The role of DMPK science in improving pharmaceutical research and development efficiency.
The successful regulatory authority approval rate of drug candidates in the drug development pipeline is crucial for determining pharmaceutical research and development (R&D) efficiency. Regulatory authorities include the US Food and Drug Administration (FDA), European Medicines Agency (EMA), and Pharmaceutical and Food Safety Bureau Japan (PFSB), among others. Optimal drug metabolism and pharmacokinetics (DMPK) properties influence the progression of a drug candidate from the preclinical to the clinical phase. In this review, we provide a comprehensive assessment of essential concepts, methods, improvements, and challenges in DMPK science and its significance in drug development. This information provides insights into the association of DMPK science with pharmaceutical R&D efficiency.


Laue lens for radiotherapy applications through a focused hard x-ray beam: a feasibility study on requirements and tolerances.
Focusing a hard x-ray beam would represent an innovative technique for tumour treatment, since such a beam may deliver a dose to a tumour located at a given depth under the skin, sparing the surrounding healthy cells. A detailed study of a focusing system for hard x-ray aimed at radiotherapy is presented here. Such a focusing system, named Laue lens, exploits x-ray diffraction and consists of a series of crystals disposed as concentric rings capable of concentrating a flux of x-rays towards a focusing point. A feasibility study regarding the positioning tolerances of the crystalline optical elements has been carried out. It is shown that a Laue lens can effectively be used in the context of radiotherapy for tumour treatments provided that the mounting errors are below certain values, which are reachable in the modern micromechanics. An extended survey based on an analytical approach and on simulations is presented for precisely estimating all the contributions of each mounting error, analysing their effect on the focal spot of the Laue lens. Finally, a simulation for evaluating the released dose in a water phantom is shown.


Pedestrian activity among California adults.
The objectives of this study were to assess (1) pedestrian activity levels among adults, (2) where and why adults engage in pedestrian activity, and (3) what adults consider when deciding where to engage in pedestrian activity. Pedestrian activity was assessed in 12,036 California adults, >or=18 years, using a random digit-dial telephone survey. Significant differences were identified by race, sex, age, and physical activity level in the type, location, and purpose of pedestrian activities. Men engage in pedestrian activity at work, and women engage in pedestrian activity while escorting children to school and running errands. Whites primarily engage in leisure-time pedestrian activity, and nonwhites are more likely to engage in pedestrian activity for transportation. Older adults were less active than their younger counterparts. These findings should be considered by public health agencies and their partners as they continue to increase and promote opportunities for pedestrian activity. Additional research is needed to assess older adults' physical activity patterns and preferences, barriers, and facilitators to effectively tailor physical activity promotion efforts to this at-risk group.


Ribosome-targeting antibiotic control NLRP3-mediated inflammation by inhibiting mitochondrial DNA synthesis.
While antibiotics are designed to target bacteria specifically, most are known to affect host cell physiology. Certain classes of antibiotics have been reported to have immunosuppressive effects, but the underlying mechanisms remain elusive. Here, we show that doxycycline, a ribosomal-targeting antibiotic, effectively inhibited both mitochondrial translation and nucleotide-binding domain and leucine-rich repeat-containing protein 3 (NLRP3) inflammasome-mediated caspase-1 activation and interleukin-1β (IL-1β) production in bone-marrow-derived macrophages (BMDMs). In addition, knockdown of mitochondrial methionyl-tRNA formyltransferase (Mtfmt), which is rate limiting for mitochondrial translation, also resulted in the inhibition of NLRP3 inflammasome-mediated caspase-1 activation and IL-1β secretion. Furthermore, both doxycycline treatment and Mtfmt knockdown blocked the synthesis of mitochondrial DNA (mtDNA) and the generation of oxidized mtDNA (Ox-mtDNA), which serves as a ligand for NLRP3 inflammasome activation. In addition, in vivo results indicated that doxycycline mitigated NLRP3 inflammasome-dependent inflammation, including lipopolysaccharide-induced systemic inflammation and endometritis. Taken together, the results unveil the antibiotics targeting the mitoribosome have the ability to mitigate NLRP3 inflammasome activation by inhibiting mitochondrial translation and mtDNA synthesis thus opening up new possibilities for the treatment of NLRP3-related diseases.


Type 2 diabetes: a case study.
Increased prevalence of diabetes in the community has been accompanied by an increase in diabetes in hospitalised patients. About a quarter of these patients experience a hypoglycaemic episode during their admission, which is associated with increased risk of mortality and length of stay. This article examines the aetiology, pathophysiology, diagnosis and treatment of type 2 diabetes using a case study approach. The psychosocial implications for the patient are also discussed. The case study is based on a patient with diabetes who was admitted to hospital following a hypoglycaemic episode and cared for during a practice placement. The importance of early diagnosis of diabetes and the adverse effects of delayed diagnosis are discussed.


Posttranslational modifications of ACE2 protein: Implications for SARS-CoV-2 infection and beyond.
The present worldwide pandemic of coronavirus disease 2019 (COVID-19) has highlighted the important function of angiotensin-converting enzyme 2 (ACE2) as a receptor for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) entry. A deeper understanding of ACE2 could offer insights into the mechanisms of SARS-CoV-2 infection. While ACE2 is subject to regulation by various factors in vivo, current research in this area is insufficient to fully elucidate the corresponding pathways of control. Posttranslational modification (PTM) is a powerful tool for broadening the variety of proteins. The PTM study of ACE2 will help us to make up for the deficiency in the regulation of protein synthesis and translation. However, research on PTM-related aspects of ACE2 remains limited, mostly focused on glycosylation. Accordingly, a comprehensive review of ACE2 PTMs could help us better understand the infection process and provide a basis for the treatment of COVID-19 and beyond.


Explaining the Microtubule Energy Balance: Contributions Due to Dipole Moments, Charges, van der Waals and Solvation Energy.
Microtubules are the main components of mitotic spindles, and are the pillars of the cellular cytoskeleton. They perform most of their cellular functions by virtue of their unique dynamic instability processes which alternate between polymerization and depolymerization phases. This in turn is driven by a precise balance between attraction and repulsion forces between the constituents of microtubules (MTs)-tubulin dimers. Therefore, it is critically important to know what contributions result in a balance of the interaction energy among tubulin dimers that make up microtubules and what interactions may tip this balance toward or away from a stable polymerized state of tubulin. In this paper, we calculate the dipole-dipole interaction energy between tubulin dimers in a microtubule as part of the various contributions to the energy balance. We also compare the remaining contributions to the interaction energies between tubulin dimers and establish a balance between stabilizing and destabilizing components, including the van der Waals, electrostatic, and solvent-accessible surface area energies. The energy balance shows that the GTP-capped tip of the seam at the plus end of microtubules is stabilized only by - 9 kcal/mol, which can be completely reversed by the hydrolysis of a single GTP molecule, which releases + 14 kcal/mol and destabilizes the seam by an excess of + 5 kcal/mol. This triggers the breakdown of microtubules and initiates a disassembly phase which is aptly called a catastrophe.


The Electronic Structure of Amorphous Carbon Nanodots.
We have studied hydrogen-passivated amorphous carbon nanostructures with semiempirical molecular orbital theory in order to provide an understanding of the factors that affect their electronic properties. Amorphous structures were first constructed using periodic calculations in a melt/quench protocol. Pure periodic amorphous carbon structures and their counterparts doped with nitrogen and/or oxygen feature large electronic band gaps. Surprisingly, descriptors such as the elemental composition and the number of sp(3)-atoms only influence the electronic structure weakly. Instead, the exact topology of the sp(2)-network in terms of effective conjugation defines the band gap. Amorphous carbon nanodots of different structures and sizes were cut out of the periodic structures. Our calculations predict the occurrence of localized electronic surface states, which give rise to interesting effects such as amphoteric reactivity and predicted optical band gaps in the near-UV/visible range. Optical and electronic gaps display a dependence on particle size similar to that of inorganic colloidal quantum dots.


[Isolation of the Tahyña bunyavirus in the Arctic].
Tahyna virus (Bunyaviridae, Bunyavirus, the California encephalitis complex) was isolated from Aedes communis complex mosquitoes collected at the border of the north-taiga landscape zone (in latitude 68 degrees North and longitude 33 degrees East) at the Kolsky peninsula (the Murmansk region). The LEIV-9843 Mur strain was isolated from 2.4 thousand mosquitoes collected there (altogether 3.8 thousand mosquitoes had been collected in the Murmansk region). This is the first isolation in the USSR of a California complex virus in the Arctic and the northernmost site of Tahyna virus isolation in the world. 18% of the human population residing near the site of the virus isolation had virus-neutralizing antibody to Tahyna virus.


A biodistribution study of PEGylated PCL-based nanoparticles in C57BL/6 mice bearing B16/F10 melanoma.
One of the major drawbacks that limits the clinical application of nanoparticles is the lack of preliminary investigations related to their biocompatibility, biodegradability and biodistribution. In this work, biodegradable PEGylated polymer nanoparticles (NPs) have been synthesized by using macromonomers based on poly(ε-caprolaconte) oligomers. More in detail, NPs have been produced by adopting a surfactant-free semibatch emulsion polymerization process using PEG chains as a stabilizing agent. The NPs were also labeled with rhodamine B covalently bound to the NPs to quantitatively study their biodistribution in vivo. NPs were investigated in both in vitro and in vivo preclinical systems to study their biodistribution in mice bearing B16/F10 melanoma, as well as their biocompatibility and biodegradability. The NP concentration was evaluated in different tissues at several times after intravenous injection. The disappearance of the NPs from the plasma was biphasic, with distribution and elimination half-lives of 30 min and 15 h, respectively. NPs were retained in tumors and in filter organs for a long time, were still detectable after 7 d and maintained a steady concentration in the tumor for 120 h. 48 h after injection, 70 ± 15% of the inoculated NPs were excreted in the feces. The favorable tumor uptake, fast excretion and absence of cytotoxicity foster the further development of produced NPs as drug delivery carriers.


Health in China. From Mao to market reform.
After the Liberation by Mao Ze Dong's Communist army in 1949, China experienced massive social and economic change. The dramatic reductions in mortality and morbidity of the next two decades were brought about through improvements in socioeconomic conditions, an emphasis on prevention, and almost universal access to basic health care. The economic mismanagement of the Great Leap Forward brought about a temporary reversal in these positive trends. During the Cultural Revolution there was a sustained attack on the privileged position of the medical profession. Most city doctors were sent to work in the countryside, where they trained over a million barefoot doctors. Deng Xiao Ping's radical economic reforms of the late 1970s replaced the socialist system with a market economy. Although average incomes have increased, the gap between rich and poor has widened.


