Ajuba, a cytosolic LIM protein, shuttles into the nucleus and affects embryonal cell proliferation and fate decisions.
Cellular adhesive events affect cell proliferation and differentiation decisions. How cell surface events mediating adhesion transduce signals to the nucleus is not well understood. After cell-cell or cell-substratum contact, cytosolic proteins are recruited to clustered adhesion receptor complexes. One such family of cytosolic proteins found at sites of cell adhesion is the Zyxin family of LIM proteins. Here we demonstrate that the family member Ajuba was recruited to the cell surface of embryonal cells, upon aggregate formation, at sites of cell-cell contact. Ajuba contained a functional nuclear export signal and shuttled into the nucleus. Importantly, accumulation of the LIM domains of Ajuba in the nucleus of P19 embryonal cells resulted in growth inhibition and spontaneous endodermal differentiation. The differentiating effect of Ajuba mapped to the third LIM domain, whereas regulation of proliferation mapped to the first and second LIM domains. Ajuba-induced endodermal differentiation of these cells correlated with the capacity to activate c-Jun kinase and required c-Jun kinase activation. These results suggest that the cytosolic LIM protein Ajuba may provide a new mechanism to transduce signals from sites of cell adhesion to the nucleus, regulating cell growth and differentiation decisions during early development.


[Role of atypical sex cells in the histogenesis of germinative testicular tumors].
Histochemical and electron microscopic study of 23 germinative tumors of the testicle and organ tissue beyond the tumor was carried out. In 78% of the observations, normal seminiferous tubules were found to contain a special kind of cell elements, atypical sex cells (ASC), not observed in health. Ultrastructural and histochemical features of undifferentiated cells comprising seminoma and embryonal cancer were found to be similar to those of ASC and gonocytes of human embryo. The results suggest that ASC are the stem tumor cells for human germinative neoplasia which are characterized by single cytohistogenesis.


Synthesis of different-sized silver nanoparticles by simply varying reaction conditions with leaf extracts of Bauhinia variegata L.
Green synthesis of nanoparticles is one of the crucial requirements in today's climate change scenario all over the world. In view of this, leaf extract (LE) of Bauhinia variegata L. possessing strong antidiabetic and antibacterial properties has been used to synthesise silver nanoparticles (SNP) in a controlled manner. Various-sized SNP (20-120 nm) were synthesised by varying incubation temperature, silver nitrate and LE concentrations. The rate of SNP synthesis and their size increased with increase in AgNO(3) concentration up to 4 mM. With increase in LE concentration, size and aggregation of SNP was increased. The size and aggregation of SNP were also increased at temperatures above and below 40°C. This has suggested that size and dispersion of SNP can be controlled by varying reaction components and conditions. Polarity-based fractionation of B. variegata LE has suggested that only water-soluble fraction is responsible for SNP synthesis. Fourier transform infrared spectroscopy analysis revealed the attachment of polyphenolic and carbohydrate moieties to SNP. The synthesised SNPs were found stable in double distilled water, BSA and phosphate buffer (pH 7.4). On the contrary, incubation of SNP with NaCl induced aggregation. This suggests the safe use of SNP for various in vivo applications.


Nonlinear rheology of surfactant wormlike micelles bridged by telechelic polymers.
We have investigated the nonlinear rheology of a soft composite transient network made of a solution of surfactant wormlike micelles (WM) in the semidilute regime that are reversibly bridged by telechelic polymers. The samples are well described, in the linear regime, as two Maxwell fluids components blends, characterized by two markedly different characteristic times. The slow mode is mainly related to the transient network of entangled WM, and the fast mode to the network of telechelic chains. In this paper we investigate the nonlinear viscoelasticity and show that the nonlinear behavior reflects as well the behavior of two coupled networks. On one hand, stress relaxation experiments and time-resolved stress response following the application of a constant shear rate show that, in the weakly nonlinear regime, these novel composite networks stiffen. A fourfold increase of the elastic modulus with respect to the linear value is reached for strain amplitude of about 200%. This strain hardening is due to the nonlinear stretching of the telechelic polymer chains. On the other hand, the samples exhibit shear banding in the highly nonlinear regime, similarly to pure semidilute solutions of WM.


Nonbacterial thrombotic endocarditis. A neurologic perspective of clinicopathologic correlations of 99 patients.
Of 13,913 patients examined at autopsy between 1939 and 1980, the diagnosis of nonbacterial thrombotic endocarditis (NBTE) was made in 99 instances. There were 53 males and 46 females ranging in age from 4 to 89 years. Vegetations were found on the aortic valve in 39; the mitral in 37; the tricuspid in nine; and the mitral in 37; the tricuspid in nine; and the pulmonic in two. Two-valve involvement was present in 12 cases. Malignant neoplasms were found in 42 autopsies. Embolism to the brain was found in 33 cases and to other organs in 62. Coagulation abnormalities were documented in 22 cases, and a distinct picture of disseminated intravascular coagulation of thrombophlebitis elsewhere in the body should arouse suspicion of NBTE. The high incidence of multiple emboli and its association with malignant neoplasms and with a variety of cardiovascular, pulmonary, renal, and gastrointestinal disorders should provide clues for recognition of this serious disorder.


The Child Asthma Link Line: a coalition-initiated, telephone-based, care coordination intervention for childhood asthma.
Childhood asthma is a complex chronic disease that poses significant challenges regarding management, and there is evidence of disparities in care. Many medical, psychosocial, and health system factors contribute to recognized poor control of this most prevalent illness among children, with resultant excessive use of emergency departments and hospitalizations for care. Recent national guidelines emphasize the need for community-based initiatives to address these critical issues. To address health system fragmentation and impact asthma outcomes, the Philadelphia Allies Against Asthma coalition developed and implemented the Child Asthma Link Line, a telephone-based care coordination and system integration program, which has been in operation since 2001. This study evaluates the effectiveness of the Child Asthma Link Line integration model to improve asthma management by measuring utilization markers of morbidity. Medicaid Managed Care Organization claims data for 59 children who received the Link Line intervention in 2003 are compared to a matched sample of 236 children who did not receive the Link Line intervention. Children in the two study groups are ages 3 through 12 years and matched on 2003 emergency department visits, age, gender, and race/ethnicity. Primary outcome variables analyzed in this study are emergency department visits, hospitalizations, and office visit claims from the follow-up year (2004). Link Line intervention children were significantly less likely to have follow-up hospitalizations than matched sample children (p = .02). Children enrolled in the Link Line were also more likely to attend outpatient office visits in the follow-up year (p = .045). In addition, Link Line children with multiple emergency department visits in 2003 were significantly less likely to have an emergency department visit in 2004 (p = .046). This coalition-developed, telephone-based, system-level intervention had a significant impact on childhood asthma morbidity as measured by utilization endpoints of follow-up hospitalizations and emergency department visits. Telephone-based care coordination and service integration may be a viable and economic way to impact childhood asthma and other chronic diseases.


Importance of thiol groups in ligand binding to D2 dopamine receptors from brain and anterior pituitary gland.
The effects of the thiol group reagent 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) on D2 dopamine receptors have been examined in three brain regions (caudate nucleus, putamen and olfactory tubercle), and in the anterior and neurointermediate lobes of the pituitary gland. Whereas the receptors in brain were insensitive to DTNB, a dose-dependent inhibition of [3H]spiperone binding to D2 receptors in both lobes of the pituitary gland was observed. The effects in the pituitary could be reversed by subsequent treatment with dithiothreitol and could be prevented by prior occupancy of the receptor binding site with a ligand. The effect of DTNB was to decrease the number of ligand-binding sites without altering the affinity of those remaining. The results show that modification of a thiol group of the D2 dopamine receptor in the two lobes of the pituitary gland tested here significantly affects ligand binding. There is therefore a difference between the D2 dopamine receptor populations in brain and pituitary in their sensitivity to modification by DTNB, and this may imply the existence of different receptor isoforms in the two issues.


Identification of yeast cell cycle regulated genes based on genomic features.
Time-course microarray experiments have been widely used to identify cell cycle regulated genes. However, the method is not effective for lowly expressed genes and is sensitive to experimental conditions. To complement microarray experiments, we propose a computational method to predict cell cycle regulated genes based on their genomic features - transcription factor binding and motif profiles. Through integrating gene-expression data with ChIP-chip binding and putative binding sites of transcription factors, our method shows high accuracy in discriminating yeast cell cycle regulated genes from non-cell cycle regulated ones. We predict 211 novel cell cycle regulated genes. Our model rediscovers the main cell cycle transcription factors and provides new insights into the regulatory mechanisms. The model also reveals a regulatory circuit mediated by a number of key cell cycle regulators. Our model suggests that the periodical pattern of cell cycle genes is largely coded in their promoter regions, which can be captured by motif and transcription factor binding data. Cell cycle is controlled by a relatively small number of master transcription factors. The concept of genomic feature based method can be readily extended to human cell cycle process and other transcriptionally regulated processes, such as tissue-specific expression.


Conversion from mouse embryonic to extra-embryonic endoderm stem cells reveals distinct differentiation capacities of pluripotent stem cell states.
The inner cell mass of the mouse pre-implantation blastocyst comprises epiblast progenitor and primitive endoderm cells of which cognate embryonic (mESCs) or extra-embryonic (XEN) stem cell lines can be derived. Importantly, each stem cell type retains the defining properties and lineage restriction of their in vivo tissue of origin. Recently, we demonstrated that XEN-like cells arise within mESC cultures. This raises the possibility that mESCs can generate self-renewing XEN cells without the requirement for gene manipulation. We have developed a novel approach to convert mESCs to XEN cells (cXEN) using growth factors. We confirm that the downregulation of the pluripotency transcription factor Nanog and the expression of primitive endoderm-associated genes Gata6, Gata4, Sox17 and Pdgfra are necessary for cXEN cell derivation. This approach highlights an important function for Fgf4 in cXEN cell derivation. Paracrine FGF signalling compensates for the loss of endogenous Fgf4, which is necessary to exit mESC self-renewal, but not for XEN cell maintenance. Our cXEN protocol also reveals that distinct pluripotent stem cells respond uniquely to differentiation promoting signals. cXEN cells can be derived from mESCs cultured with Erk and Gsk3 inhibitors (2i), and LIF, similar to conventional mESCs. However, we find that epiblast stem cells (EpiSCs) derived from the post-implantation embryo are refractory to cXEN cell establishment, consistent with the hypothesis that EpiSCs represent a pluripotent state distinct from mESCs. In all, these findings suggest that the potential of mESCs includes the capacity to give rise to both extra-embryonic and embryonic lineages.


