNA immunoprecipitation was then performed to examine whether TUG1 RNA was bound to PRC2, a TUG1-involved regulation mechanism reported in previous studies. The results demonstrated that TUG1 RNA was bound to enhancer of zeste protein 2/embryonic ectoderm development (EZH2/EED), which is essential for PRC2. Finally, our designed ChIP assay revealed that the EZH2/EED was bound to the promotor region of CELF1 within 992 bp upstream of the transcript start site. TUG1 is downregulated in NSCLC. Using TUG1 4C sequencing and bioinformatic analysis, we found CELF1 to be a potential target of TUG1 RNA in in-trans regulation. Moreover, subsequent experiments showed that TUG1 RNA could bind to PRC2 in the promotor region of CELF1 and negatively regulate CELF1 expressions in H520 cells. Our results may facilitate developing new treatment modalities targeting TUG1/PRC2/CELF1 interactions in patients with NSCLC.


Analyzing Circle of Willis blood flow in ischemic stroke patients through 3D Stroke Arterial Flow Estimation.
Background The objective of ischemic stroke (IS) treatment is to achieve revascularization in cerebral arteries to restore blood flow. However, there is no available method to extract arterial flow data from clinical CTA images. We developed 3D Stroke Arterial Flow Estimation (SAFE), which provides blood flow data throughout the Circle of Willis based on 3D CTA and allows comparison of arterial flow distribution in the brain. Methods We implemented a newly developed 3D vascular reconstruction algorithm for clinical stroke CTA images. Based on the patient-specific vascular structure, SAFE calculates time-resolved blood flow information for the entire Circle of Willis and allows quantitative flow study of IS cases. Clinical IS cases are presented to demonstrate the feasibility. Four patients with CTA images and CT perfusion data were studied. To validate the SAFE analysis, correlation analysis comparing blood flow at the MCA, ICA, and BA was performed. Results Different blood flow patterns were found in individual IS patients. Altered flow patterns and high collateral flow rates were found near occlusions in all cases. Quantitative comparison of blood flow data showed that SAFE obtained flow data and CTP were significantly correlated and provide complementary information about cerebral blood flow for individual patients. Conclusions We present SAFE analysis for collecting detailed time-resolved cerebral arterial flow data in the entire Circle of Willis for IS. Further study with more cases may be important to test the clinical utilization of SAFE and helpful to the study of the underlying hemodynamics of stroke.


Facilitation across stress gradients: the importance of local adaptation.
While there is some information on genetic variation in response to competition in plants, we know nothing about intraspecific variation in facilitation. Previous studies suggest that facilitation should increase fitness in stressful environments. However, whether a plant experiences an environment as stressful may depend on prior adaptive responses to stressors at a site. Local adaptation to stress at a site may reduce the likelihood of facilitation. Seeds of Plantago erecta from stressful (serpentine soil) and non-stressful (non-serpentine soil) edaphic environments were reciprocally planted into these two soil types. Although competition did not differ significantly among seed sources, there was evidence for a local adaptation effect on facilitation. Non-serpentine seeds planted into serpentine soil exhibited greater individual plant biomass at higher densities. The interaction between population source and growth environment indicates a role for evolutionary processes such as local adaptation in the expression of facilitation in plants.


A Review of Multimodal Medical Image Fusion Techniques.
The medical image fusion is the process of coalescing multiple images from multiple imaging modalities to obtain a fused image with a large amount of information for increasing the clinical applicability of medical images. In this paper, we attempt to give an overview of multimodal medical image fusion methods, putting emphasis on the most recent advances in the domain based on (1) the current fusion methods, including based on deep learning, (2) imaging modalities of medical image fusion, and (3) performance analysis of medical image fusion on mainly data set. Finally, the conclusion of this paper is that the current multimodal medical image fusion research results are more significant and the development trend is on the rise but with many challenges in the research field.


Current Progress on Assessing the Prognosis for Anti-N-Methyl-D-Aspartate Receptor (NMDAR) Encephalitis.
Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is the most common antineuronal antibody encephalitis in autoimmune encephalitis found at present. It has complex clinical manifestations such as psychiatric and behavioral abnormalities, seizures, movement disorders, consciousness, and autonomic dysfunction. The relationship between those manifestations and prognosis is unclear. Electroencephalography (EEG) is gradually becoming useful in the evaluation of the prognosis of anti-NMDAR encephalitis patients, while imaging and antibody testing have a limited prognostic value. Starting the patients on adequate treatments (such as immunotherapy) in a timely fashion has a positive impact on their prognosis. Nevertheless, research on the prognosis of anti-NMDAR encephalitis remains scarce. Here, we review the current advances of prognosis-related research from the clinical manifestations of the disease and auxiliary examinations such as EEG, magnetic resonance imaging (MRI), 18F fluorodeoxyglucose positron emission tomography (FDG-PET), and antibody measurement. In addition, we also discuss the impact of different treatment options on prognosis. In-depth research on the prognosis of patients with anti-NMDAR encephalitis will contribute to a better understanding of this disease, leading to better treatments options and, ultimately, a better prognosis.