ETV5 regulates sertoli cell chemokines involved in mouse stem/progenitor spermatogonia maintenance.
Spermatogonial stem cells are the only stem cells in the body that transmit genetic information to offspring. Although growth factors responsible for self-renewal of these cells are known, the factors and mechanisms that attract and physically maintain these cells within their microenvironment are poorly understood. Mice with targeted disruption of Ets variant gene 5 (Etv5) show total loss of stem/progenitor spermatogonia following the first wave of spermatogenesis, resulting in a Sertoli cell-only phenotype and aspermia. Microarray analysis of primary Sertoli cells from Etv5 knockout (Etv5(-/-)) versus wild-type (WT) mice revealed significant decreases in expression of several chemokines. Chemotaxis assays demonstrated that migration of stem/progenitor spermatogonia toward Etv5(-/-) Sertoli cells was significantly decreased compared to migration toward WT Sertoli cells. Interestingly, differentiating spermatogonia, spermatocytes, and round spermatids were not chemoattracted by WT Sertoli cells, whereas stem/progenitor spermatogonia showed a high and significant chemotactic index. Rescue assays using recombinant chemokines indicated that C-C-motif ligand 9 (CCL9) facilitates Sertoli cell chemoattraction of stem/progenitor spermatogonia, which express C-C-receptor type 1 (CCR1). In addition, there is protein-DNA interaction between ETV5 and Ccl9, suggesting that ETV5 might be a direct regulator of Ccl9 expression. Taken together, our data show for the first time that Sertoli cells are chemoattractive for stem/progenitor spermatogonia, and that production of specific chemokines is regulated by ETV5. Therefore, changes in chemokine production and consequent decreases in chemoattraction by Etv5(-/-) Sertoli cells helps to explain stem/progenitor spermatogonia loss in Etv5(-/-) mice.


[Suprasegmental speech in prelingual hearing-impaired children with cochlear implants, hearing-impaired children without implants and normal hearing children].
A review of the relevant literature indicates a lack of knowledge regarding suprasegmental speech characteristics in prelingual hearing impaired children with cochlear implants. This study is aimed at examining certain suprasegmental features in the speech of prelingual hearing impaired children with cochlear implants by perceptual ratings and acoustic analyses, comparing these results to that of prelingual hearing impaired children without implants, and normal hearing children of the same age. Twelve Afrikaansspeaking children between the ages of six and ten years were included in three groups. An integrated view of the perceptual and acoustic results shows that the cochlear implanted children's suprasegmental speech characteristics bear closer resemblance to those of the normal hearing group than those of the hearing impaired group.


Macrophage 12(S)-HETE Enhances Angiotensin II-Induced Contraction by a BLT2 (Leukotriene B4 Type-2 Receptor) and TP (Thromboxane Receptor)-Mediated Mechanism in Murine Arteries.
12/15-LO (12/15-lipoxygenase), encoded by Alox15 gene, metabolizes arachidonic acid to 12(S)-HETE (12-hydroxyeicosatetraenoic acid). Macrophages are the major source of 12/15-LO among immune cells, and 12/15-LO plays a crucial role in development of hypertension. Global Alox15- or macrophage-deficient mice are resistant to Ang II (angiotensin II)-induced hypertension. This study tests the hypothesis that macrophage 12(S)-HETE contributes to Ang II-mediated arterial constriction and thus to development of Ang II-induced hypertension. Ang II constricted isolated abdominal aortic and mesenteric arterial rings. 12(S)-HETE (100 nmol/L) alone was without effect; however, it significantly enhanced Ang II-induced constriction. The presence of wild-type macrophages also enhanced the Ang II-induced constriction, while Alox15-/- macrophages did not. Using this model, pretreatment of aortic rings with inhibitors, receptor agonists/antagonists, or removal of the endothelium, systematically uncovered an endothelium-mediated, Ang II receptor-2-mediated and superoxide-mediated enhancing effect of 12(S)-HETE on Ang II constrictions. The role of superoxide was confirmed using aortas from p47phox-/- mice where 12(S)-HETE failed to enhance constriction to Ang II. In cultured arterial endothelial cells, 12(S)-HETE increased the production of superoxide, and 12(S)-HETE or Ang II increased the production of an isothromboxane-like metabolite. A TP (thromboxane receptor) antagonist inhibited 12(S)-HETE enhancement of Ang II constriction. Both Ang II-induced hypertension and the enhancing effect of 12(S)-HETE on Ang II contractions were eliminated by a BLT2 (leukotriene B4 receptor-2) antagonist. These results outline a mechanism where the macrophage 12/15-LO pathway enhances the action of Ang II. 12(S)-HETE, acting on the BLT2, contributes to the hypertensive action of Ang II in part by promoting endothelial synthesis of a superoxide-derived TP agonist.


Single-Step Synthesis of NiI from NiII with H2.
Chemists have long sought to regulate the reactivity of H2 , to yield hydride ions, hydrogen atoms, or electrons on demand. One source of inspiration for achieving this control is [NiFe]hydrogenase ([NiFe]H2 ase), which reacts with H2 to form various hydrogen active species such as NiIII hydride species, NiII hydride species, and NiI low-valent species. Chemists have attempted to synthesize these hydrogen active species not only as models for the active species of [NiFe]H2 ase, but also as electron transfer catalysts. However, the synthesis of NiI complex directly from H2 has not been reported. This paper reports the first example of a single-step synthesis of a NiI complex, via reaction of a NiII complex with H2 , stable for over 3 months at room temperature and we further demonstrate a reductive coupling of acridinium ions as part of a reaction cycle.


Carbon catabolite repression of the maltose transporter revealed by X-ray crystallography.
Efficient carbon utilization is critical to the survival of microorganisms in competitive environments. To optimize energy usage, bacteria have developed an integrated control system to preferentially uptake carbohydrates that support rapid growth. The availability of a preferred carbon source, such as glucose, represses the synthesis and activities of proteins necessary for the transport and metabolism of secondary carbon sources. This regulatory phenomenon is defined as carbon catabolite repression. In enteric bacteria, the key player of carbon catabolite repression is a component of the glucose-specific phosphotransferase system, enzyme IIA (EIIA(Glc)). It is known that unphosphorylated EIIA(Glc) binds to and inhibits a variety of transporters when glucose is available. However, understanding the underlying molecular mechanism has been hindered by the complete absence of structures for any EIIA(Glc)-transporter complexes. Here we present the 3.9 Å crystal structure of Escherichia coli EIIA(Glc) in complex with the maltose transporter, an ATP-binding cassette (ABC) transporter. The structure shows that two EIIA(Glc) molecules bind to the cytoplasmic ATPase subunits, stabilizing the transporter in an inward-facing conformation and preventing the structural rearrangements necessary for ATP hydrolysis. We also show that the half-maximal inhibitory concentrations of the full-length EIIA(Glc) and an amino-terminal truncation mutant differ by 60-fold, consistent with the hypothesis that the amino-terminal region, disordered in the crystal structure, functions as a membrane anchor to increase the effective EIIA(Glc) concentration at the membrane. Together these data suggest a model of how the central regulatory protein EIIA(Glc) allosterically inhibits maltose uptake in E. coli.


Foot pigmentation in Blacks.
Pigmentation of the soles of the feet was studied during postmortem examination of 70 Black patients. Forty-eight (57%) showed some pigmentation (grade II) of the soles of the feet and 4 (6%) showed definite black spots, i.e. focal areas of dense black pigmentation (grade III). A distinction between clinical grade II or grade III lesions could not be made on the basis of microscopic appearance. No true naeyi were seen. Increases of melanin pigmentation and minor degrees of melanocytic proliferation were all that was apparent. Although the development of malignant melanoma may be related to the presence of black spots on the feet of darkly pigmented races, all such spots are certainly not naevi.


The cystic fibrosis lung microbiome.
The chronic colonization of the lower airways by bacterial pathogens is the leading cause of morbidity and mortality in patients with cystic fibrosis (CF). Pseudomonas aeruginosa is the most common CF pathogen, followed by Staphylococcus aureus. Improvements in airway clearance and more effective treatment of the conventional CF pathogens has led to the emergence of new airway pathogens such as Stenotrophomonas maltophilia, Mycobacterium abscessus, and Achromobacter. More recently, it has become appreciated that the lower airways in patients with CF are colonized by a more complex polymicrobial community composed primarily of bacteria found in the upper respiratory tract. This includes obligate anaerobes, most commonly Prevotella. Expanded culturing methods and culture-independent molecular methods are being used to characterize the composition and dynamics of these polymicrobial communities in patients with CF. The contribution of the CF microbiome to airway disease is actively being investigated and will present new opportunities for disease management in CF. However, there remain many challenges that must be overcome if microbiome profiling is going to inform clinical practice.


Meaning in life following service among post-9/11 military veterans: A latent growth mixture model analysis.
Meaning in life refers to the "sense made of, and significance felt regarding, the nature of one's being and existence." Meaningful living promotes well-being, resilience, and personal growth. Yet, much remains unknown about how meaning changes over time and determinants of meaning, particularly during major life transitions. We identified distinct trajectories of meaning using latent growth mixture models and examined prospective predictors of class membership in a military veteran cohort assessed at multiple time points throughout the first 3 years after leaving service. Three trajectories were identified: consistently high meaning (89.5%; n = 7025), diminishing meaning (6.1%; n = 479), and strengthening meaning (4.4%; n = 348). Veterans with greater posttraumatic stress symptoms, depression symptoms, and moral injury experienced increased odds of a less adaptive trajectory (i.e. diminishing and/or strengthening vs. consistently high meaning), whereas veterans who reported greater psychological resilience, community relationship satisfaction, and intimate relationship satisfaction experienced lower odds of a less adaptive trajectory. Several gender differences were also observed. Results provide insight into veteran subgroups that are more likely to experience lower meaning after leaving military service and thus may benefit from additional support to reduce their risk for poor longer-term health and well-being outcomes.