Probiotic Lactobacillus reuteri biofilms produce antimicrobial and anti-inflammatory factors.
Commensal-derived probiotic bacteria inhibit enteric pathogens and regulate host immune responses in the gastrointestinal tract, but studies examining specific functions of beneficial microbes in the context of biofilms have been limited in scope. Lactobacillus reuteri formed biofilms that retained functions potentially advantageous to the host including modulation of cytokine output and the production of the antimicrobial agent, reuterin. Immunomodulatory activities of biofilms were demonstrated by the abilities of specific L. reuteri strains to suppress human TNF production by LPS-activated monocytoid cells. Quantification of the antimicrobial glycerol derivative, reuterin, was assessed in order to document the antipathogenic potential of probiotic biofilms. L. reuteri biofilms differed in the quantities of reuterin secreted in this physiological state. L. reuteri biofilms secreted factors that confer specific health benefits such as immunomodulation and pathogen inhibition. Future probiotic selection strategies should consider a strain's ability to perform beneficial functions as a biofilm.


Outbreak of acute gastroenteritis caused by contamination of drinking water in a factory, the Basque Country.
On 18 September 2013, the Gipuzkoa Epidemiology Unit was notified of an outbreak of acute gastroenteritis (AGE) among employees at a domestic appliance factory. The first signs of the outbreak had emerged at the end of June and at the time of the notification 30 workers were on sick leave for gastroenteritis. Some employees had had more than one episode and the main symptoms were diarrhoea and vomiting. An investigation began to identify the causative agent, assess exposure and determine the route of transmission. Data collected by a questionnaire identified 302 episodes of AGE among 238 people affected between June and September 2013. The source of water consumed was found to be a risk factor associated with the appearance of symptoms both in the crude and the adjusted analysis: odds ratio 1.8 (0.8-4.2) and 6.4 (4.2-9.8), respectively. Microbiological analysis of stool samples and of water confirmed the presence of norovirus and rotavirus. The environmental study detected a connection between an industrial use water system and drinking water at the factory. It was concluded that the outbreak was caused by mixed viral infections, due to contamination of drinking water.


Familial resemblance for resting blood pressure with particular reference to racial differences: preliminary analyses from the HERITAGE Family Study.
Resting blood pressure in both white and black families participating in the HERITAGE Family Study was analyzed using a simple familial correlation model to assess familial influences. The two samples of black and white families were analyzed separately and together, providing an opportunity to test for heterogeneity in the familial resemblance. Maximal heritability was 46% for systolic blood pressure (SBP) and 31% for diastolic blood pressure (DBP) in the pooled sample. Noticeably higher heritabilities were found in the black sample (68% for SBP and 56% for DBP) than in the white sample (43% for SBP and 24% for DBP). The patterns of familial correlations were similar in blacks and whites, with the exception that spouse resemblance was significant in white families but not in black families. These results along with the finding that the magnitude of the familial correlations was higher in the black sample than in the white sample suggest that the effects of host and familial environmental factors differ between the races.


A diagnostic polymerase chain reaction assay for Zika virus.
Zika virus (ZIKV) is a mosquito-borne flavivirus. Infection results in a dengue-like illness with fever, headache, malaise, and a maculopapular rash. Nearly all cases are mild and self-limiting but in 2007, a large outbreak of ZIKV was reported from the island of Yap (in Micronesia, northwest of Indonesia). Singapore is already endemic for dengue, and its impact on public health and economic burden is significant. Other dengue-like infections (e.g., Chikungunya virus) are present. Yet only 10% of reported dengue cases have laboratory confirmation. The identification and control of other dengue-like, mosquito-transmitted infections is thus important for the health of Singapore's population, as well as its economy. Given that ZIKV shares the same Aedes mosquito vector with both dengue and Chikungunya, it is possible that this virus is present in Singapore and causing some of the mild dengue-like illness. A specific and sensitive one-step, reverse transcription polymerase chain reaction (RT-PCR) with an internal control (IC) was designed and tested on 88 archived samples of dengue-negative, Chikungunya-negative sera from patients presenting to our hospital with a dengue-like illness, to determine the presence of ZIKV in Singapore. The assay was specific for detection of ZIKV and displayed a lower limit of detection (LoD) of 140 copies viral RNA/reaction when tested on synthetic RNA standards prepared using pooled negative patient plasma. Of the 88 samples tested, none were positive for ZIKV RNA, however, the vast majority of these were from patients admitted to hospital and further study may be warranted in community-based environments.


Fowlpox virus in the Sudan.
A virulent fowlpox virus was isolated from a natural outbreak of the disease in the Sudan. The virus was used for experimental transmission of the disease to susceptible chickens. Scarification and intravenous methods were used to infect one-month-old chickens. Scarification induced typical pox lesions at 5-7 days postinoculation, whereas intravenous injection induced only swelling of combs or wattles, at 10-11 days postinoculation, although neutralizing antibodies against fowlpox virus were detected 2, 3, and 4 weeks postinoculation. The histopathological features of the pox lesions obtained in both natural and experimental infection were described.


Kallikrein-kinin system as the dominant mechanism to counteract hyperactive renin-angiotensin system.
The renin-angiotensin system (RAS) generates, maintains, and makes worse hypertension and cardiovascular diseases (CVDs) through its biologically active component angiotensin II (Ang II), that causes vasoconstriction, sodium retention, and structural alterations of the heart and the arteries. A few endogenous vasodilators, kinins, natriuretic peptides, and possibly angiotensin (1-7), exert opposite actions and may provide useful therapeutic agents. As endothelial autacoids, the kinins are potent vasodilators, active natriuretics, and protectors of the endothelium. Indeed, the kallikrein-kinin system (KKS) is considered the dominant mechanism for counteracting the detrimental effects of the hyperactive RAS. The 2 systems, RAS and KKS, are controlled by the angiotensin-converting enzyme (ACE) that generates Ang II and inactivates the kinins. Inhibitors of ACE can reduce the impact of Ang II and potentiate the kinins, thus contributing to restore the cardiovascular homeostasis. In the last 20 years, ACE-inhibitors (ACE-Is) have become the drugs of first choice for the treatments of the major CVDs. ACE-Is not only reduce blood pressure, as sartans also do, but by protecting and potentiating the kinins, they can reduce morbidity and mortality and improve the quality of life for patients with CVDs. This paper provides a brief review of the literature on this topic.


Therapy, outcome and analysis of c-kit expression in patients with extrapulmonary small cell carcinoma.
In this study, we aimed to investigate the clinicopathological characteristics with special emphasis on c-kit expression and the treatment results of patients with extrapulmonary small cell carcinoma (EPSCC). The medical records of the patients with EPSCC were reviewed, and the data regarding patient and tumour characteristics, treatment and clinical outcome were retrieved and analysed. A total of 28 patients with the diagnosis of EPSCC were identified. There were 19 males and 9 females, with a mean age of 56.5 years. Patients with limited disease (LD) (n = 13) were treated with surgery, chemotherapy (CT) and radiotherapy with different sequences. Patients with extensive disease (ED) (n = 15) were mainly treated with combination CT. The median overall survival was 14.5 months in patients with LD compared to 11 months in those with ED (p = 0.029). Ten patients (36%) showed c-kit overexpression. There was no significant difference between the survival of c-kit-positive and c-kit-negative patients (p = 0.367). In conclusion, our study demonstrates that the prognosis of EPSCC is poor despite currently available treatments. C-kit may be considered as a potential target for novel therapeutical approaches.


MuSeeQ, a novel supervised image analysis tool for the simultaneous phenotyping of the soluble mucilage and seed morphometric parameters.
The mucilage is a model to study the polysaccharide biosynthesis since it is produced in large amounts and composed of complex polymers. In addition, it is of great economic interest for its technical and nutritional value. A fast method for phenotyping the released mucilage and the seed morphometric parameters will be useful for fundamental, food, pharmaceutical and breeding researches. Current strategies to phenotype soluble mucilage are restricted to visual evaluations or are highly time-consuming. Here, we developed a high-throughput phenotyping method for the simultaneous measurement of the soluble mucilage content released on a gel and the seed morphometric parameters. Within this context, we combined a biochemical assay and an open-source computer-aided image analysis tool, MuSeeQ. The biochemical assay consists in sowing seeds on an agarose medium containing the dye toluidine blue O, which specifically stains the mucilage once it is released on the gel. The second part of MuSeeQ is a macro developed in ImageJ allowing to quickly extract and analyse 11 morphometric data of seeds and their respective released mucilages. As an example, MuSeeQ was applied on a flax recombinant inbred lines population (previously screened for fatty acids content.) and revealed significant correlations between the soluble mucilage shape and the concentration of some fatty acids, e.g. C16:0 and C18:2. Other fatty acids were also found to correlate with the seed shape parameters, e.g. C18:0 and C18:2. MuSeeQ was then showed to be used for the analysis of other myxospermous species, including Arabidopsis thaliana and Camelina sativa. MuSeeQ is a low-cost and user-friendly method which may be used by breeders and researchers for phenotyping simultaneously seeds of specific cultivars, natural variants or mutants and their respective soluble mucilage area released on a gel. The script of MuSeeQ and video tutorials are freely available at http://MuSeeQ.free.fr.


The aging kidney: physiological changes.
Age-associated loss of kidney function has been recognized for decades. With aging, many subjects exhibit progressive decreases in glomerular filtration rate and renal blood flow, with wide variability among individuals. The fall in glomerular filtration rate is because of reductions in the glomerular capillary plasma flow rate and the glomerular capillary ultrafiltration coefficient. In addition, a primary reduction in afferent arteriolar resistance is associated with an increase in glomerular capillary hydraulic pressure. These hemodynamic changes occur in concert with structural changes, including loss of renal mass; hyalinization of afferent arterioles and in some cases, development of aglomerular arterioles; an increase in the percentage of sclerotic glomeruli; and tubulointerstitial fibrosis. Aging is associated with altered activity and responsiveness to vasoactive stimuli, such that responses to vasoconstrictor stimuli are enhanced, whereas vasodilatory responses are impaired. Changes in the activity of the renin-angiotensin and nitric oxide systems appear to be particularly important, as is the modulating effect of gender. These changes may predispose the older kidney to acute kidney injury, including normotensive ischemic nephropathy, as well as progressive chronic kidney disease.