Why is premorbid BMI consistently elevated in clinical samples, but not in risk factor samples, of individuals with eating disorders?
Body image disturbance is widely viewed as contributing to the development and maintenance of disordered eating. Yet this perspective is not inconsistent with the possibility that elevated premorbid BMIs also increase the risk of developing eating disorders. Research examining whether actual body size may play a role in eating disorder development reveals a curious pattern of findings. Few prospective risk factor studies conducted with community-based samples found a relationship between premorbid BMI and subsequent eating disorder pathology whereas retrospective research conducted with clinical samples indicates a consistent pattern of elevated premorbid BMIs relative to population norms or control groups. This study documents these disparate findings, considers potential explanations for them and proposes further study of premorbid BMI as a factor contributing to the psychopathology of eating disorders, particularly among those who come to the attention of treatment providers.


Cell cycle analysis of retroviral vector gene expression during early infection.
To examine the pattern of retroviral vector gene expression during early stages of infection, a Moloney murine leukemia virus (M-MuLV)-based vector that transcribes the simian virus 40 (SV40) large T antigen (Tag) gene from the viral long terminal repeat (LTR) was used to infect proliferating rodent fibroblasts. At various times after infection, cells were fixed and stained for Tag by indirect immunofluorescence, and for DNA using propidium iodide. Tag immunofluorescence and DNA content were both quantified by flow cytometry. The results showed that Tag expression was first detected exclusively in the late G1 and early S phases of the cell cycle, approximately 12 h postinfection. The infection was synchronous in that Tag-expressing cells detected at 12 h, in late G1 and early S, moved as a discrete population through S phase, into the G2 + M phases of the cell cycle and then back into G1 during the next 8-10 h. The presence of a synchronous Tag-expressing cell population suggests that, at time of infection, cells in certain phases of the cell cycle were more susceptible to infection than cells in other phases. This may be related to synchronizing events that must occur before viral genes are expressed in infected cells; one such event may be integration of viral DNA into cellular chromosomes (i.e., provirus formation) that requires cells to pass through an M phase.


Vascular endothelial growth factor-d modulates caliber and function of initial lymphatics in the dermis.
The lymphatic vasculature is important for skin biology as it maintains dermal fluid homeostasis. However, the molecular determinants of the form and function of the lymphatic vasculature in skin are poorly understood. Here, we explore the role of vascular endothelial growth factor-d (Vegf-d), a lymphangiogenic glycoprotein, in determining the form and function of the dermal lymphatic network, using Vegf-d-deficient mice. Initial lymphatic vessels in adult Vegf-d-deficient mice were significantly smaller than wild-type but collecting lymphatics were unaltered. The uptake/transport of dextran in initial lymphatics of Vegf-d-deficient mice was far less efficient, indicating compromised function of these vessels. The role of Vegf-d in modulating initial lymphatics was further supported by delivery of Vegf-d in skin of wild-type mice, which promoted enlargement of these vessels. Vegf-d-deficient mice were subjected to cutaneous wounding to challenge lymphatic function: the resulting wound epithelium was highly edematous and thicker, reflecting inadequate lymphatic drainage. Unexpectedly, myofibroblasts were more abundant in Vegf-d-deficient wounds leading to faster wound closure, but resorption of granulation tissue was compromised suggesting poorer-quality healing. Our findings demonstrate that Vegf-d deficiency alters the caliber of initial lymphatics in the dermis leading to reduced functional capacity.