Early tobacco smoke exposure, preschool cool/hot inhibitory control, and young adolescents' externalizing/internalizing problems.
There is a dearth of research spanning multiple developmental stages to delineate the far-reaching implications of early tobacco smoke exposure for later behavior adaptation and also elucidate the underlying psychological mechanisms. Using NICHD SECCYD data, we conducted process model analyses to address this gap. Results indicated that early tobacco smoke exposure was not only negatively associated with preschool cool and hot inhibitory control, but also positively related to externalizing and internalizing problems in early adolescence. Further, early exposure was also positively associated with externalizing problems in early adolescence via a negative link with preschool cool inhibitory control. Behavior problems in toddlerhood also served as one mechanism underlying the implications of early exposure for behavior problems in early adolescence. As compared to nonexposed children, children with both prenatal and postnatal exposure tended to have lower cool and hot inhibitory control, whereas children with only prenatal exposure displayed lower hot inhibitory control. Regardless of whether exposure was across prenatal and postnatal periods, or restricted to prenatal phase alone, exposed children displayed more behavior problems in toddlerhood, which in turn predicted more behavior problems in early adolescence. All associations emerged after considering extensive potential confounding factors and did not vary across child sex. Taken together, early tobacco smoke exposure may induce self-regulation deficits and also early onset of behavior problems and thus elevate the risk of later psychopathology. Building a smoke free environment in pregnancy and infancy likely yields long-term benefits by facilitating adaptation in early adolescence. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Analysis of microspike movements on the neuronal growth cone.
Growth cones of chick sensory ganglion neurons in tissue culture were photographed at 60-sec intervals as they advanced over the substratum. Numbers of microspikes (or "filopodia") were recorded together with the time and position of their appearance, their rate of elongation, their lateral movements, their lifespan, and the position and manner of their disappearance. All microspikes go through cycles of extension, lateral movement, and shortening. These are irregular and unpredictable but show systematic differences depending on where on the growth cone they occur. At the leading edge of the growth cone microspike extension occurs at highest frequency and microspike shortening occurs at the lowest frequency; when the latter occurs in this region it often involves the advance of the margin of the cell in the form of a lamellipodium. Microspike loss occurs most often at the base of the growth cone, usually by the retraction of the microspike into the cell. Calculations of the gain and loss of microspikes at different regions of the growth cone show that they undergo a net retrograde flow, the rate of which is correlated with the forward advance of the growth cone. Individual microspikes can also move backward from the growth cone onto the axon (or "neurite"), an event that occurs most often on adhesive substrata. Our observations support a direct role of microspike movement in the advance of the growth cone. The primary force for axonal elongation appears to be the contraction of microspikes pulling the leading margin of the growth cone forward. At more proximal and peripheral regions of the growth cone, microspikes undergo a retrograde sweeping motion, followed by retraction into the cell, which may also contribute to the forward movement of the growth cone. We interpret these movements as arising from a flow of actin filaments and associated proteins which are incorporated into microspikes and lamellipodia at the leading edge of the growth cone, passing backward, and being deposited into the actin-rich membrane-associated cortex of the axonal cylinder.


Age-Based Comparison of Hematological Toxicity in Patients with Lung Cancer.
Because of the increasing age of the general population, there is an increasing number of older patients with lung cancer. Cancer chemotherapy often causes severe hematological toxicity in older patients. This study aimed to explore the risk factors affecting the hematological toxicity of cytotoxic anticancer drugs in patients with lung cancer. Data were retrospectively collected from 194 patients with lung cancer at Niigata University Medical and Dental Hospital, Japan, between April 2011 and March 2016, when the patients underwent their first round of cytotoxic chemotherapy. The patients were divided into three groups on the basis of age: <65, 65-74, and ≥75 years. Physiological functions and laboratory data before treatment, as well as hematological adverse events following chemotherapy, were compared among the groups. Patients aged ≥75 years were significantly more likely to experience grade 3 or 4 neutropenia, compared with patients aged <65 years. However, there were no differences in the incidence of anemia or thrombocytopenia among the age groups. The frequency of febrile neutropenia tended to increase with age. Multivariate analysis showed that age ≥75 years, male sex, and a performance status of ≥2 were independent factors for grade 3 or 4 neutropenia. Patients with 2 or 3 of these factors had a significantly higher frequency of neutropenia, compared with patients who had 0 or 1 of these factors. We found that age ≥75 years, male sex, and a performance status of ≥2 were independent risk factors for grade 3 or 4 neutropenia.


Genome-wide association study identifies Loci for body composition and structural soundness traits in pigs.
The recent completion of the swine genome sequencing project and development of a high density porcine SNP array has made genome-wide association (GWA) studies feasible in pigs. Using Illumina's PorcineSNP60 BeadChip, we performed a pilot GWA study in 820 commercial female pigs phenotyped for backfat, loin muscle area, body conformation in addition to feet and leg (FL) structural soundness traits. A total of 51,385 SNPs were jointly fitted using Bayesian techniques as random effects in a mixture model that assumed a known large proportion (99.5%) of SNPs had zero effect. SNP annotations were implemented through the Sus scrofa Build 9 available from pig Ensembl. We discovered a number of candidate chromosomal regions, and some of them corresponded to QTL regions previously reported. We not only have identified some well-known candidate genes for the traits of interest, such as MC4R (for backfat) and IGF2 (for loin muscle area), but also obtained novel promising genes, including CHCHD3 (for backfat), BMP2 (for loin muscle area, body size and several FL structure traits), and some HOXA family genes (for overall leg action). The candidate regions responsible for body conformation and FL structure soundness did not overlap greatly which implied that these traits were controlled by different genes. Functional clustering analyses classified the genes into categories related to bone and cartilage development, muscle growth and development or the insulin pathway suggesting the traits are regulated by common pathways or gene networks that exert roles at different spatial and temporal stages. This study is one of the earliest GWA reports on important quantitative traits in pigs, and the findings will contribute to the further biological function analysis of the identified candidate genes and potential utilization of them in marker assisted selection.


Management of allergic diseases in pregnancy.
Allergic diseases like asthma, allergic rhinitis, food allergy, hymenoptera allergy, or atopic dermatitis are highly prevalent in women of childbearing age and may affect up to 30% of this age group. This review focuses on the management of allergic diseases during pregnancy. Furthermore, we discuss the challenges of counseling women with allergic diseases in the reproductive age, including considerations relevant to the ongoing SARS-CoV-2 pandemic. To create the optimal milieu for the unborn child, a multitude of immunological changes occur during pregnancy which may favor type 2 responses and aggravate disease phenotypes. In co-occurrence with suboptimal preconception disease control, this elevated Th2 responses may aggravate allergic disease manifestations during pregnancy and pose a risk for mother and child. Due to limitations in conducting clinical trials in pregnant women, safety data on anti-allergic drugs during pregnancy are limited. The lack of information and concerns among pregnant patients demands counseling on the benefits of anti-allergic drugs and the potential and known risks. This includes information on the risk for mother and child of disease aggravation in the absence of treatment. By doing so, informed decisions and shared decision-making can take place.


Insomnia with objective short sleep duration is associated with deficits in neuropsychological performance: a general population study.
To examine the joint effect of insomnia and objective short sleep duration on neuropsychological performance. Representative cross-sectional study. Sleep laboratory. 1,741 men and women randomly selected from central Pennsylvania. None. Insomnia (n = 116) was defined by a complaint of insomnia with a duration > or = 1 year and the absence of sleep disordered breathing (SDB), while normal sleep (n = 562) was defined as the absence of insomnia, excessive daytime sleepiness, and SDB. Both groups were split according to polysomnographic sleep duration into 2 categories: > or = 6 h of sleep ("normal sleep duration") and < 6 h of sleep ("short sleep duration"). We compared the groups' performance on a comprehensive neuropsychological battery that measured processing speed, attention, visual memory, and verbal fluency, while controlling for age, race, gender, education, body mass index, and physical and mental health. No significant differences were detected between insomniacs and controls. However, the insomnia with short sleep duration group compared to the control with normal or short sleep duration groups showed poorer neuropsychological performance in variables such as processing speed, set-switching attention, and number of visual memory errors and omissions. In contrast, the insomnia with normal sleep duration group showed no significant deficits. Insomnia with objective short sleep duration is associated with deficits in set-switching attentional abilities, a key component of the "executive control of attention." These findings suggest that objective sleep duration may predict the severity of chronic insomnia, including its effect on neurocognitive function.


The pyramidal tract in congenital hemiparesis: relationship between morphology and function in periventricular lesions.
Three-dimensional MRI data sets were obtained from 12 young adult patients with congenital spastic hemiparesis caused by unilateral periventricular white matter lesions. The impact of these lesions on corticospinal projections to the upper and lower extremities was assessed on reconstructed semi-coronal planes following anatomical landmarks of somatotopic organization in the precentral gyrus and in the internal capsule: a more anterior plane running through the hand-knob of the precentral gyrus and the anterior portion of the posterior limb of the internal capsule representing projections to the upper extremity, and a more posterior plane running through the top of the precentral gyrus and the middle portion of the posterior limb of the internal capsule representing projections to the lower extremity. In addition, the total lesion extent was determined volumetrically, and Wallerian degeneration was assessed qualitatively in the internal capsule and quantitatively by measuring brainstem asymmetry. We found a strong correlation between motor dysfunction of the upper and lower limb and the lateral extent of the periventricular lesion measured on the respective semi-coronal planes. The total lesion volume and the degree of Wallerian degeneration correlated less strongly, both reaching statistical significance only with motor impairment of the hand.


9-O monofilament polydioxanone (PDS): a new synthetic absorbable suture for cataract wound closure.
This study describes the intraoperative and postoperative performance of 9-O monofilament Polydioxanone (PDS) suture for cataract wound closure. This newly developed synthetic absorbable suture is designed to meet the need for a "long-term" absorbable suture material suitable for cataract wound closure following intraocular lens implantation. Polydioxanone's advantages over currently used "short-term" absorbables and biodegradable nylon are described in this report.