Electrophysiological evidences of organization of cortical motor information in the Basal Ganglia.
During the last two decades, the many developments in the treatment of movement disorders such as Parkinson disease and dystonia have enhanced our understanding on organization of the basal ganglia, and this knowledge has led to other advances in the field. According to many electrophysiological and anatomical findings, it is considered that motor information from different cortical areas is processed through several cortico-basal ganglia loops principally in a parallel fashion and somatotopy from each cortical area is also well preserved in each loop. Moreover, recent studies suggest that not only the parallel processing but also some convergence of information occur through the basal ganglia. Information from cortical areas whose functions are close to each other tends to converge in the basal ganglia. The cortico-basal ganglia loops should be comprehended more as a network rather than as separated subdivisions. However, the functions of this convergence still remain unknown. It is important even for clinical doctors to be well informed about this kind of current knowledge because some symptoms of movement disorders may be explained by disorganization of the information network in the basal ganglia.


Vocal signature in wild infant chimpanzees.
A large array of communication signals supports the fission/fusion social organization in chimpanzees, and among them the acoustic channel plays a large part because of their forest habitat. Adult vocalizations convey social and ecological information to their recipients allowing them to obtain cues about an ongoing event from calls only. In contrast to adult vocalizations, information encoded in infant calls had been hardly investigated. Studies mainly focused on vocal development. The present article aims at assessing the acoustic cues that support individual identity coding in infant chimpanzees. By analyzing recordings performed in the wild from seven 3-year-old infant chimpanzees, we showed that their calls support a well-defined individual vocal signature relying on spectral cues. To assess the reliability of the signature across the calls of an individual, we defined two subsets of recordings on the basis of the characteristics of the frequency modulation (whimpers and screams) and showed that both call types present a reliable vocal signature. Early vocal signature may allow the mother and other individuals in the group to identify the infant caller when visual contact is broken. Chimpanzee mothers may have developed abilities to cope with changing vocal signatures while their infant, still vulnerable, gains in independence in close habitat.


Development of peripheral opioid antagonists' new insights into opioid effects.
The recent approval by the US Food and Drug Administration of 2 medications--methylnaltrexone and alvimopan--introduces a new class of therapeutic entities to clinicians. These peripherally acting mu-opioid receptor antagonists selectively reverse opioid actions mediated by receptors outside the central nervous system, while preserving centrally mediated analgesia. Methylnaltrexone, administered subcutaneously, has been approved in the United States, Europe, and Canada. In the United States, it is indicated for the treatment of opioid-induced constipation in patients with advanced illness (eg, cancer, AIDS) who are receiving palliative care, when response to laxative therapy has not been sufficient. Alvimopan, an orally administered medication, has been approved in the United States to facilitate recovery of gastrointestinal function after bowel resection and primary anastomosis. Clinical and laboratory studies performed during the development of these drugs have indicated that peripheral receptors mediate other opioid effects, including decreased gastric emptying, nausea and vomiting, pruritus, and urinary retention. Laboratory investigations with these compounds suggest that opioids affect fundamental cellular processes through mechanisms that were previously unknown. These mechanisms include modifications of human immunodeficiency virus penetration, tumor angiogenesis, vascular permeability, and bacterial virulence.


Reduction of plasma fibrinogen, immunoglobulin G, and immunoglobulin M concentrations by immunoadsorption therapy with tryptophan and phenylalanine adsorbents.
Immunoadsorption (IA) therapy with tryptophan (TR-350) or phenylalanine (PH-350) adsorbents has been used to reduce the concentration of serum antibodies in human lymphocyte antigen (HLA)-immunized patients. Other forms of plasma purification have been reported to reduce the level of fibrinogen, which affects the blood properties. In this study we investigated the effects of IA therapy using both adsorbents on plasma fibrinogen and immunoglobulins G and M in 13 patients (8 patients were treated with TR-350, and 5 patients were treated with PH-350). During each session I plasma volume (2.8 +/- 0.4 L of plasma) was processed through the immunocolumn and then returned to the patient together with the blood cells. Compared with the pretreatment values, the plasma fibrinogen, IgG, and IgM concentrations were significantly reduced after IA therapy (p < 0.01 for TR-350; p < 0.04 for PH-350). There was a positive correlation between the degree of reduction of plasma proteins and the number of IA treatments given. A nonparametric test (Wilcoxon's signed-rank test or the Mann-Whitney test) was used for statistical analysis. We conclude from our study that IA therapy effectively lowers the plasma levels of fibrinogen, IgG, and IgM and thus can be considered a valuable alternative to other blood purification methods.


Transcriptional and posttranscriptional regulation of transcription factor expression in Arabidopsis roots.
Understanding how the expression of transcription factor (TF) genes is modulated is essential for reconstructing gene regulatory networks. There is increasing evidence that sequences other than upstream noncoding can contribute to modulating gene expression, but how frequently they do so remains unclear. Here, we investigated the regulation of TFs expressed in a tissue-enriched manner in Arabidopsis roots. For 61 TFs, we created GFP reporter constructs driven by each TF's upstream noncoding sequence (including the 5'UTR) fused to the GFP reporter gene alone or together with the TF's coding sequence. We compared the visually detectable GFP patterns with endogenous mRNA expression patterns, as defined by a genome-wide microarray root expression map. An automated image analysis method for quantifying GFP signals in different tissues was developed and used to validate our visual comparison method. From these combined analyses, we found that (i) the upstream noncoding sequence was sufficient to recapitulate the mRNA expression pattern for 80% (35/44) of the TFs, and (ii) 25% of the TFs undergo posttranscriptional regulation via microRNA-mediated mRNA degradation (2/24) or via intercellular protein movement (6/24). The results suggest that, for Arabidopsis TFs, upstream noncoding sequences are major contributors to mRNA expression pattern establishment, but modulation of transcription factor protein expression pattern after transcription is relatively frequent. This study provides a systematic overview of regulation of TF expression at a cellular level.


PKCθ regulates T cell motility via ezrin-radixin-moesin localization to the uropod.
Cell motility is a fundamental process crucial for function in many cell types, including T cells. T cell motility is critical for T cell-mediated immune responses, including initiation, activation, and effector function. While many extracellular receptors and cytoskeletal regulators have been shown to control T cell migration, relatively few signaling mediators have been identified that can modulate T cell motility. In this study, we find a previously unknown role for PKCθ in regulating T cell migration to lymph nodes. PKCθ localizes to the migrating T cell uropod and regulates localization of the MTOC, CD43 and ERM proteins to the uropod. Furthermore, PKCθ-deficient T cells are less responsive to chemokine induced migration and are defective in migration to lymph nodes. Our results reveal a novel role for PKCθ in regulating T cell migration and demonstrate that PKCθ signals downstream of CCR7 to regulate protein localization and uropod formation.


The retaining system of the face: histologic evaluation of the septal boundaries of the subcutaneous fat compartments.
Because the concept of subcutaneous fat compartments has many significant implications for cosmetic and reconstructive surgery, it is important to verify the original findings and validate the concept. The authors studied the histology of the septal boundaries between several adjacent fat compartments. Eighteen hemifacial cadaver specimens were used (five male and four female cadavers; age range, 39 to 87 years). Tissue marking dye was injected into the central forehead and the medial, middle, and lateral temporal cheek compartments. Dye was allowed to diffuse for 4 hours until a skin blush was noted, at which point dye-setting solution was injected to fix the dye. En bloc transverse specimens were harvested and stored in formalin overnight. Standard histologic processing was performed. Each compartment partitioned dye in a consistent and reproducible manner. A fibrous condensation of connective tissue formed the diffusion barriers. These septa originated from underlying fascia and inserted into the dermis of the skin. A septal barrier originated from the fascia of the frontalis muscle, so these septal barriers are not necessarily related to the superficial musculoaponeurotic system but can occur anywhere between superficial fascia and skin. These findings support the concept that subcutaneous fat is compartmentalized, specifically by fascial condensations that travel from superficial fascia to dermis. These septa form an interconnecting framework that limits shearing forces on the face. This framework provides a "retaining system" for the human face. Implicit in this concept is the suggestion that the face ages three dimensionally, with separate compartments changing relative to one another by both position and volume.


Ventralized zebrafish embryo rescue by overexpression of Zic2a.
The neuroectoderm arises during gastrulation as a population of undifferentiated proliferating neuroepithelial cells. As development continues, neuroepithelial cells leave the cell cycle and differentiate into neurons and glia of the functioning central nervous system. What processes establish the spatial distribution of proliferating neuroepithelial cells? To investigate this question, zic2a was isolated from zebrafish, a homolog of the Drosophila pair-rule gene odd-paired, which is involved in nervous system patterning. At shield stage, zic2a was expressed in the zebrafish organizer and the blastoderm margin, and became restricted to the axial mesoderm in mid-gastrula. Expression of zic2a appeared in the prospective neuroectoderm during gastrulation, and later demarcated the presumptive forebrain. This expression pattern suggests that zic2a may function early in the organizer and later in the neural plate to demarcate the population of proliferating neuroectoderm. Consistent with a function for zic2a in transducing signals from the organizer, overexpression of zic2a resulted in an expansion of proliferating neuroectoderm. Furthermore, zic2a overexpression rescued the ventralized phenotype of chordino mutant embryos, which lack a functional chordin gene. Early expression of zic2 in the zebrafish organizer, and the phenotype resulting from overexpression, show a role for zic2a downstream of chordin or other secreted organizer proteins in establishing the initial size of the population of neuroectoderm cells.