Paracrine factors from mesenchymal stem cells attenuate epithelial injury and lung fibrosis.
Paracrine factors are currently considered to be the major mechanism through which mesenchymal stem cells (MSCs) exert their actions. The aim of this study was to investigate the protective effects of conditioned medium (CM) from bone marrow mesenchymal stem cells (MSC) on bleomycin (BLM)‑induced lung injury and fibrosis, both in vitro and in vivo. A549 human non‑small cell lung cancer epithelial cells were cultured in serum‑free medium, or MSC‑CM, both with or without BLM. The protective effects of MSC‑CM was determined by MTT assay to assess cell viability and Annexin V‑PE to assess apoptosis. Rats were intratracheally injected with MSC‑CM, saline, or conditioned medium from fibroblasts on day 0 and day 3 after intratracheal administration of BLM, and were sacrificed on day 28. Lung injury and fibrosis were assessed by histological assessment, Ashcroft score, and hydroxyproline assay; lung cell apoptosis was detected using terminal deoxynucleotidyl transferase dUTP nick end labeling assay. In comparison to the control group (0.17±0.01), 8 and 16% MSC‑CM had a significant stimulatory effect on A549 cellular proliferation (0.24±0.03 and 0.24±0.04, respectively, P<0.01). A549 cells cultured with MSC‑CM were protected from BLM‑induced apoptosis, 23.43±3.76% vs. 38.06±4.32%; (P<0.05). In the BLM‑challenged rats, MSC‑CM was shown to protect against lung fibrosis in terms of lung inflammation, fibrotic scores, collagen deposition, and cell apoptosis. This data suggests that MSCs are capable of protecting against lung injury and fibrosis both in vitro and in vivo through a paracrine anti‑inflammatory mechanism. MSC‑CM may provide a novel approach for the treatment of lung fibrosis.


The pharmacology of selective inhibition of COX-2.
Selective inhibitors of cyclooxygenase (COX)-2 were developed to improve the safety of anti-inflammatory therapy in patients at elevated risk for gastrointestinal complications which are thought to be caused primarily by depression of COX-1 derived mucosal prostanoids. They were not expected to be more efficacious analgesics than compounds acting on both cyclooxygenases, the traditional (t) non-steroidal antiinflammatory drugs (NSAIDs). While these predictions were generally supported by clinical evidence, an elevated rate of severe cardiovascular complications was observed in randomized controlled trials of three chemically distinct COX-2 selective compounds. The cardiovascular hazard is plausibly explained by the depression of COX-2 dependent prostanoids formed in vasculature and kidney; vascular prostacyclin (PGI2) constrains the effect of prothrombotic and atherogenic stimuli, and renal medullary prostacyclin and prostaglandin (PG) E(2) formed by COX-2 contribute to arterial pressure homeostasis. A drug development strategy more closely linking research into the biology of the drug target with clinical drug development may have allowed earlier recognition of these mechanisms and the cardiovascular risk of COX-2 inhibition. Open questions are i) whether the gastrointestinal benefit of COX-2 selective compounds drugs can be conserved by identifying individuals at risk and excluding them from treatment; ii) whether the risk extends to tNSAIDs; iii) and whether alternative strategies to anti-inflammatory therapy with a more advantageous risk-benefit profile can be developed.


Water-lactose behavior as a function of concentration and presence of lactic acid in lactose model systems.
The presence of high amounts of lactic acid in acid whey restricts its ability to be further processed because lactose appears to remain in its amorphous form. A systematic study is lacking in this regard especially during the concentration step. Hence, the main aim of the study was to establish the structure and behavior of water molecules surrounding lactose in the presence of 1% (wt/wt) lactic acid at a concentration up to 50% (wt/wt). Furthermore, the crystallization nature of freeze-dried lactose with or without lactic acid was established using differential scanning calorimetry and Fourier transform infrared spectroscopy. Two mechanisms were proposed to describe the behavior of water molecules around lactose molecules during the concentration of pure lactose and lactose solutions with lactic acid. Pure lactose solution exhibited a water evaporation enthalpy of ~679 J·g(-1), whereas lactose+ lactic acid solution resulted in ~965 J·g(-1) at a 50% (wt/wt) concentration. This indicates a greater energy requirement for water removal around lactose in the presence of lactic acid. Higher crystallization temperatures were observed with the presence of lactic acid, indicating a delay in crystallization. Furthermore, less crystalline lactose (~12%) was obtained in the presence of lactic acid, indicating high amorphous nature compared with pure lactose where ~50% crystallinity was obtained. The Fourier transform infrared spectra revealed that the strong hydration layer consisting lactic acid and H3O(+) ions surrounded lactose molecules via strong H bonds, which restricted water mobility, induced a change in structure of lactose, or both, creating unfavorable conditions for lactose crystallization. Thus, partial or complete removal of lactic acid from acid whey may be the first step toward improving the ability of acid whey to be processed.