Agronomic importance of first development of chickpea (Cicer arietinum L.) under semi-arid conditions: I. Effect of powder humic acid.
Due to slow growth and weakness of the first development of chickpea (Cicer arietinum L.) plant could not combatted with weeds and easily get caught up by Ascochyta blight (Ascochyta rabiei (Pass) Labr.) disease; esp. under the late sowing and semi-arid conditions, due to effect of biotic and abiotic stress factors, significant yield losses could be arised. To be able to avoid from them is only possible to accelerate the first development of this crop. So, one of the best solutions is to use of "soil conditioner" chemical compounds such as HA at optimum dose. With this aim, it was established in order to find out the optimum dose range of HA. Three doses (D0 = 0 g, D1 = 100 g and D2 = 200 g), four varieties (V1 = Er-99, V2 = Gökçe, V3 = ILC-482 and V4 = Australia) and five yield components Plant Height (PH), First pod height (FPH), Number of Branches per Plant (NBP), Number of Pods per Plant (NPP) and Number of seeds per pod (NSP) were investigated. Obtained results are: Recommended (Optimum) HA doses and interactions were ranged and found as (V4 > V2 = V3 > V1); (D2 > D0 = D1); (D1 x V4) for the PH and FPH, (D2 x V3) for the NBP and (D1 x V3) for the NPP, respectively. It was concluded that when the recommended HA dose applied, it was seen that the first development has been clearly accelerated and increased under the semi-arid conditions in terms of investigated traits and cultivars.


Analgesic efficacy of anterior iliopsoas muscle space block combined with local infiltration analgesia after total hip arthroplasty: A prospective, double-blind, placebo-controlled study.
The present study aimed to evaluate the efficacy of ultrasound-guided anterior iliopsoas muscle space block (AIMSB) combined with local infiltration analgesia (LIA) for pain management and recovery in patients who have undergone total hip arthroplasty (THA) via a posterolateral approach. In this prospective, double-blind, placebo-controlled study, 80 patients undergoing primary THA under general anesthesia were included in the final analysis between March 22, 2022, and June 1, 2022. All patients were randomly assigned to receive AIMSB combined with LIA (AIMSB group, n = 40) or sham AIMSB and LIA (Sham group, n = 40). The primary outcome was cumulative morphine consumption (mg) within 24 h after surgery. Secondary outcomes were pain scores on a visual analog scale (VAS) at rest or during motion after surgery, time to first rescue analgesia, cumulative morphine consumption during hospitalization, intraoperative consumption of opioids, postoperative recovery, and postoperative adverse effects. Patients in the AIMSB group consumed significantly less morphine than the Sham group within the first 24 h and throughout hospitalization, as well as smaller amounts of intraoperative opioids. Also, significantly lower pain scores were recorded at rest or during motion within 24 h after surgery in AIMSB patients. Patients in the AIMSB group recovered more quickly than Sham patients. No significant difference was observed in quadriceps strength and postoperative complications between the two groups. Compared to treatment with LIA alone, ultrasound-guided AIMSB combined with LIA can provide better postoperative pain relief, decrease opioid consumption, promote motor sparing, and enhance the recovery of THA patients.


Role of plasma adiponectin /C-reactive protein ratio in obesity and type 2 diabetes among African Americans.
Obesity is a modifiable risk factor for hypertension and T2D. Objective(s): We examined relations between fasting plasma adiponectin (ADIP), C-reactive protein (CRP) concentrations and markers of T2D in African Americans (AA). Fasting plasma ADIP, CRP, Insulin (IN), HOMA-IR, lipid profiles, body fat percent (%BF), waist circumference (WC), body mass index (BMI) and blood pressure measures were determined in AA women (W: n=77) and men (M: n=34). Participants were classified into: 1) Normal fasting glucose (FG) and Normal %BF; 2) Normal FG and High %BF; and 3) High FG. Compared to men, women had significantly higher mean ADIP (W: 31.4±2.9 vs. M: 18.0±4.4 ng/L), CRP (W: 3.2±0.3 vs. M: 2.0±0.5 mg/L), %BF (W: 41.2±0.9 vs. M: 27.2±1.3), and BMI (W: 32.3±0.7 vs. M: 29.2±1.1 kg/m2). Women with normal FG and %BF had significantly higher ADIP (64.0±6.0) and lower CRP (1.3±0.6) concentrations than normal FG/ high %BF (ADIP: 37.0±5.0 and CRP: 3.1 ±0.5) and high FG (ADIP: 15.1±4.1 and CRP: 4.0 ± 0.5) groups. Women with high ADIP to CRP ratio had favorable metabolic and anthropometric profiles. Low ADIP and high CRP are associated with excessive %BF and FG in AA women. ADIP/CRP, may be useful for detecting metabolic dysregulation.


Molecular cloning of macrophin, a human homologue of Drosophila kakapo with a close structural similarity to plectin and dystrophin.
We have determined the complete cDNA coding sequence of a novel cytoskeletal protein by the degenerative primer-mediated PCR strategy. This novel gene, named as macrophin (microfilament and actin filament cross-linker protein related to plectin and dystrophin, Accession No. AB029290), appears to be a human homologue of a Drosophila gene, kakapo, and shows close similarity to plectin and dystrophin on the search of BLAST homology-computed database. Comparison of the deduced protein sequences for macrophin and kakapo revealed that they were 66% similar, and both of them have an NH2-terminal actin-binding domain, a central rod region composed of spectrin-like repeats, and COOH-terminal Gas2-related region. The predicted sequences for macrophin are 5430 amino acids in length with a calculated molecular mass of 620 kDa, which is one of the largest size idenfied in human cytoskeletal proteins. High expression of macrophin was observed in brain, heart, lung, placenta, liver, kidney, and pancreas. In pancreas, we found that macrophin was specifically expressed in acinar cells than islet cells by in situ hybridization. By using radiation hybrid panel, we have mapped the macrophin gene to the chromosome 1p31-32.


Measuring Change in Symptoms of Neurobehavioural Disability: Responsiveness of the St Andrew's-Swansea Neurobehavioural Outcome Scale.
Neurobehavioural disability (NBD) after acquired brain injury (ABI) is often associated with poor outcome. The "St Andrew's-Swansea Neurobehavioural Outcome Scale" (SASNOS) was developed to measure NBD in a range of applications. Two of the "holy trinity" of psychometric properties, reliability and validity, have been comprehensively mapped, but the extent to which SASNOS meets the third, responsiveness, has not been investigated. Demonstrating responsiveness is essential in instruments employed in repeated measurement scenarios to confirm their ability to discriminate real change from error. However, there is no single agreed method for determining responsiveness. For some instruments, this property remains unexplored. A difference in scores attaining statistical significance for aggregate data is frequently cited as support for this construct, but this approach remains heavily criticized. This study explores responsiveness of SASNOS. Consecutive SASNOS assessments completed over varying times for 145 individuals participating in neurobehavioural rehabilitation, drawn from multiple services, were compiled into a retrospective sample of convenience. Multiple methods were employed to confirm internal responsiveness, including those identifying statistically significant change, minimally detectable change and minimally important change. All methods confirmed responsiveness as a psychometric property of SASNOS; the extent depended on method used and NBD domain investigated. A number of indicators are presented, which equip clinicians and researchers with options to interpret results from repeated assessments, including the individual level in the context of rehabilitation. SASNOS reliably measures change over time in NBD symptoms, further confirming its suitability as an instrument for investigating multidimensional outcomes of ABI.


Single-cell RNA sequencing of mouse brain and lung vascular and vessel-associated cell types.
Vascular diseases are major causes of death, yet our understanding of the cellular constituents of blood vessels, including how differences in their gene expression profiles create diversity in vascular structure and function, is limited. In this paper, we describe a single-cell RNA sequencing (scRNA-seq) dataset that defines vascular and vessel-associated cell types and subtypes in mouse brain and lung. The dataset contains 3,436 single cell transcriptomes from mouse brain, which formed 15 distinct clusters corresponding to cell (sub)types, and another 1,504 single cell transcriptomes from mouse lung, which formed 17 cell clusters. In order to allow user-friendly access to our data, we constructed a searchable database (http://betsholtzlab.org/VascularSingleCells/database.html). Our dataset constitutes a comprehensive molecular atlas of vascular and vessel-associated cell types in the mouse brain and lung, and as such provides a strong foundation for future studies of vascular development and diseases.


Systematic Numerical Analysis of Magnetic Field Partial Body Exposure and Comparison With Occupational Exposure Limit Values According to European Directive 2013/35/EU.
According to European Directive 2013/35/EU, exposure limit values for protection against sensory and health effects are met if external electric or magnetic fields are below corresponding low and high action levels for head, trunk and limb exposure, respectively. In order to verify this assumption even for more realistic exposure situations deviating from strictly uniform fields, systematic numerical computations using anatomical body models exposed to worst-case magnetic field gradients in different body regions are conducted. Applying magnetic fields according to low action level to the head and high action level to trunk and limbs at the same time results in slightly increased in situ electric field strengths in central nervous system tissues, compared to uniform exposure at the low action level. The extent of this increase depends on field orientation, field gradient (slope from low to high action level), and relative position between body region and field gradient. While this increase does not lead to conflicts with the exposure limit values for frequencies of 50 Hz and higher, violation of corresponding exposure limit values could be observed at 16.67 Hz.


Epigenetic modification contributes to the expression divergence of three TaEXPA1 homoeologs in hexaploid wheat (Triticum aestivum).
Common wheat is a hexaploid species with most of the genes present as triplicate homoeologs. Expression divergences of homoeologs are frequently observed in wheat, as well as in other polyploid plants. However, the mechanisms underlying this phenomenon are poorly understood. Expansin genes play important roles in the regulation of cell size, as well as organ size. We found that all three TaEXPA1 homoeologs were silenced in seedling roots. In seedling leaves, TaEXPA1-A and TaEXPA1-D were expressed, but TaEXPA1-B was silenced. Further analysis revealed that silencing of TaEXPA1-B in leaves occurred after the formation of the hexaploid. Chromatin immunoprecipitation assays revealed that the transcriptional silencing of three TaEXPA1 homoeologs in roots was correlated with an increased level of H3K9 dimethylation and decreased levels of H3K4 trimethylation and H3K9 acetylation. Reactivation of TaEXPA1-A and TaEXPA1-D expression in leaves was correlated with increased levels of H3K4 trimethylation and H3K9 acetylation, and decreased levels of H3K9 dimethylation in their promoters, respectively. Moreover, a higher level of cytosine methylation was detected in the promoter region of TaEXPA1-B, which may contribute to its silencing in leaves. We demonstrated that epigenetic modifications contribute to the expression divergence of three TaEXPA1 homoeologs during wheat development.