Neonatal myotubular myopathy with respiratory distress: report of a case.
The case of a 2-day-old male full-term newborn with myotubular (centronuclear) myopathy is reported. He presented with generalized hypotonia and muscle weakness, swallowing disturbance, and respiratory distress at birth. He had a typical myopathic face, high-arched palate, funnel chest, and mild bilateral ptosis. Deep tendon reflexes were absent. Serum creatine kinase was normal. The histologic examination of the muscle biopsy over the right rectus femoris muscle revealed an increased number of fibers with centrally placed nuclei, type 1 fiber predominance, type 1 fiber atrophy, and a peripheral halo in the sarcoplasm on NADH-TR staining. On electron microscopy, central nuclei were separated by strands of glycogen and mitochondria. His muscle strength showed clinical improvement at a 14-month follow-up. This case illustrates the need for a muscle biopsy and histochemical staining and/or electron microscopic investigation for a proper diagnosis in hypotonic newborns with respiratory distress.


Influence of cardiorespiratory fitness on the association between C-reactive protein and metabolic syndrome prevalence in racially diverse women.
Metabolic syndrome and C-reactive protein (CRP) are independent predictors of cardiovascular disease (CVD) among women. The extent to which cardiorespiratory fitness influences the relationship between CRP and metabolic syndrome is unknown. Cross-sectional associations among fitness, CRP, and metabolic syndrome were examined in 135 African American, Native American, and Caucasian women (55 +/- 11 years, 28 +/- 6 kg/m2). Fitness was quantified with a symptom-limited maximal treadmill exercise test. Plasma CRP concentrations were determined with the Dade-Behring high-sensitivity immunoassay. Metabolic syndrome was defined according to NCEP-ATP III. Metabolic syndrome, CRP, and fitness varied (p < 0.05) by race. Race-adjusted CRP values were directly associated (p < 0.05) with each metabolic syndrome component. After adjusting for age and race, the relative odds of metabolic syndrome was 3.6 (95% CI = 1.5 - 8.4) in women with elevated (> 2.0 mg/L) vs. low CRP. Adjustment for smoking, hormone therapy, body mass index (BMI), and HOMA insulin resistance did not eliminate this association (p < 0.05). The association between CRP and the metabolic syndrome was no longer significant (OR = 1.3, 95% CI = 0.9 - 5.9, p = 0.59) after adjustment for fitness. Higher cardiorespiratory fitness may be an important consideration in the milieu of vascular inflammation and metabolic syndrome.


Inhaled Amikacin to Prevent Ventilator-Associated Pneumonia.
Whether preventive inhaled antibiotics may reduce the incidence of ventilator-associated pneumonia is unclear. In this investigator-initiated, multicenter, double-blind, randomized, controlled, superiority trial, we assigned critically ill adults who had been undergoing invasive mechanical ventilation for at least 72 hours to receive inhaled amikacin at a dose of 20 mg per kilogram of ideal body weight once daily or to receive placebo for 3 days. The primary outcome was a first episode of ventilator-associated pneumonia during 28 days of follow-up. Safety was assessed. A total of 850 patients underwent randomization, and 847 were included in the analyses (417 assigned to the amikacin group and 430 to the placebo group). All three daily nebulizations were received by 337 patients (81%) in the amikacin group and 355 patients (83%) in the placebo group. At 28 days, ventilator-associated pneumonia had developed in 62 patients (15%) in the amikacin group and in 95 patients (22%) in the placebo group (difference in restricted mean survival time to ventilator-associated pneumonia, 1.5 days; 95% confidence interval [CI], 0.6 to 2.5; P = 0.004). An infection-related ventilator-associated complication occurred in 74 patients (18%) in the amikacin group and in 111 patients (26%) in the placebo group (hazard ratio, 0.66; 95% CI, 0.50 to 0.89). Trial-related serious adverse effects were seen in 7 patients (1.7%) in the amikacin group and in 4 patients (0.9%) in the placebo group. Among patients who had undergone mechanical ventilation for at least 3 days, a subsequent 3-day course of inhaled amikacin reduced the burden of ventilator-associated pneumonia during 28 days of follow-up. (Funded by the French Ministry of Health; AMIKINHAL ClinicalTrials.gov number, NCT03149640; EUDRA Clinical Trials number, 2016-001054-17.).


Characterisation of exposure to non-ionising electromagnetic fields in the Spanish INMA birth cohort: study protocol.
Analysis of the association between exposure to electromagnetic fields of non-ionising radiation (EMF-NIR) and health in children and adolescents is hindered by the limited availability of data, mainly due to the difficulties on the exposure assessment. This study protocol describes the methodologies used for characterising exposure of children to EMF-NIR in the INMA (INfancia y Medio Ambiente- Environment and Childhood) Project, a prospective cohort study. Indirect (proximity to emission sources, questionnaires on sources use and geospatial propagation models) and direct methods (spot and fixed longer-term measurements and personal measurements) were conducted in order to assess exposure levels of study participants aged between 7 and 18 years old. The methodology used varies depending on the frequency of the EMF-NIR and the environment (homes, schools and parks). Questionnaires assessed the use of sources contributing both to Extremely Low Frequency (ELF) and Radiofrequency (RF) exposure levels. Geospatial propagation models (NISMap) are implemented and validated for environmental outdoor sources of RFs using spot measurements. Spot and fixed longer-term ELF and RF measurements were done in the environments where children spend most of the time. Moreover, personal measurements were taken in order to assess individual exposure to RF. The exposure data are used to explore their relationships with proximity and/or use of EMF-NIR sources. Characterisation of the EMF-NIR exposure by this combination of methods is intended to overcome problems encountered in other research. The assessment of exposure of INMA cohort children and adolescents living in different regions of Spain to the full frequency range of EMF-NIR extends the characterisation of environmental exposures in this cohort. Together with other data obtained in the project, on socioeconomic and family characteristics and development of the children and adolescents, this will enable to evaluate the complex interaction between health outcomes in children and adolescents and the various environmental factors that surround them.


Oral warfarin affects peripheral blood leukocyte IL-6 and TNFα production in rats.
Warfarin is a Vitamin K (VK) antagonist that affects Vitamin K-dependent (VKD) processes, including blood coagulation, as well as processes unrelated to hemostasis such as bone growth, calcification, and growth of some cell types. In addition, warfarin exerts influence on some non-VKD-related activities, including anti-tumor and immunomodulating activity. With respect to the latter, both immune stimulating and suppressive effects have been noted in different experimental systems. To explore the in vivo immunomodulatory potential of warfarin on one type of activity (i.e., cytokine production) in two different immune cell populations (i.e., mononuclear or polymorphonuclear cells), effects of subchronic oral warfarin intake in rats on pro-inflammatory cytokine (i.e., TNFα, IL-6) production by peripheral blood mononuclear and polymorphonuclear cells (granulocytes) was examined. Differential effects of warfarin intake on TNFα and IL-6 were noted, depending on the type of peripheral blood leukocytes and on the cytokine examined. Specifically, a lack of effect on TNFα and a priming of IL-6 production by mononuclear cells along with a decrease in TNFα and a lack of effect on IL-6 in polymorphonuclear cells were seen in warfarin-exposed hosts. The cell- and cytokine-dependent effects from subchronic oral warfarin intake on peripheral blood leukocytes demonstrated in this study could, possibly, differentially affect reactions mediated by these cells. Ultimately, the observed effects in rats might have implications for those humans who are on long-term/prolonged warfarin therapy.


Immunoglobulin specificity of TG19318: a novel synthetic ligand for antibody affinity purification.
A synthetic ligand [TG19318], able to mimic protein A in the recognition of the immunoglobulin Fc portion, has been previously identified in our laboratory through the synthesis and screening of multimeric combinatorial peptide libraries. In this study we have fully characterized its applicability in affinity chromatography for the downstream processing of antibodies, examining the specificity and selectivity for polyclonal and monoclonal immunoglobulins derived from different sources. Ligand specificity was broader than protein A, since IgG deriving from human, cow, horse, pig, mouse, rat, rabbit, goat and sheep sera, IgY obtained from egg yolk, and IgM, IgA and IgE were efficiently purified on TG19318 affinity columns. Adsorbed antibodies were conveniently eluted by a buffer change to 0.1 M acetic acid or 0.1 M sodium bicarbonate pH 9, with full retention of immunological properties. Monoclonal antibodies deriving from cell culture supernatants or ascitic fluids were also conveniently purified on TG19318 affinity columns, even from very diluted samples. The affinity constant for the TG19318-IgG interaction was 0.3 microM, as determined by optical biosensor measurements. Under optimized conditions, antibody purity after affinity purification was close to 95%, as determined by densitometric scanning of SDS-PAGE gels of purified fractions, and maximal column capacity reached 25 mg Ig/ml support. In vivo toxicity studies in mice indicated a ligand oral toxicity greater than 2000 mg kg-1 while intravenous toxicity was close to 150 mg kg-1. Validation of antibody affinity purification processes for therapeutic use, a very complex, laborious and costly procedure, is going to be simplified by the use of TG19318, which could reduce considerably the presence of biological contaminants in the purified preparation, a very recurrent problem when using recombinant or extractive biomolecules as affinity ligands.


Cell cycle implication on nitrogen acquisition and synchronization in Thalassiosira weissflogii (Bacillariophyceae).
The Michaelis-Menten model of nitrogen (N) acquisition, originally used to represent the effect of nutrient concentration on the phytoplankton uptake rate, is inadequate when other factors show temporal variations. Literature generally links diurnal oscillations of N acquisition to a response of the physiological status of microalgae to photon flux density (PFD) and substrate availability. This work describes how the cell cycle also constitutes a significant determinant of N acquisition and, when appropriate, assesses the impact of this property at the macroscopic level. For this purpose, we carried out continuous culture experiments with the diatom Thalassiosira weissflogii (Grunow) G. Fryxell & Hasle exposed to various conditions of light and N supply. The results revealed that a decrease in N acquisition occurred when a significant proportion of the population was in mitosis. This observation suggests that N acquisition is incompatible with mitosis and therefore that its acquisition rate is not constant during the cell cycle. In addition, environmental conditions, such as light and nutrient supply disrupt the cell cycle at the level of the individual cell, which impacts synchrony of the population.