A Novel Negative Allosteric Modulator Selective for GluN2C/2D-Containing NMDA Receptors Inhibits Synaptic Transmission in Hippocampal Interneurons.
N-Methyl-d-aspartate receptors (NMDARs) are ionotropic glutamate receptors that mediate excitatory synaptic transmission and have been implicated in numerous neurological disorders. NMDARs typically comprise two GluN1 and two GluN2 subunits. The four GluN2 subtypes (GluN2A-GluN2D) have distinct functional properties and gene expression patterns, which contribute to diverse functional roles for NMDARs in the brain. Here, we present a series of GluN2C/2D-selective negative allosteric modulators built around a N-aryl benzamide (NAB) core. The prototypical compound, NAB-14, is >800-fold selective for recombinant GluN2C/GluN2D over GluN2A/GluN2B in Xenopus oocytes and has an IC50 value of 580 nM at recombinant GluN2D-containing receptors expressed in mammalian cells. NAB-14 inhibits triheteromeric (GluN1/GluN2A/GluN2C) NMDARs with modestly reduced potency and efficacy compared to diheteromeric (GluN1/GluN2C/GluN2C) receptors. Site-directed mutagenesis suggests that structural determinants for NAB-14 inhibition reside in the GluN2D M1 transmembrane helix. NAB-14 inhibits GluN2D-mediated synaptic currents in rat subthalamic neurons and mouse hippocampal interneurons, but has no effect on synaptic transmission in hippocampal pyramidal neurons, which do not express GluN2C or GluN2D. This series possesses some druglike physical properties and modest brain permeability in rat and mouse. Altogether, this work identifies a new series of negative allosteric modulators that are valuable tools for studying GluN2C- and GluN2D-containing NMDAR function in brain circuits, and suggests that the series has the potential to be developed into therapies for selectively modulating brain circuits involving the GluN2C and GluN2D subunits.


Cancer Loyalty Card Study (CLOCS): feasibility outcomes for an observational case-control study focusing on the patient interval in ovarian cancer.
Ovarian cancer symptoms are often non-specific and can be normalised before patients seek medical help. The Cancer Loyalty Card Study investigated self-management behaviours of patients with ovarian cancer prior to their diagnosis using loyalty card data collected by two UK-based high street retailers. Here, we discuss the feasibility outcomes for this novel research. Observational case-control study. Control participants were invited to the study using social media and other sources from the general public. Once consented, control participants were required to submit proof of identification (ID) for their loyalty card data to be shared. Cases were identified using unique National Health Service (NHS) numbers (a proxy for ID) and were recruited through 12 NHS tertiary care clinics. Women in the UK, 18 years or older, with at least one of the participating high street retailers' loyalty cards. Those with an ovarian cancer diagnosis within 2 years of recruitment were considered cases, and those without an ovarian cancer diagnosis were considered controls. Recruitment rates, demographics of participants and identification of any barriers to recruitment. In total, 182 cases and 427 controls were recruited with significant differences by age, number of people in participants' households and the geographical region in the UK. However, only 37% (n=160/427) of control participants provided sufficient ID details and 81% (n=130/160) matched retailers' records. The majority of the participants provided complete responses to the 24-Item Ovarian Risk Questionnaire. Our findings show that recruitment to a study aiming to understand self-care behaviours using loyalty card data is challenging but feasible. The general public were willing to share their data for health research. Barriers in data sharing mechanisms need to be addressed to maximise participant retention. ISRCTN14897082, CPMS 43323, NCT03994653.


Multifunctional aptamer-based nanoparticles for targeted drug delivery to circumvent cancer resistance.
By its unique advantages over traditional medicine, nanomedicine has offered new strategies for cancer treatment. In particular, the development of drug delivery strategies has focused on nanoscale particles to improve bioavailability. However, many of these nanoparticles are unable to overcome tumor resistance to chemotherapeutic agents. Recently, new opportunities for drug delivery have been provided by oligonucleotides that can self-assemble into three-dimensional nanostructures. In this work, we have designed and developed functional DNA nanostructures to deliver the chemotherapy drug doxorubicin (Dox) to resistant cancer cells. These nanostructures have two components. The first component is a DNA aptamer, which forms a dimeric G-quadruplex nanostructure to target cancer cells by binding with nucleolin. The second component is double-stranded DNA (dsDNA), which is rich in -GC- base pairs that can be applied for Dox delivery. We demonstrated that Dox was able to efficiently intercalate into dsDNA and this intercalation did not affect the aptamer's three-dimensional structure. In addition, the Aptamer-dsDNA (ApS) nanoparticle showed good stability and protected the dsDNA from degradation in bovine serum. More importantly, the ApS&Dox nanoparticle efficiently reversed the resistance of human breast cancer cells to Dox. The mechanism circumventing doxorubicin resistance by ApS&Dox nanoparticles may be predominantly by cell cycle arrest in S phase, effectively increased cell uptake and decreased cell efflux of doxorubicin. Furthermore, the ApS&Dox nanoparticles could effectively inhibit tumor growth, while less cardiotoxicity was observed. Overall, this functional DNA nanostructure provides new insights into the design of nanocarriers to overcome multidrug resistance through targeted drug delivery.


Inhibition of urokinase plasminogen activator "uPA" activity alters ethanol consumption and conditioned place preference in mice.
Urokinase plasminogen activator, uPA, is a serine protease implicated in addiction to drugs of abuse. Using its specific inhibitor, B428, we and others have characterized the role of uPA in the rewarding properties of psychostimulants, including cocaine and amphetamine, but none have examined the role of uPA in ethanol use disorders. Therefore, in the current study, we extended our observations to the role of uPA in ethanol consumption and ethanol-induced conditioned place preference. The general aim of the present series of experiments was to investigate the effects of the administration of the B428 on voluntary alcohol intake and ethanol conditioned reward. A two-bottle choice, unlimited-access paradigm was used to compare ethanol intake between vehicle- and 3, 10, and 30 mg/kg B428-administered mice. For this purpose, the mice were presented with an ethanol solution (2.5%-20%) and water, at each concentration for 4 days, and their consumption was measured daily. Consumption of saccharin and quinine solutions was also measured. Systemic administration of B428 dose-dependently decreased ethanol intake and preference. Additionally, B428 mice did not differ from vehicle mice in their intake of graded solutions of tastants, suggesting that the uPA inhibition did not alter taste function. Also, ethanol metabolism was not affected following B428 injection. More importantly, 1.5 g/kg ethanol-induced conditioned place preference acquisition was blocked following B428 administration. Taken together, our results are the first to implicate uPA inhibition in the regulation of ethanol consumption and preference, and suggest that uPA may be considered as a possible therapeutic drug target for alcoholism and abstinence.


Relationship between household income and mental disorders: findings from a population-based longitudinal study.
There has been increasing concern about the impact of the global economic recession on mental health. To date, findings on the relationship between income and mental illness have been mixed. Some studies have found that lower income is associated with mental illness, while other studies have not found this relationship. To examine the relationship between income, mental disorders, and suicide attempts. Prospective, longitudinal, nationally representative survey. United States general population. A total of 34,653 noninstitutionalized adults (aged ≥20 years) interviewed at 2 time points 3 years apart. Lifetime DSM-IV Axis I and Axis II mental disorders and lifetime suicide attempts, as well as incident mental disorders and change in income during the follow-up period. After adjusting for potential confounders, the presence of most of the lifetime Axis I and Axis II mental disorders was associated with lower levels of income. Participants with household income of less than $20,000 per year were at increased risk of incident mood disorders during the 3-year follow-up period in comparison with those with income of $70,000 or more per year. A decrease in household income during the 2 time points was also associated with an increased risk of incident mood, anxiety, or substance use disorders (adjusted odds ratio, 1.30; 99% confidence interval, 1.06-1.60) in comparison with respondents with no change in income. Baseline presence of mental disorders did not increase the risk of change in personal or household income in the follow-up period. Low levels of household income are associated with several lifetime mental disorders and suicide attempts, and a reduction in household income is associated with increased risk for incident mental disorders. Policymakers need to consider optimal methods of intervention for mental disorders and suicidal behavior among low-income individuals.


Reconstruction of the transected cat spinal cord following NeuroGel implantation: axonal tracing, immunohistochemical and ultrastructural studies.
This study examined the ability of NeuroGel, a biocompatible porous poly [N-(2-hydroxypropyl) methacrylamide] hydrogel, to establish a permissive environment across a 3 mm gap in the cat spinal cord in order to promote tissue reconstitution and axonal regeneration across the lesion. Animals with NeuroGel implants were compared to transection-only controls and observed for 21 months. The hydrogel formed a stable bridge between the cord segments. Six months after reconstructive surgery, it was densely infiltrated by a reparative tissue composed of glial cells, capillary vessels and axonal fibres. Axonal labelling and double immunostaining for neurofilaments and myelin basic protein, showed that descending supraspinal axons of the ventral funiculus and afferent fibres of the dorsal column regenerated across the reconstructed lesion. Fifteen months after reconstructive surgery, axons had grown, at least, 12 mm into the distal cord tissue, and in the rostral cord there was labelling of neurons of the intermediate gray matter. Electron microscopy showed that after 9 months, most of the regenerating axons were myelinated, principally by Schwann cells. Newly formed neurons presumably from precursor cells of the ependyma and/or migrating neurons were observed within the reparative tissue after 21 months. Results indicate that functional deficit, as assessed by treadmill training, and morphological changes following double transection of the spinal cord can be modified by the implantation of NeuroGel. This technology offers the potential to promote the formation of a neural tissue equivalent via a reparative neohistogenesis process, that facilitates and supports regenerative growth of axons.


Secretory Peptides as Bullets: Effector Peptides from Pathogens against Antimicrobial Peptides from Soybean.
Soybean is an important crop as both human food and animal feed. However, the yield of soybean is heavily impacted by biotic stresses including insect attack and pathogen infection. Insect bites usually make the plants vulnerable to pathogen infection, which causes diseases. Fungi, oomycetes, bacteria, viruses, and nematodes are major soybean pathogens. The infection by pathogens and the defenses mounted by soybean are an interactive and dynamic process. Using fungi, oomycetes, and bacteria as examples, we will discuss the recognition of pathogens by soybean at the molecular level. In this review, we will discuss both the secretory peptides for soybean plant infection and those for pathogen inhibition. Pathogenic secretory peptides and peptides secreted by soybean and its associated microbes will be included. We will also explore the possible use of externally applied antimicrobial peptides identical to those secreted by soybean and its associated microbes as biopesticides.