Identification, cloning, and expression of a cytosolic megakaryocyte protein-tyrosine-phosphatase with sequence homology to cytoskeletal protein 4.1.
We have isolated a cDNA encoding a third type of protein-tyrosine-phosphatase. We screened human megakaryoblastic cell line (MEG-01) an umbilical vein endothelial cell cDNA libraries to obtain a 3.7-kilobase cDNA designated PTPase MEG. Northern blot analysis of MEG-01 RNA detected a 3.7-kilobase transcript, suggesting that a full-length cDNA has been identified. PTPase MEG cDNA contains an open reading frame of 926 amino acids. The cDNA has a G+C-rich 5' untranslated region of 771 nucleotides that has the potential to form stable stem-loop structures and has two upstream ATG codons. The predicted protein (Mr = 105,910) has no apparent membrane-spanning region and contains a single protein-tyrosine-phosphatase domain (amino acids 659-909) that is 35-40% identical to previously described tyrosine-phosphatase domains. The recombinant phosphatase domain possesses protein-tyrosine-phosphatase activity when expressed in Escherichia coli. The amino-terminal region (amino acids 31-367) is 45% identical to the amino terminus of human erythrocyte protein 4.1, a cytoskeletal protein. The identification of a protein-tyrosine-phosphatase that is related to cytoskeletal proteins implies that cell signaling activities reside not only in transmembrane receptors but in cytoskeletal elements as well.


Phosphorylation of 4.1N by CaMKII Regulates the Trafficking of GluA1-containing AMPA Receptors During Long-term Potentiation in Acute Rat Hippocampal Brain Slices.
GluA1-containing α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors (AMPARs) inserted into postsynaptic membranes are key to the process of long-term potentiation (LTP). Some evidence has shown that 4.1N plays a critical role in the membrane trafficking of AMPARs. However, the underlying mechanism behind this is still unclear. We investigated the role of 4.1N-mediated membrane trafficking of AMPARs during theta-burst stimulation long-term potentiation (TBS-LTP), to illustrate the molecular mechanism behind LTP. LTP was induced by TBS in rat hippocampal CA1 neuron. Tat-GluA1 (MPR), which disrupts the association of 4.1N-GluA1, and autocamtide-2-inhibitory peptide, myristoylated (Myr-AIP), a CaMKII antagonist, were used to explore the role of 4.1N in the AMPARs trafficking during TBS-induced LTP. Immunoprecipitation (IP) and immunoblotting (IB)were used to detect protein expression, phosphorylation, and the interaction of p-CaMKII-4.1N-GluA1. We found that Myr-AIP attenuated increases of p-CaMKII (T286), p-GluA1 (ser831), and 4.1N phosphorylation after TBS-LTP, and decreased the association of p-CaMKII-4.1N-GluA1, along with the expression of GluA1, at postsynaptic densities during TBS-LTP. We also designed interfering peptides to disrupt the interaction between 4.1N and GluA1, which showed that Tat-GluA1 (MPR) or Myr-AIP inhibited TBS-LTP and attenuated increases of GluA1 at postsynaptic sites, while Tat-GluA1 (MPR) or Myr-AIP had no effects on miniature excitatory postsynaptic currents (mEPSCs) in non-stimulated hippocampal CA1 neurons. Active CaMKII enhanced the phosphorylation of 4.1N and facilitated the association of p-CaMKII with 4.1N-GluA1, which in turn resulted in GluA1 trafficking during TBS-LTP. The association of 4.1N-GluA1 is required for LTP, but not for basal synaptic transmission.


Enhanced antigenicity of autologous leukemia cells enriched with cholesterylhemisuccinate.
Cholesterylhemisuccinate (CHS) has been used by different authors to enhance the antigenicity of tumor cells in human and in animals. In the present study, leukemic cells isolated mainly from patients with chronic myeloid and chronic lymphocytic leukemia were incubated for 90 min at 37 degrees in PBS containing 3.5% polyvinylpyrrolidone, 0.5% glucose, 1% human serum albumin and 150 micrograms/ml CHS 5 x 10(6) pretreated, irradiated, autologous leukemic cells were tested for their ability to elicit a delayed type hypersensitivity skin reaction. The negative controls included CHS-treated and untreated non-leukemic autologous blood cells as well as substance controls. Recall antigens such as tetanus, diphtheria and tuberculine included in the Multitest Mérieux served as positive controls. Results from 28 patients and 44 skin tests are presented. 19 of 23 patients tested under the best of experimental conditions generated a positive skin reaction with the CHS-treated autologous leukemic cells. Less than 12% of the patients reacted with any of the above negative controls. Under the same conditions, more than 90% of the CLL patients were reactive. 5 CLL patients were selected for a treatment including 6 weekly injections of 5 x 10(6) to 5 x 10(8) autologous CLL cells. 3 patients experienced an up to 60% reduction of their initial leukocyte counts. In one patient, the leukocyte counts continued to decrease more than 6 months after the treatment was completed. One patient showed no response and in one patient with increasing leukocytes this increase could not be reversed. The results are discussed in terms of earlier observations that membrane sterol content influences the presentation of cell surface proteins.


Ruvbl1 is Essential for Ciliary Beating during Xenopus laevis Embryogenesis.
The Ruvb-like AAA ATPase1 (Ruvbl1; also known as Pontin) is an evolutionary conserved protein belonging to the adenosine triphosphates associated with diverse cellular activities (AAA+) superfamily of ATPases. Ruvbl1 is a component of various protein supercomplexes and is involved in a variety of cellular activities, including chromatin remodeling, DNA damage repair, and mitotic spindle assembly however, the developmental significance of this protein is unknown and needs detailed investigation. We investigated the developmental significance of Ruvbl1 in multiciliated cells of the Xenopus laevis epidermis since ruvbl1 is expressed in the multiciliated cells and pronephros during X. laevis embryogenesis. The knockdown of ruvbl1 significantly impaired cilia-driven fluid flow and basal body polarity in the X. laevis epidermis compared to control embryos, but did not affect cilia morphology. Our results suggest that Ruvbl1 plays a significant role in embryonic development by regulating ciliary beating; however, further investigation is needed to determine the mechanisms involved.


Neuraxial anesthesia is associated with improved survival after total joint arthroplasty depending on frailty: a cohort study.
Frailty increases risk for complications after total joint arthroplasty (TJA). Whether this association is influenced by anesthetic administered is unknown. We hypothesized that use of neuraxial (spinal or epidural) anesthesia is associated with better outcomes compared with general anesthesia, and that the effect of anesthesia type on outcomes differs by frailty status. This single-institution cohort study included all patients (≥50 years) from January 2005 through December 2016 undergoing unilateral, primary and revision TJA. Using multivariable Cox regression, we assessed relationships between anesthesia type, a preoperative frailty deficit index (FI) categorized as non-frail (FI <0.11), vulnerable (FI 0.11 to 0.20), and frail (FI >0.20), and complications (mortality, infection, wound complications/hematoma, reoperation, dislocation, and periprosthetic fracture) within 1 year after surgery. Interactions between anesthesia type and frailty were tested, and stratified models were presented when an interaction (p<0.1) was observed. Among 18 458 patients undergoing TJA, more patients were classified as frail (21.5%) and vulnerable (36.2%) than non-frail (42.3%). Anesthesia type was not associated with complications after adjusting for age, joint, and revision type. However, in analyzes stratified by frailty, vulnerable patients under neuraxial block had less mortality (HR=0.49; 95% CI 0.27 to 0.89) and wound complications/hematoma (HR=0.71; 95% CI 0.55 to 0.90), whereas no difference in risk by anesthesia type was observed among patients found non-frail or frail. Neuraxial anesthesia use among vulnerable patients was associated with improved survival and less wound complications. Calculating preoperative frailty prior to TJA informs perioperative risk and enhances shared-decision making for selection of anesthesia type.


Absolute phase birefringence dispersion in polarization-maintaining fiber or birefringent crystal retrieved from a channeled spectrum.
We report on a simple method for retrieving the wavelength dependence of the phase birefringence in a polarization-maintaining fiber or a birefringent crystal from a channeled spectrum. The method utilizes interference of polarized modes or waves resolved as the channeled spectrum and its processing by a windowed Fourier transform to reconstruct precisely the phase as a function of wavelength. The ambiguity of the phase is removed provided that we know both the approximative function for the birefringence dispersion and the length of the fiber or the thickness of the crystal. The method is used in measuring the wavelength dependence of the phase birefringence in an elliptical-core fiber or in a quartz crystal in a range from 500 to 900 nm. The dependences are compared with those resulting from the available data, and very good agreement is confirmed.


Geriatrics grand rounds: Eve's rib, or a revisionist view of osteoporosis in men.
It is widely accepted that estrogen withdrawal following menopause predisposes women to accelerated bone loss and increased risk of developing osteoporosis. Although osteoporosis is a significant public health problem for aging men as well as women, the cause of osteoporosis in men remains largely unknown. A substantial number of men with osteoporosis present with bone loss secondary to conditions associated with reduced gonadal steroid hormone levels. Although hypogonadism is related to bone loss in men, and androgen levels decline with age in men, it is not at all clear that reduced androgen levels are related to bone loss in older men. What, then, is the role of gonadal steroids in osteoporosis in men? This review focuses on recent research--including clinical investigations of men with genetic disorders of estrogen action, basic biomedical studies of estrogen receptor "knockout" mice, and population-based comparisons of bone density with gonadal steroids in older men--leading to the surprising conclusion that estrogen plays a vital role in maintenance of bone in men as well as in women. Possible mechanisms whereby reduced estrogen levels might result in bone loss in both sexes are also reviewed, as are potential therapeutic implications of a role for estrogen in osteoporosis in men.


Quetiapine as an alternative to clozapine in the treatment of dopamimetic psychosis in patients with Parkinson's disease.
There are many difficulties associated with the late stages of Parkinson's disease (PD), but psychosis and agitation may be the most disturbing for both patients and care givers, and often precipitate the pivotal decision for long-term nursing home placement. While the addition of antipsychotic drugs or the withdrawal of antiparkinsonian drugs may improve the behavioral problem, these strategies usually worsen the motor difficulties. Clozapine has been studied in PD for over a decade, and while it appears to be effective, there are safety and tolerability concerns associated with it. In addition, in New Jersey, Medicaid no longer pays for the home blood draws that are required for home-bound patients. This led to a situation in which we had patients who needed to stop clozapine and begin an alternative therapy. Because quetiapine seems particularly well suited to patients with PD based on in vitro and in vivo studies we have begun to try this medication in PD patients who need to stop clozapine. This article reports three case histories of patients with PD, confusion and dopamimetic psychosis who had been previously managed with clozapine and who were successfully switched to quetiapine. At doses from 12.5 to 150 mg/day quetiapine was well tolerated, resulting in behavioral improvement and no real increase in parkinsonism. These case histories raise the possibility that quetiapine may represent a viable alternative to clozapine in PD patients with dopamimetic psychosis and behavioral disturbances.


Pregabalin treatment for neuropathic pain may damage intervertebral disc tissue.
The aim of the present study was to determine whether pharmaceutical preparations with pregabalin (PGB) as an active ingredient, which are widely prescribed by clinicians, exert toxic effects on human primary nucleus pulposus (NP) and annulus fibrosis (AF). Primary human cell cultures were obtained from intact (n=6) and degenerated (n=6) tissues resected from the two groups of patients. Different doses of PGB were applied to these cultures and cells were subjected to molecular analyses at 0, 24 and 48 h. Cell vitality, toxicity and proliferation were assessed using a spectrophotometer. The expression of chondroadherin (CHAD), a (member of the NP-specific protein family), hypoxia-inducible factor-1α (HIF-1α) and type II collagen (COL2A1) was measured using reverse transcription-quantitative polymerase chain reaction. The results revealed that cell intensity increased in a time-dependent manner and cell vitality continued in the cultures without pharmaceuticals. Cell proliferation was suppressed in the PGB-treated cultures independent from the dose and duration of application. PGB was demonstrated to suppress the expression of CHAD and HIF-1α. In contrast, COL2A1 gene expression was not revealed in any experimental group. The present study utilized an in vitro model and the PGB active ingredient used herein may not be representative of clinical applications; however, the results demonstrated that PGB has a toxic effect on NP/AF cell cultures containing primary human intervertebral disc tissue. In summary, the use of pharmacological agents containing PGB may suppress the proliferation and differentiation of NP/AF cells and/or tissues, which should be considered when deciding on an appropriate treatment regime.


Isolation of nuclear acidic proteins from rat tissues. Characterization of acetylated liver nuclear acidic proteins.
Nuclear acidic proteins isolated from rat brain, heart, kidney and liver showed similar, complex patterns on electrophoresis in sodium dodecyl sulphate-polyacrylamide gels. The contamination of nuclear acidic proteins by nuclear-membrane acidic proteins was found to the extent of 11%. Incorporation of [(3)H]acetate into the various nuclear acidic proteins in vivo, which were fractionated by polyacrylamide-gel electrophoresis, differed from tissue to tissue. Hydrolysis of these acetylated nuclear acidic proteins with 6m-HCl at 110 degrees C released 70% of the radioactivity, which indicated that labile acetyl groups had been incorporated into these proteins. Analysis of [(3)H]acetate-labelled nuclear acidic proteins revealed two acetylated amino acid residues, N(2)-acetylserine and N(2)-acetyl-lysine. The significance of the role played by nuclear acidic proteins in relation to gene regulation is discussed.


[Effect of nuciferine on gut microbiota and inflammatory response in obese model mice].
The aim of this study was to elucidate the mechanism of nuciferine on alleviating obesity based on modulating gut microbiota, ameliorating chronic inflammation, and improving gut permeability. In this study, the obese model mice were induced by high-fat diet and then randomly divided into model group, and nuciferine group; some other mice of the same week age were fed with normal diet as normal group. In the modeling process, the mice were administered intragastrically(ig) for 12 weeks. In the course of both modeling and treatment, the body weight and food intake of mice in each group were measured weekly. After modeling and treatment, the Lee's index, weight percentage of inguinal subcutaneous fat, and the level of blood lipid in each group were measured. The pathological changes of adipocytes were observed by HE staining to evaluate the efficacy of nuciferine treatment in obese model mice. 16 S rRNA sequencing analysis was conducted to study the changes in diversity and abundance of gut microbiota after nuciferine treatment. Enzyme-linked immunosorbent assay(ELISA) and quantitative Real-time polymerase chain reaction(qPCR) were used to detect the levels of inflammatory factors interleukin-6(IL-6), interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α) and the expression of related genes in adipose tissue of mice in each group, so as to evaluate the effect of nuciferine on chronic inflammation of mice in obese model group. qPCR was used to detect the expression of occludin and tight junction protein 1(ZO-1)gene in colon tissure, so as to evaluate the effect of nuciferine on intestinal permeability of mice in obese group. Nuciferine decreased the body weight of obese mice, Lee's index, weight percentage of inguinal subcutaneous fat(P<0.05), and reduced the volume of adipocytes, decreased the level of total cholesterol(TC), triglyceride(TG), and low density lipoprotein cholesterol(LDL-C)(P<0.05) in serum, improved dysbacteriosis, increased the relative abundance of Alloprevotella, Turicibacter, and Lactobacillus, lowered the relative abundance of Helicobac-ter, decreased the expression of inflammatory cytokines IL-6, IL-1β, and TNF-α genes in adipose tissue(P<0.01), decreased the levels of inflammatory cytokines IL-6, IL-1β, and TNF-α in serum(P<0.05), and increased the expression of occludin and ZO-1 genes related to tight junction in colon tissue(P<0.01). Nuciferine could treat obesity through modulating gut microbiota, decreasing gut permeability and ameliorating inflammation.


Can zirconia crowns be the first restorative choice after endodontic treatment of primary teeth?
Early loss of primary teeth due to caries can cause temporary or permanent problems, such as occlusion disorders, aesthetic problems, nutritional problems, abnormal habits, and speech disorders. Although the development of restorative materials has led to significant improvements, failures are still frequent. The purpose of this study was to evaluate the success rates of zirconia crowns compared to other crowns for the restoration of endodontically treated teeth with excessive loss of material. A total of 45 teeth of patients aged under 10 years, including primary molars and anterior teeth, requiring restoration after root canal treatment, were included in this randomized, non-blinded clinical study. Stainless steel (SSC), zirconia (ZC), and strip (SC) crowns were used and compared clinically and radiographically during a six-month follow-up period. Parents were asked to fill out a questionnaire at one week and six months to evaluate the parent and child satisfaction levels. The patients' pain and discomfort levels were assessed using a visual analogue scale. The Mann-Whitney U, Chi-Squared and Friedman and Cochran Q tests were used for statistical comparisons using IBM SPSS Statistics version 23 (IBM, Armonk, NY, USA). The six-month follow-up evaluations revealed no significant differences in clinical outcomes between the SSC, ZC, and SC groups. There were no significant differences between the groups with respect to parent and child satisfaction. The clinical outcomes of ZCs were comparable to those of the other crowns. ZCs could be a choice for aesthetic purposes, with the disadvantage of a higher cost.


[Intellectual occupations and chronic stress in shift work].
Genesis of chronic occupational stress covers steps of sequent transformation from functional state to strain, fatigue and overfatigue. Physiologic features of these conditions were analyzed, so the conditions could be assessed through functional levels, relationships between central nervous and cardiovascular systems and inner links of these systems, coordination of diurnal rhythms. Workers having higher occupational psychoemotional strain, shift work schedule and 12-hours working day are significantly prone to neurotic disorders.


A CASP-8 mutation recognized by cytolytic T lymphocytes on a human head and neck carcinoma.
Of the antigens recognized on human tumors by autologous cytolytic T lymphocytes, all those defined thus far have been identified on melanoma or renal cell carcinoma. We report here the identification of an antigen recognized by autologous cytolytic T lymphocytes on a human squamous cell carcinoma of the oral cavity. The antigen is encoded by a mutated form of the CASP-8 gene. This gene, also named FLICE or MACH, codes for protease caspase-8, which is required for induction of apoptosis through the Fas receptor and tumor necrosis factor receptor-1. The mutation, which was found in the tumor cells but not in the normal cells of the patient, modifies the stop codon and adds an Alu repeat to the coding region, thereby lengthening the protein by 88 amino acids. The ability of the altered protein to trigger apoptosis appears to be reduced relative to the normal caspase-8. The antigenic peptide is a nonamer presented by HLA-B*3503. The five last amino acids are encoded by the extension of the reading frame caused by the mutation. This, together with previous observations of CDK4 and beta-catenin mutations, suggests that a significant fraction of the point mutations generating a tumor antigen also play a role in the tumoral transformation or progression.


Effects of Virtual Reality Therapy on Gait and Balance Among Individuals With Spinal Cord Injury: A Systematic Review and Meta-analysis.
Background and Purpose. The use of virtual reality (VR) therapy among individuals with spinal cord injury (SCI) is a relatively new rehabilitation approach replicating real-life scenarios. The aim of this study was to evaluate the effectiveness of VR therapy for improving gait and balance in individuals with SCI. Methods. Databases of PubMed, Web of Science, Scopus, SportDiscuss, and CINHAL were searched from inception until September 2019. Two independent reviewers screened articles for inclusion, extracted data, and evaluated methodological quality of the trials. Results. Ten trials, including 3 randomized clinical trials (RCTs) and 7 pre-post trials, with a total of 149 participants were analyzed. Meta-analysis of RCTs demonstrated significant improvement in sitting balance (standardized mean difference [SMD] = 1.65; 95% CI 1.21-2.09; P < .01) after VR therapy with conventional rehabilitation compared with only conventional rehabilitation. Also, pre-post studies using VR therapy without a control group showed improvement in standing balance (Berg Balance Scale, MD = 4.22; 95% CI 1.78-6.66; P < .01 and Activities-specific Balance Confidence scale, MD = 8.53; 95% CI 2.52-14.53; P = .01) and a trend for improvement in gait (SMD = 0.34; 95% CI 0.02-0.66; P = .04). Conclusion. This study demonstrated the beneficial effects of VR therapy to enhance sitting and standing balance and showed a trend of gait improvement in individuals with SCI. This conclusion is based on mainly preliminary data and therefore, more RCTs are needed to confirm the effects of the use of VR in individuals with SCI.