Effects of pharmacological manipulation of the kappa opioid receptors on the aversive effects of nicotine.
Nicotine, an addictive component of tobacco smoke, produces both rewarding and aversive effects. Increasing the aversive effects of nicotine may help in promoting smoking cessation. However, neural targets mediating the aversive effects of nicotine have not been fully identified. In this study, we evaluated the role of kappa opioid receptors (KORs) in the aversive effects of nicotine (0.4 mg/kg, base; s.c.) using the nicotine-induced conditioned taste aversion (CTA) model in Wistar rats. The KORs were activated using the selective KOR agonist (±)U-50,488H (0, 0.03, 0.15 & 0.3mg/kg; s.c.) and inhibited using the KOR antagonist nor-binaltorphimine (nor-BNI; 0, 15 & 30mg/kg; s.c.) in separate groups of rats using a between-subjects design. Pretreatment with the KOR agonist (±)U-50,488H (0.3mg/kg) significantly increased aversion for the nicotine-associated solution. Additionally, (±)U-50,488H (0.3mg/kg) on its own induced aversion to the flavored solution associated with it even in the absence of nicotine, suggesting that the KOR agonist induced increase in nicotine-induced aversion was an additive effect. Interestingly, administration of the KOR antagonist nor-BNI (30mg/kg) prior to conditioning with nicotine/saline, but not after conditioning with nicotine/saline, attenuated nicotine-induced aversive effects compared to saline controls. Taken together, these data suggest a role for KORs in the aversive effects of nicotine.


Proteome-wide structural analysis quantifies structural conservation across distant species.
Traditional evolutionary biology research mainly relies on sequence information to infer evolutionary relationships between genes or proteins. In contrast, protein structural information has long been overlooked, although structures are more conserved and closely linked to the functions than the sequences. To address this gap, we conducted a proteome-wide structural analysis using experimental and computed protein structures for organisms from the three distinct domains, including Homo sapiens (eukarya), Escherichia coli (bacteria), and Methanocaldococcus jannaschii (archaea). We reveal the distribution of structural similarity and sequence identity at the genomic level and characterize the twilight zone, where signals obtained from sequence alignment are blurred and evolutionary relationships cannot be inferred unambiguously. We find that structurally similar homologous protein pairs in the twilight zone account for ∼0.004%-0.021% of all possible protein pair combinations, which translates to ∼8%-32% of the protein-coding genes, depending on the species under comparison. In addition, by comparing the structural homologs, we show that human proteins involved in the energy supply are more similar to their E. coli homologs, whereas proteins relating to the central dogma are more similar to their M. jannaschii homologs. We also identify a bacterial GPCR homolog in the E. coli proteome that displays distinctive domain architecture. Our results shed light on the characteristics of the twilight zone and the origin of different pathways from a protein structure perspective, highlighting an exciting new frontier in evolutionary biology.


Willingness to Adopt mHealth Among Chinese Parents During the COVID-19 Outbreak: Cross-sectional Questionnaire Study.
Parental involvement in mobile health (mHealth) to consult with medical professionals appears to be prevalent in China with the rapid development of the internet. More parents with busy jobs have chosen to use mHealth. During the ongoing COVID-19 outbreak, mHealth can assist with health promotion, directions for medication use, and disease diagnosis via online chat and video consultation without contacting others. To our knowledge, no studies have been performed to explore the role of mHealth in parents' attitudes toward child health care at home during the COVID-19 outbreak. This study aims to identify the associated factors of willingness to adopt mHealth among Chinese parents during the COVID-19 outbreak and to explore the correlation between the frequency of adopting mHealth and parents' attitudes toward child health care at home. Chinese parents were asked to complete an online survey from January 25 to February 15, 2020. The questionnaire comprised of two parts with a total of 16 items, including parents' demographic variables and attitudes toward child health care at home. By multivariate logistic regression, we explored factors associated with parents' willingness to adopt mHealth during the COVID-19 outbreak. Pearson chi-square tests were used to reveal the correlation between the frequency of adopting mHealth and parents' attitudes toward child health care at home. A total of 254 parents enrolled, and 202 (79.5%) parents were willing to adopt mHealth during the COVID-19 outbreak. Parents' age (26-35 years: adjusted odds ratio [AOR] 8.114, 95% CI 1.471-44.764), parents' interest in the COVID-19 pandemic (moderate: AOR 8.753, 95% CI 2.009-38.127; high: AOR 22.194, 95% CI 5.509-89.411), the source that recommended mHealth (medical health providers: AOR 4.257, 95% CI 1.439-12.596), the presence of chronic disease in their children (yes: AOR 20.844, 95% CI 4.600-94.443), parents' duration of daily internet use (4-6 hours: AOR 6.487, 95% CI 1.870-22.495; >6 hours: AOR 8.766, 95% CI 1.883-40.804), and adoption of mHealth before the COVID-19 outbreak (yes: AOR 3.413, 95% CI 1.234-9.444) were significantly correlated with the parents' willingness to adopt mHealth during the COVID-19 outbreak. The frequency of mHealth use among parents was correlated with their behaviors in regard to handwashing (χ26=18.967, P=.004), mask wearing (χ26=45.364, P<.001), frequency of leaving the home (χ26=16.767, P=.01), room disinfection and ventilation (χ26=19.515, P=.003), temperature checking (χ26=17.47, P=.007), and mental health care of children (χ26=63.810, P<.001) during the COVID-19 pandemic. We found various objective factors that were associated with parents' willingness to adopt mHealth during the COVID-19 outbreak. Overall, parents' willingness to adopt mHealth was high. The frequency of mHealth use among parents was correlated with their attitudes toward child health care at home. The option of mHealth to patients at home during the COVID-19 outbreak would be beneficial for education and improvement in self-management of child health care at home.


Theory-Based Digital Interventions to Improve Asthma Self-Management Outcomes: Systematic Review.
Asthma is a chronic disease requiring effective self-management to control it and prevent mortality. The use of theory-informed digital interventions promoting asthma self-management is increasing. However, there is limited knowledge concerning how and to what extent psychological theory has been applied to the development of digital interventions, or how using theory impacts outcomes. The study aimed to examine the use and application of theory in the development of digital interventions to enhance asthma self-management and to evaluate the effectiveness of theory-based interventions in improving adherence, self-management, and clinical outcomes. Electronic databases (CENTRAL, MEDLINE, EMBASE, and PsycINFO) were searched systematically using predetermined terms. Additional studies were identified by scanning references within relevant studies. Two researchers screened titles and abstracts against predefined inclusion criteria; a third resolved discrepancies. Full-text review was undertaken for relevant studies. Those meeting inclusion criteria were assessed for risk of bias using the Cochrane Collaboration tool. The review was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement. Study outcomes were classified as medication adherence, self-management, asthma control, clinical markers of health, quality of life, other quality of life outcomes, and health care utilization. Effectiveness was calculated as an average outcome score based on the study's reported significance. The Theory Coding Scheme (TCS) was used to establish the extent to which each intervention had applied theory and which theoretical constructs or behavioral determinants were addressed. Associations between TCS scores and asthma outcomes were described within a narrative synthesis. Fourteen studies evaluating 14 different digital interventions were included in this review. The most commonly cited theories were Social Cognitive Theory, Health Belief Model, and Self-Efficacy Theory. A greater use of theory in the development of interventions was correlated with effective outcomes (r=.657; P=.01): only the 3 studies that met >60% of the different uses of theory assessed by the TCS were effective on all behavioral and clinical outcomes measured. None of the 11 studies that met ≤60% of the TCS criteria were fully effective; however, 3 interventions were partially effective (ie, the intervention had a significant impact on some, but not all, of the outcomes measured). Most studies lacked detail on the theoretical constructs and how they were applied to the development and application of the intervention. These findings suggest that greater use of theory in the development and application of digital self-management interventions for asthma may increase their effectiveness. The application of theory alone may not be enough to yield a successful intervention, and other factors (eg, the context in which the intervention is used) should be considered. A systematic approach to the use of theory to guide the design, selection, and application of intervention techniques is needed.


Regioselective oxidative coupling of 4-hydroxystilbenes: synthesis of resveratrol and epsilon-viniferin (E)-dehydrodimers.
Treatment of 5-[2-(4-hydroxyphenyl)vinyl]benzene-1,3-diol (resveratrol) with an equimolar amount of silver(I) acetate in dry MeOH at 50 degrees C for 1 h followed by chromatographic purification with a short silica gel column allowed the isolation of its (E)-dehydrodimer, 5-[5-[2-(3,5-dihydroxyphenyl)vinyl]-2-(4-hydroxyphenyl)-2,3-dihydrobenzofuran-3-yl]benzene-1,3-diol, as a racemic mixture in high yield. The present method was applicable to the oxidative dimerization of 4-hydroxystilbenes such as trans-styrylphenol and 5-[6-hydroxy-2-(4-hydroxyphenyl)-4-[2-(4-hydroxyphenyl)vinyl]-2,3-dihydrobenzofuran-3-yl]benzene-1,3-diol (epsilon-viniferin) leading to the corresponding 2-(4-hydroxyphenyl)-2,3-dihydrobenzofurans possessing various types of biological activities.


Does paclitaxel improve the chemoradiotherapy of locoregionally advanced esophageal cancer? A nonrandomized comparison with fluorouracil-based therapy.
A phase II trial of accelerated fractionation radiation with concurrent cisplatin and paclitaxel chemotherapy was performed to investigate the role of the paclitaxel, when substituted for fluorouracil (5-FU), in the chemoradiotherapy of esophageal cancer. Patients with an esophageal ultrasound stage of T(3) or N(1) or M(1) (nodal) esophageal cancer were treated with two courses of a cisplatin infusion (20 mg/m(2)/d for 4 days) and paclitaxel (175 mg/m(2) over 24 hours) concurrent with a split course of accelerated fractionation radiation (1.5 Gy bid to a total dose of 45 Gy). Surgical resection was performed 4 to 6 weeks later followed by a single identical postoperative course of chemoradiotherapy (24 Gy) in patients with significant residual tumor at surgery. Toxicity and results of this treatment were retrospectively compared with our previous 5-FU and cisplatin chemoradiotherapy experience. Between September 1995 and July 1997, 40 patients were entered onto this study. Although dysphagia proved worse in our 5-FU-treated patients, profound leukopenia and a need for unplanned hospitalization were significantly more common in the paclitaxel group. Thirty-seven patients (93%) proved resectable for cure. The 3-year projected overall survival is 30%, locoregional control is 81%, and distant metastatic disease control is 44%. When compared with a similarly staged cohort of 5-FU-treated patients, there was no advantage for any survival function studied. This paclitaxel-based treatment regimen for locoregionally advanced esophageal cancer produced increased toxicity with no improvement in results when compared with our previous 5-FU experience. Paclitaxel-based treatments must be carefully and prospectively studied before their incorporation into the standard management of esophageal cancer.