Antiviral response dictated by choreographed cascade of transcription factors.
The dendritic cell (DC) is a master regulator of immune responses. Pathogenic viruses subvert normal immune function in DCs through the expression of immune antagonists. Understanding how these antagonists interact with the host immune system requires knowledge of the underlying genetic regulatory network that operates during an uninhibited antiviral response. To isolate and identify this network, we studied DCs infected with Newcastle disease virus, which is able to stimulate innate immunity and DC maturation through activation of RIG-I signaling, but lacks the ability to evade the human IFN response. To analyze this experimental model, we developed a new approach integrating genome-wide expression kinetics and time-dependent promoter analysis. We found that the genetic program underlying the antiviral cell-state transition during the first 18 h postinfection could be explained by a single convergent regulatory network. Gene expression changes were driven by a stepwise multifactor cascading control mechanism, where the specific transcription factors controlling expression changed over time. Within this network, most individual genes were regulated by multiple factors, indicating robustness against virus-encoded immune evasion genes. In addition to effectively recapitulating current biological knowledge, we predicted, and validated experimentally, antiviral roles for several novel transcription factors. More generally, our results show how a genetic program can be temporally controlled through a single regulatory network to achieve the large-scale genetic reprogramming characteristic of cell-state transitions.


Hypomania related to phenelzine and isoetharine interaction in one patient.
A case is reported of a man with apparent unipolar depression that was responsive to treatment with phenelzine who became hypomanic when isoetharine was added to treat his chronic obstructive pulmonary disease. The role of beta-adrenergic receptors in affective illness is reviewed in light of this case.


The impact of national health insurance on the tasks and practice of psychiatry.
Psychiatry, like all professions, is strongly affected by changes in societal expectations and economic forces. Changes in professional priorities and patterns of patient care will undoubtedly be brought about by national health insurance. Two major types of national health insurance have been proposed: comprehensive health insurance and catastrophic insurance. We do not anticipate major impact on psychiatric tasks from some form of catastrophic insurance. Comprehensive health insurance would shape and change psychiatric practice. An examination of psychiatric tasks provides a framework for anticipating alterations in practice. We identify four major task areas in psychiatry: (1) medical tasks, (2) reparative tasks, (3) social control tasks, and (4) humanistic tasks. These tasks would be differentially influenced. Psychiatry's medical tasks will be stressed, while funding for many reparative tasks may be limited. The care of the severely ill patient may be fragmented because of problems in integrating medical and rehabilitative services.


Exchanging partially hydrogenated fat for palmitic acid in the diet increases LDL-cholesterol and endogenous cholesterol synthesis in normocholesterolemic women.
Partial hydrogenation of oil results in fats containing unusual isomeric fatty acids characterized by cis and trans configurations. Hydrogenated fats containing trans fatty acids increase plasma total cholesterol (TC) and LDL-cholesterol while depressing HDL-cholesterol levels. Identifying the content of trans fatty acids by food labeling is overshadowed by a reluctance of health authorities to label saturates and trans fatty acids separately. Thus, it is pertinent to compare the effects of trans to saturated fatty acids using stable isotope methodology to establish if the mechanism of increase in TC and LDL-cholesterol is due to the increase in the rate of endogenous synthesis of cholesterol. Ten healthy normocholesterolemic female subjects consumed each of two diets containing approximately 30% of energy as fat for a fourweek period. One diet was high in palmitic acid (10.6% of energy) from palm olein and the other diet exchanged 5.6% of energy as partially hydrogenated fat for palmitic acid. This fat blend resulted in monounsaturated fatty acids decreasing by 4.9 % and polyunsaturated fats increasing by 2.7%. The hydrogenated fat diet treatment provided 3.1% of energy as elaidic acid. For each dietary treatment, the fractional synthesis rates for cholesterol were measured using deuterium-labeling procedures and blood samples were obtained for blood lipid and lipoprotein measurements. Subjects exhibited a higher total cholesterol and LDL-cholesterol level when consuming the diet containing trans fatty acids while also depressing the HDL-cholesterol level. Consuming the partially hydrogenated fat diet treatment increased the fractional synthesis rate of free cholesterol. Consumption of hydrogenated fats containing trans fatty acids in comparison to a mixtur e of palmitic and oleic acids increase plasma cholesterol levels apparently by increasing endogenous synthesis of cholesterol.


Effect of Ganoderma lucidum on postherpetic neuralgia.
Administration of hot water soluble extracts of Ganoderma lucidum (GI) (36 to 72 g dry weight/day) decreased pain dramatically in two patients with postherpetic neuralgia recalcitrant to standard therapy and two other patients with severe pain due to herpes zoster infection.


The Role of Pain Catastrophizing and Pain Acceptance in Performance-Based and Self-Reported Physical Functioning in Individuals with Fibromyalgia and Obesity.
Impaired physical functioning is one of the most critical consequences associated with fibromyalgia, especially when there is comorbid obesity. Psychological factors are known to contribute to perceived (i.e., subjective) physical functioning. However, physical function is a multidimensional concept encompassing both subjective and objective functioning. The contribution of psychological factors to performance-based (i.e., objective) functioning is unclear. This study aims to investigate the contribution of pain catastrophizing and pain acceptance to both self-reported and performance-based physical functioning. In this cross-sectional study, 160 participants completed self-report measures of pain catastrophizing, pain acceptance, and pain severity. A self-report measure and a performance-based test were used to assess physical functioning. Higher pain catastrophizing and lower pain acceptance were associated with poorer physical functioning at both self-reported and performance-based levels. Our results are consistent with previous evidence on the association between pain catastrophizing and pain acceptance with self-reported physical functioning. This study contributes to the current literature by providing novel insights into the role of psychological factors in performance-based physical functioning. Multidisciplinary interventions that address pain catastrophizing and pain acceptance are recommended and might be effective to improve both perceived and performance-based functioning in women with FM and obesity.


[Derivative of bleomycin generates less of ROS? Less of fibrosis? Alternative in the development of an efficacy anticancer therapy but less toxic].
Deglycobleomycin (DBLM), the aglycon of the glycopeptide antitumor drug bleomycin (BLM), was first used since 1980 during comparative studies between BLM and DBLM in order to elucidate the role of the sugar component in the mechanism of action of BLM. In fact, the deglycosylation of BLM reduce the toxicity of this molecule and fails to produce reactive oxygen species, responsible for pulmonary fibrosis, and for anti-neoplastic activity of BLM. This causes toxic DNA lesions and ultimately leads to cell death. The therapeutic use of BLM is limited by a dose-dependent lung toxicity that eventually leads to fibrosis. Testing BLM-derivative molecules and defining their molecular mechanisms involved in tumor cell death may help to design new therapeutic approach with limited toxicity profile while maintaining anti-tumoral properties. The present review was undertaken to determine the effect of carbohydrate moiety in the mechanism of BLM induced cytotoxicity behind a comparative studies between BLM and DBLM.


Acute Alcohol Consumption Directly Increases HIV Transmission Risk: A Randomized Controlled Experiment.
Alcohol consumption has frequently been purported as a driver of condomless sex and HIV transmission, but to date, experimental evidence for the causal risk-taking impact of alcohol among HIV-positive populations is lacking. The present experiment sought to determine whether acute alcohol consumption has a direct causal impact on condomless sex intentions among HIV-positive men-who-have-sex-with-men (MSM), and to assess whether alcohol's impact differs between MSM who are HIV-positive versus HIV-negative. In a randomized controlled alcohol administration experiment, HIV-positive and HIV-negative MSM were brought into a specialized barroom laboratory and randomly assigned to beverage consumption condition: alcohol (target blood alcohol concentration = 0.080%), placebo alcohol (target blood alcohol concentration = 0.000%), or water (control). Participants then underwent a video-based sexual arousal manipulation (sexually aroused/nonaroused) and indicated their intentions to engage in condom-protected and condomless sexual acts in a standardized paradigm. The primary outcome entailed intentions to engage in condomless receptive and condomless insertive anal sex. A total of 282 MSM (141 HIV-positive; 141 HIV-negative) completed experimental procedures. MSM who received alcohol reported significantly stronger intentions to engage in condomless sex than those who received placebo alcohol or water (F(1,274) = 9.43, P = 0.002). The impact of alcohol did not differ between HIV-positive and HIV-negative MSM (F(1,274) = 1.86, P = 0.174). The present investigation entailed the first risk-focused alcohol administration experiment to involve an HIV-positive sample, and results demonstrated that consuming alcohol had an independent, causal impact on intentions to engage in sexual behaviors that can result in HIV transmission. Findings strongly suggest that alcohol-focused initiatives should be incorporated into HIV prevention efforts.


[Osteosclerotic myeloma. Report on three cases and review of the literature (author's transl)].
Three cases of osteosclerotic myeloma are reported. Clues for diagnosis are agarose gel electrophoresis and sclerotic bone biopsy. Review of 50 cases of literature points out frequency of peripheral neuropathy and Ig A type Immunoglobulin. Iliac crest biopsy shows thickening of bone structures and plasmocytosis in medullary spaces. Main X-Ray signs are mentioned and pathogenesis is discussed.


Genome-wide association study reveals susceptibility loci for self-reported headache in a large community-based Asian population.
The genetic substrate for headache in the general population has not been identified in Asians. We investigated susceptible genetic variants for self-reported headache in a large community-based Asian population. We conducted a genome-wide association study in participants recruited from a community-based cohort to identify the genetic variants associated with headache in Taiwanese. All participants received a structured questionnaire for self-reported headache. A total of 2084 patients with "self-reported headache" and 11,822 age- and sex-matched controls were enrolled. Gene enrichment analysis using the Genotype-Tissue Expression version 6 database was performed to explore the potential function of the identified variants. We identified two novel loci, rs10493859 in TGFBR3 and rs13312779 in FGF23, that are functionally relevant to vascular function and migraine to be significantly associated with self-reported headache after adjusting age, sex and top 10 principal components (p = 8.53 × 10-11 and p = 1.07 × 10-8, respectively). Gene enrichment analysis for genes with GWAS suggestive significance (p < 10-6) demonstrated that the expression of these genes was significantly enriched in the artery (p = 8.18 × 10-4) and adipose tissue (p = 8.95 × 10-4). Our results suggest that vascular dysfunction might play important roles in the pathogenesis of self-reported headache in Asian populations.