Inter- and intraday variability in major electrocardiogram intervals and amplitudes in healthy men and women.
The ECG may vary during the day (intra-day), and between days (interday), for the same subject. Variability in ECG characteristic measurements between different investigators is well documented and is often large. During days 1-6 of each placebo period of a two-way crossover Phase I study, digital ECGs were recorded at about 8 and 12 AM in 16 healthy volunteers (8 men, 8 women). Two observers independently analyzed leads V2 and V6 using EClysis software. The durations and amplitudes of major ECG waves and the intervals between major electrocardiographic events were analyzed in a mixed model ANOVA, in which subject, observer, time, and day were treated as random factors. The influence of various corrections for heart rate on the variability of QT intervals was investigated. The difference among subjects explained between 44-81% of the total variability in ECG intervals and amplitudes. Overall, inter- and intraday variability was not statistically significant for any variable. The individualized exponential correction of the QT interval for heart rate eliminated the QT interval dependence on the RR interval in all subjects. Changes in T wave morphology and shortening of the QT interval from morning to noon were observed in ten subjects. The interobserver variability was close to zero (SD < 0.005 ms) for all variables except the PQ interval (SD 1.4 ms). The various sources of variability in determinations of ECG wave characteristics should be considered in the design of clinical studies. The use of EClysis software for ECG measurements is this study made the results highly observer independent.


Genome-wide identification and expression analysis of SWI1 genes in Boechera species.
As a mode of reproduction in plants, apomixis leads to the generation of clones via seeds. Apomictic plants form viable diploid female gametes without meiosis (apomeiosis) and produce embryos without fertilization (parthenogenesis). Apomeiosis, as a major component of apomixis, has recently been reported in some Arabidopsis thaliana mutants; dyad mutants of SWI1 showed developmental processes common to apomeiosis, such as producing functional diploid gametes. However, the orthologs of SWI1 genes in natural apomicts has not been previously reported. To identify the relationship between the SWI1 gene and the apomeiosis process, we isolated and sequenced SWI1 orthologs from Boechera species, including apomictic and sexual species. Boechera species are close relatives of A. thaliana and thus are advantageous model species for apomixis research. The SWI1 cDNAs were obtained by RT-PCR from apomictic and sexual Boechera young flower buds. We sequenced partial SWI1 transcripts that were 650bp for B. holboellii and 684bp for B. stricta. These SWI1-like sequences showed 86% similarity for B. holboellii and 92% for B. stricta to the A. thaliana SWI1 transcript. We also used available genome data and amplified genomic sequences for SWI1 orthologs in B. holboellii and B. stricta. The predicted proteins contain a phospholipase C domain and a nuclear localization signal. Sequence analysis did not show significant mutations related to apomixis, and phylogenetic analysis showed that SWI1-like sequences were common across plant families, regardless of the presence of a sexual or apomictic reproduction system. We also investigated the expression levels of SWI1 mRNA in the B. holboellii and B. stricta young unopened flower buds and found that relatively high levels of expression occurred in apomicts.


Expression of cyclins A, E and topoisomerase II alpha correlates with centrosome amplification and genomic instability and influences the reliability of cytometric S-phase determination.
The progression of normal cells through the cell cycle is meticulously regulated by checkpoints guaranteeing the exact replication of the genome during S-phase and its equal division at mitosis. A prerequisite for this achievement is synchronized DNA-replication and centrosome duplication. In this context the expression of cyclins A and E has been shown to play a principal role. Our results demonstrated a correlation between centrosome amplification, cell cycle fidelity and the level of mRNA and protein expression of cyclins A and E during the part of the cell cycle defined as G1-phase by means of DNA content based histogram analysis. It is shown that the normal diploid breast cell line HTB-125, the genomically relatively stable aneuploid breast cancer cell line MCF-7, and the genomically unstable aneuploid breast cancer cell line MDA-231 differ remarkably concerning both mRNA and protein expression of the two cyclins during G1-phase. In MDA-231 cells the expression of e.g. cyclin A mRNA was found to be ten times higher than in MCF-7 cells and about 500 times higher than in HTB-125 cells. Topoisomerase II alpha showed high mRNA expression in MDA compared to MCF-7 cells, but the difference in protein expression was small. Furthermore, we measured centrosome aberrations in 8.4% of the MDA-231 cells, and in only 1.3% of the more stable aneuploid cell line MCF-7. MDA cells showed 27% more incorporation of BrdU than reflected by S-phase determination with flow cytometric DNA content analysis, whereas these values were found to be of the same size in both HTB-125 and MCF-7 cells. Our data indicate that the breast cancer cell lines MCF-7 and MDA-231, although both DNA-aneuploid, differ significantly regarding the degree of cell cycle disturbance and centrosome aberrations, which partly could explain the different genomic stability of the two cell lines. The results also question the reliability of cytometric DNA content based S-phase determination in genomically unstable tumor cell populations.


Polyionic charge density plays a key role in differential recognition of mobile ions by biopolymers.
We address the question of what are the molecular mechanisms providing discrimination between seemingly similar counterions binding to various biomolecular surfaces. In the case of protein association with Na (+) and K (+) ions, recent works proposed that specificity of carboxylate functional groups interacting with these mobile ions rationalizes the observed ionic discrimination. We probe in this work whether similar arguments may be used to explain higher propensity of Na (+) ions to associate with DNA compared with K (+) ions, which was suggested by our simulations and some experiments. By comparing our extensive molecular dynamics simulations of Na (+) and K (+) distributions around a 16-base-pair DNA oligomer, [(CGAGGTTTAAACCTCG)] 2, with additional simulations where DNA is replaced by a "soup" of monomers (dimethylphosphate anion), we conclude that DNA specificity toward Na (+)/K (+) is not determined by the underlying functional group specificity. Instead, the collective effect of DNA charges drives larger Na (+) association. To gain additional microscopic insights into the mechanisms of specificity on ionic associations in these systems, we carried out energetic analysis of the association between Na (+) and K (+) with chloride and dimethylphosphate anions. The insights gained from our computational work shed light on a number of experiments on electrolyte solutions of monovalent salts and DNA.


Caloric consequences of sugar solutions: a failure to obtain gustatory learning.
Rats decrease total caloric intake, sometimes to the point of starvation, when sugar solutions are offered in conjunction with a daily pellet meal. This phenomenon appears to be based on a tendency to behave as if overestimating the caloric value of simple sugars. In these experiments, rats were exposed to a variety of conditions that allowed the opportunity to monitor the taste and postingestive properties of sucrose or glucose solutions. Despite these repeated opportunities to learn the true caloric value of the solutions, the rats continued to exhibit the exaggerated response. The results suggest that the gustatory receptors that have evolved to detect the normally low concentrations of sugars in the natural environment may be inherently inaccurate in response to relatively high concentrations of pure sugars. Furthermore, the rat may be contraprepared to form the quantitative gustatory associations that would be necessary to correct this error tendency.


Subjective and objective variation of the tear film pre- and post-sleep.
To date, few studies have correlated the overnight effects of the preocular tear film (POTF) with subjective symptoms. This study investigates the POTF volume and stability, bulbar hyperemia (BH), tear ferning (TF) and the participant's subjective symptoms, pre- and post-sleep. Thirty subjects were recruited, consisting of two evenly distributed groups who were symptomatic of dry eye (DE) and those that were asymptomatic dry eye, determined using the McMonnies questionnaire. Subjects were evaluated at 10 p.m. (baseline), on waking at 7 a.m., and then hourly until 10 a.m. At each visit, tear meniscus height (TMH), various subjective factors, BH and POTF stability by non-invasive break-up time (NIBUT) were assessed. Tear collection was performed at 10 p.m, 7 and 10 a.m. for TF analysis. With the exception of burning, all other symptoms (comfort, dryness, clarity of vision, and grittiness) revealed an overnight change (p < 0.05) within each group, but not between the two groups (p > 0.05). Both the tear meniscus height and BH were elevated upon waking and differed significantly between test times for each group (p < 0.05), but not between groups (p > 0.05). NIBUT was lower for the DE group (p < 0.001). The non-dry eye (NDE) group did not significantly alter over time (p > 0.05), but the DE group did (p = 0.004), with a longer NIBUT in the morning. TF demonstrated a degraded pattern upon waking for both groups (p < 0.05). Most of the changes returned to baseline within an hour after waking. The properties of the POTF undergo a change during extended periods of eye closure and the human POTF is different upon waking to that present immediately before sleep. Most of the parameters determined rapidly revert to baseline levels once the POTF is allowed to refresh.


Deletion of a kinesin I motor unmasks a mechanism of homeostatic branching control by neurotrophin-3.
Development and function of highly polarized cells such as neurons depend on microtubule-associated intracellular transport, but little is known about contributions of specific molecular motors to the establishment of synaptic connections. In this study, we investigated the function of the Kinesin I heavy chain Kif5aa during retinotectal circuit formation in zebrafish. Targeted disruption of Kif5aa does not affect retinal ganglion cell differentiation, and retinal axons reach their topographically correct targets in the tectum, albeit with a delay. In vivo dynamic imaging showed that anterograde transport of mitochondria is impaired, as is synaptic transmission. Strikingly, disruption of presynaptic activity elicits upregulation of Neurotrophin-3 (Ntf3) in postsynaptic tectal cells. This in turn promotes exuberant branching of retinal axons by signaling through the TrkC receptor (Ntrk3). Thus, our study has uncovered an activity-dependent, retrograde signaling pathway that homeostatically controls axonal branching.


[Acute early postoperative intestinal obstruction in patients with gastroduodenal ulcer].
Results of surgical treatment of 4389 patients with gastroduodenal ulcer were analysed. Relaparotomy (RL) was performed in 215(4.9%) of patients for postoperative complications occurred, including 78(36.3%)--for an acute early postoperative ileus (AEPI). RL was performed in 44.3% of patients after perforated gastroduodenal ulcer suturing in 34.3%--after gastric resection, in 28.8%--after organ preserving operations. Main measures providing the effectiveness of surgical aid rendering to the AEPI patients are an early diagnosis and timely RL performance.


Low Referral Rate for Genetic Testing in Racially and Ethnically Diverse Patients Despite Universal Colorectal Cancer Screening.
Guidelines recommend that all colorectal tumors be assessed for mismatch repair deficiency, which could increase identification of patients with Lynch syndrome. This is of particular importance for minority populations, in whom hereditary syndromes are under diagnosed. We compared rates and outcomes of testing all tumor samples (universal testing) collected from a racially and ethnically diverse population for features of Lynch syndrome. We performed a retrospective analysis of colorectal tumors tested from 2012 through 2016 at 4 academic centers. Tumor samples were collected from 767 patients with colorectal cancer (52% non-Hispanic white [NHW], 26% African American, and 17% Hispanic patients). We assessed rates of tumor testing, recommendations for genetic evaluation, rates of attending a genetic evaluation, and performance of germline testing overall and by race/ethnicity. We performed univariate and multivariate regression analyses. Overall, 92% of colorectal tumors were analyzed for mismatch repair deficiency without significant differences among races/ethnicities. However, minority patients were significantly less likely to be referred for genetic evaluation (21.2% for NHW patients vs 16.9% for African American patients and 10.9% for Hispanic patients; P = .02). Rates of genetic testing were also lower among minority patients (10.7% for NHW patients vs 6.0% for AA patients and 3.1% for Hispanic patients; P < .01). On multivariate analysis, African American race, older age, and medical center were independently associated with lack of referral for genetic evaluation and genetic testing. In a retrospective analysis, we found that despite similar rates of colorectal tumor analysis, minority patients are less likely to be recommended for genetic evaluation or to undergo germline testing for Lynch syndrome. Improvements in institutional practices in follow up after tumor testing could reduce barriers to diagnosis of Lynch diagnosis in minorities.


Unique characteristics of motor adaptation during walking in young children.
Children show precocious ability in the learning of languages; is this the case with motor learning? We used split-belt walking to probe motor adaptation (a form of motor learning) in children. Data from 27 children (ages 8-36 mo) were compared with those from 10 adults. Children walked with the treadmill belts at the same speed (tied belt), followed by walking with the belts moving at different speeds (split belt) for 8-10 min, followed again by tied-belt walking (postsplit). Initial asymmetries in temporal coordination (i.e., double support time) induced by split-belt walking were slowly reduced, with most children showing an aftereffect (i.e., asymmetry in the opposite direction to the initial) in the early postsplit period, indicative of learning. In contrast, asymmetries in spatial coordination (i.e., center of oscillation) persisted during split-belt walking and no aftereffect was seen. Step length, a measure of both spatial and temporal coordination, showed intermediate effects. The time course of learning in double support and step length was slower in children than in adults. Moreover, there was a significant negative correlation between the size of the initial asymmetry during early split-belt walking (called error) and the aftereffect for step length. Hence, children may have more difficulty learning when the errors are large. The findings further suggest that the mechanisms controlling temporal and spatial adaptation are different and mature at different times.


Outbreak off canine distemper in vaccinated dogs in Finland.
Canine distemper reappeared in dogs in Finland in 1990 after a 16-year absence. In 1994 to 1995 an outbreak occurred in areas with a high density dog population which involved dogs vaccinated against distemper. The estimated total number of cases was at least 5000, and 865 cases were confirmed by indirect fluorescent antibody testing of 3649 epithelial cell samples. The signs recorded by veterinary clinicians ranged from conjunctivitis, pyrexia and anorexia to signs of respiratory and gastrointestinal illness, with an estimated mortality of 30 per cent. Of the confirmed cases 631 (73 per cent) were between three and 24 months of age; 487 of these had been vaccinated at least once and 351 (41 per cent) had a complete vaccination history. Of these 351 fully vaccinated animals the proportion of dogs vaccinated with the most popular vaccine was significantly higher than would have been expected by its market share. In total, 4676 serum samples were collected from healthy vaccinated dogs during the peak and decline of the outbreak and tested for the presence of virus neutralising antibodies. The decrease in the proportion of young dogs with antibody titres < 1/8 coincided with the decline and end of the outbreak during the spring and summer of 1995. It was concluded that a critical decrease in the population's immunity during 1990 to 1994 was a major reason for the outbreak in the summer of 1994 and that the ultimate test for vaccines is an outbreak of disease.


Reasons for Staying as a Participant in the Environmental Determinants of Diabetes in the Young (TEDDY) Longitudinal Study.
To assess parents' opinions about their participation in the longitudinal, multicenter study - The Environmental Determinants of Diabetes in the Young (TEDDY) consortium. A survey was given to parents who had been in the study for ≥ 1 year. Parents rated the importance of different reasons for staying in TEDDY and how well different study components were working. Parents were also asked if they had suggestions for making TEDDY better and if they ever had thought of leaving TEDDY and if so, why. Out of the 3336 eligible families, 2000 completed the survey (59.1%); most (77.6%) were mothers. Survey completion was more common in European than US TEDDY sites and was associated with greater maternal education, more accurate perceptions about their child's risk of type 1 diabetes, longer participation in TEDDY and excellent attendance at TEDDY visits. "Having someone watching the child for development of T1DM" was most important reason given for staying in the study; other important reasons included "Helping science discover causes of diabetes" and "Getting child's antibody results". Most parents were very satisfied with the different components of TEDDY and had not thought of leaving the study. A minority (24%) of parents acknowledged some thoughts of leaving TEDDY and cited the blood draws, being too busy/not having enough time, the demanding protocol, and food diaries as their reasons for considering leaving. The study highlights factors important for successful implementation of demanding, longitudinal protocols. Friendly, devoted, skilled and knowledgeable staff with continuity makes the family comfortable. Keeping parents involved and informed on study progress is essential as is making procedures as smooth and painless as possible. Although the study is international the survey results were convergent across countries suggesting that the results have relevance to other similar studies to retain study participants.


Personalized tooth shape estimation from radiograph and cast.
Three-dimensional geometric information of teeth is usually needed in pre- and postoperative diagnoses of orthodontic dentistry. The computerized tomography can provide comprehensive 3-D teeth geometries. However, there is still a discussion on computed tomography (CT) as a routine in orthodontic dentistry due to radiation dose. Moreover, the CT is useless when a dentist needs to extract 3-D structures from old archive files with only radiographs and casts, where patient's teeth changed ever since. In this paper, we propose a reconstruction framework for patient-specific teeth based on an integration of 2-D radiographs and digitized casts. The reconstruction is under a template-fitting framework. The shape and orientation of teeth templates are tuned in accordance with patient's radiographs. Specially, the tooth root morphology is controlled by 2-D contours in radiographs. With ray tracing and a contour plane assumption, 2-D root contours in radiographs are projected back to 3-D space, and guide tooth root deformations. Moreover, the template's crown is deformed nonrigidly to fit digitized casts that bear patient's crown details. The system allows 3-D tooth reconstruction with patient-specific geometric details from just casts and 2-D radiographs.


Regulation of BMP-dependent chondrogenesis in early limb mesenchyme by TGFbeta signals.
In the developing axial skeleton, sequential sonic hedgehog (SHH) and bone morphogenetic protein (BMP) signals are required for specification of a chondrogenic fate in presomitic tissue. A similar paradigm is thought to operate in the limb, but the signals involved are unclear. To investigate the nature of these signals, we examined BMP action in mesenchymal populations derived from the early murine limb bud (approximately embryonic day 10.5). These populations exhibited a graded response to BMPs, in which early limb mesenchymal cells (from the distal hind limb) displayed an anti-chondrogenic response, whereas BMPs promoted chondrogenesis in more mature cell populations (from the proximal fore limb). Under these conditions, multiple Gata genes were induced by BMPs and the extent of induction correlated with BMP anti-chondrogenic activity. A screen of limb-bud-expressed ligands revealed that prior short-term exposure to transforming growth factor beta1 (TGFbeta1) ameliorated the anti-chondrogenic response to BMP. Furthermore, brief activation of the TGFbeta pathway was found to be necessary for subsequent induction of chondrogenesis by BMPs. Our findings indicate that, similar to axial skeletogenesis, induction of chondrogenesis in the appendicular skeleton is a two-step process. However, the programs differ in the transient signals driving chondrogenic responsiveness to BMPs, with SHH operating in the former and TGFbeta activation in the latter.


Role of cytoskeleton in regulating fusion of nucleoli: a study using the activated mouse oocyte model.
Although fusion of nucleoli was observed during pronuclear development of zygotes and the behavior of nucleoli in pronuclei has been suggested as an indicator of embryonic developmental potential, the mechanism for nucleolar fusion is unclear. Although both cytoskeleton and the nucleolus are important cellular entities, there are no special reports on the relationship between the two. Role of cytoskeleton in regulating fusion of nucleoli was studied using the activated mouse oocyte model. Mouse oocytes were cultured for 6 h in activating medium (Ca²⁺-free CZB medium containing 10 mM SrCl₂) supplemented with or without inhibitors for cytoskeleton or protein synthesis before pronuclear formation, nucleolar fusion, and the activity of maturation-promoting factor (MPF) were examined. Whereas treatment with microfilament inhibitor cytochalasin D or B or intermediate filament inhibitor acrylamide suppressed nucleolar fusion efficiently, treatment with microtubule inhibitor demecolcine or nocodazole or protein synthesis inhibitor cycloheximide had no effect. The cytochalasin D- or acrylamide-sensitive temporal window coincided well with the reported temporal window for nucleolar fusion in activated oocytes. Whereas a continuous incubation with demecolcine prevented pronuclear formation, pronuclei formed normally when demecolcine was excluded during the first hour of activation treatment when the MPF activity dropped dramatically. The results suggest that 1) microfilaments and intermediate filaments but not microtubules support nucleolar fusion, 2) proteins required for nucleolar fusion including microfilaments and intermediate filaments are not de novo synthesized, and 3) microtubule disruption prevents pronuclear formation by activating MPF.