Induction of heat shock protein synthesis in murine tumors during the development of thermotolerance.
The function of one or more heat shock proteins (HSPs) may be to confer protection of cells against thermal damage. We examined the induction kinetics of thermotolerance and the synthesis of HSPs in murine tumor models. Squamous cell carcinomas (SCC VII/SF) or radiation-induced fibrosarcomas (RIF) were implanted in the flanks of C3H mice. These flank tumors were first exposed to an elevated temperature (41 degrees-45 degrees C) for a fixed duration, for example, 43 degrees C for 15 min. Some of the tumors were excised immediately, and tumor cell suspensions were made. The other mice with tumors were returned to the cages and left undisturbed for various times up to 72 h before being sacrificed. Again, tumors were then removed and tumor cell suspensions were prepared. These tumor cells were either challenged with a second heat treatment at 45 degrees C in vitro or labeled with [35S]methionine at 37 degrees C in vitro. The tumor cell survival after the combined heat treatments was measured using the in vitro cloning assay. The cellular proteins were analyzed by one- or two-dimensional gel electrophoresis. We found that mild heat shock induced thermotolerance in murine tumors, a result consistent with those of others. The kinetics of induction and decay of thermotolerance depended on the temperature and duration of the priming treatment. Mild heat shock also enhanced the rate of synthesis and accumulation of some HSPs during the development of thermotolerance. For example, after an initial treatment at 43 degrees C for 15 min, the rates of synthesis of HSPs with molecular weights 68,000, 70,000, and 88,000 were greatly enhanced in SCC VII/SF tumors when compared to unheated controls. Qualitatively similar results were seen with radiation-induced fibrosarcoma tumors. The rate of synthesis of Mr 68,000 to 70,000 HSPs reached maximum value (300% of control value) 2 to 4 h after heat shock and decreased to the control value 6 to 24 h later. On the other hand, the rate of synthesis of actin, a major structural cellular protein, remained relatively constant throughout the 72 h of experiments. We then determined the relationship between the synthesis and accumulation of these HSPs and the expression of thermotolerance in murine tumors after a priming heat treatment. The data indicate that the levels of Mr 68,000 to 70,000 HSPs correlate well with thermotolerance.(ABSTRACT TRUNCATED AT 400 WORDS)


Extracellular vesicle-mediated transfer of long non-coding RNA ROR modulates chemosensitivity in human hepatocellular cancer.
Hepatocellular cancers (HCC) are highly resistant to chemotherapy. TGFβ has been associated with chemoresistance in some human cancers but the mechanisms involved are unknown. We explored how TGFβ might contribute to altered responses to therapy by assessing the involvement and mechanistic contribution of extracellular vesicle long non-coding RNA (lncRNA) in mediating TGFβ-dependent chemoresistance. TGFβ reduced the sensitivity of HCC cells to sorafenib or doxorubicin and altered the release of both extracellular vesicles and of selected lncRNA within these vesicles. Amongst these, lincRNA-ROR (linc-ROR), a stress-responsive lncRNA was highly expressed in HCC cells and enriched within extracellular vesicles derived from tumor cells. Incubation with HCC-derived extracellular vesicles increased linc-ROR expression and reduced chemotherapy-induced cell death in recipient cells. Sorafenib increased linc-ROR expression in both tumor cells and extracellular vesicles, whereas siRNA to linc-ROR increased chemotherapy-induced apoptosis and cytotoxicity. Tumor-initiating cells that express CD133 have an increased resistance to therapy. TGFβ increased expression of CD133+ cells and colony growth in limiting dilution assays, both of which were attenuated by linc-ROR knockdown. These data provide mechanistic insights into primary chemoresistance in HCC by showing that: (a) TGFβ selectively enriches linc-RoR within extracellular vesicles, which has a potential role in intercellular signaling in response to TGFβ; (b) expression and enrichment of linc-ROR during chemotherapeutic stress plays a functional role in chemoresistance; and (c) the effects of TGFβ on chemoresistance in HCC may involve linc-RoR-dependent effects on tumor-initiating cells. These findings implicate extracellular vesicle lncRNA as mediators of the chemotherapeutic response, and support targeting linc-ROR to enhance chemosensitivity in HCC.


Attenuated influenza virions expressing the SARS-CoV-2 receptor-binding domain induce neutralizing antibodies in mice.
An effective vaccine is essential to controlling the spread of SARS-CoV-2 virus. Here, we describe an influenza-virus-based vaccine for SARS-CoV-2. We incorporated a membrane-anchored form of the SARS-CoV-2 Spike receptor binding domain (RBD) in place of the neuraminidase (NA) coding sequence in an influenza virus also possessing a mutation that reduces the affinity of hemagglutinin for its sialic acid receptor. The resulting ΔNA(RBD)-Flu virus can be generated by reverse genetics and grown to high titers in cell culture. A single-dose intranasal inoculation of mice with ΔNA(RBD)-Flu elicits serum neutralizing antibody titers against SAR-CoV-2 comparable to those observed in humans following natural infection (~1:200). Furthermore, ΔNA(RBD)-Flu itself causes no apparent disease in mice. It might be possible to produce a vaccine similar to ΔNA(RBD)-Flu at scale by leveraging existing platforms for production of influenza vaccines.


Preemptive intrathecal ketamine delays mechanical hyperalgesia in the neuropathic rat.
We studied the effect of preemptive intrathecal ketamine administration on the development of mechanical hyperalgesia in a rat model of mononeuropathy. Rats given intrathecal ketamine 1 mg/kg or normal saline were subsequently rendered neuropathic by placing four loose ligatures around the sciatic nerve. The onset of resultant hyperalgesia was evaluated using von Frey monofilaments. Preemptive ketamine significantly slowed the onset of mechanical hyperalgesia at both 2.35 g (P < 0.04) and 4.19 g (P < 0.05) thresholds through the third postoperative day but did not alter the ultimate degree of hyperalgesia in either severity or frequency in this model. We conclude that ketamine delayed the onset, but failed to prevent the development of, mechanical hyperalgesia in a rat model of peripheral mononeuropathy. Pain in response to light touch may follow nerve injury. Prevention of this complication would be clinically useful. Ketamine, an anesthetic, was administered spinally to rats before nerve injury in an attempt to prevent the development of chronic pain. Hypersensitivity to light touch was delayed by preemptive ketamine.


A Target Capture-Based Method to Estimate Ploidy From Herbarium Specimens.
Whole genome duplication (WGD) events are common in many plant lineages, but the ploidy status and possible occurrence of intraspecific ploidy variation are unknown for most species. Standard methods for ploidy determination are chromosome counting and flow cytometry approaches. While flow cytometry approaches typically use fresh tissue, an increasing number of studies have shown that recently dried specimens can be used to yield ploidy data. Recent studies have started to explore whether high-throughput sequencing (HTS) data can be used to assess ploidy levels by analyzing allelic frequencies from single copy nuclear genes. Here, we compare different approaches using a range of yam (Dioscorea) tissues of varying ages, drying methods and quality, including herbarium tissue. Our aims were to: (1) explore the limits of flow cytometry in estimating ploidy level from dried samples, including herbarium vouchers collected between 1831 and 2011, and (2) optimize a HTS-based method to estimate ploidy by considering allelic frequencies from nuclear genes obtained using a target-capture method. We show that, although flow cytometry can be used to estimate ploidy levels from herbarium specimens collected up to fifteen years ago, success rate is low (5.9%). We validated our HTS-based estimates of ploidy using 260 genes by benchmarking with dried samples of species of known ploidy (Dioscorea alata, D. communis, and D. sylvatica). Subsequently, we successfully applied the method to the 85 herbarium samples analyzed with flow cytometry, and successfully provided results for 91.7% of them, comprising species across the phylogenetic tree of Dioscorea. We also explored the limits of using this HTS-based approach for identifying high ploidy levels in herbarium material and the effects of heterozygosity and sequence coverage. Overall, we demonstrated that ploidy diversity within and between species may be ascertained from historical collections, allowing the determination of polyploidization events from samples collected up to two centuries ago. This approach has the potential to provide insights into the drivers and dynamics of ploidy level changes during plant evolution and crop domestication.


Transient Measurements and Simulations Correlate Exchange Ligand Concentration and Trap States in Colloidal Quantum Dot Photodetectors.
Colloidal quantum dot (CQD) photodetectors (PDs) can detect wavelengths longer than the 1100 nm limit of silicon because of their highly tunable bandgaps. CQD PDs are acutely affected by the ligands that separate adjacent dots in a CQD-solid. Optimizing the exchange solution ligand concentration in the processing steps is crucial to achieving high photodetector performance. However, the complex mix of chemistry and optoelectronics involved in CQD PDs means that the effects of the exchange solution ligand concentration on device physics are poorly understood. Here we report direct correspondence between simulated and experimental transient photocurrent responses in CQD PDs. For both deficient and excess conditions, our model demonstrated the experimental changes to the transient photocurrent aligned with changes in trap state density. Combining transient photoluminescence, absorption, and photocurrent with this simulation model, we revealed that different mechanisms are responsible for the increased trap density induced by excess and deficient active layer ligand concentrations.


CHOROIDAL NEVUS ASSOCIATED WITH VITELLIFORM DEPOSITION IN A PATIENT WITH AUTOSOMAL DOMINANT BEST DYSTROPHY.
To describe the clinical, optical coherence tomography (OCT), fundus autofluorescence and ultrasound findings of a patient with a choroidal nevus actively exuding vitelliform material in the setting of autosomal dominant Best dystrophy (BD). The patient's clinical course was followed over time with ophthalmic examinations and multimodal imaging. A 71-year-old male patient with BD was referred for evaluation of a choroidal nevus in the right eye. Dilated fundoscopic examination showed a small pigmented choroidal nevus in the temporal periphery. Over a 3-year period, the nevus developed progressive deposition of vitelliform material along its inferior border. Meanwhile, OCT and fundus photography showed only slight growth. Ultrasound showed no change in height; basal measurements were confounded by the increased vitelliform deposits. Genetic testing confirmed a heterozygous mutation in the BEST1 gene and electrophysiology was consistent with BD. Dysfunction of the retinal pigment epithelium associated with BD may cause novel presentations of other conditions such as choroidal nevi. The implication for malignant transformation of a choroidal nevus associated with vitelliform deposit accumulation in this context is unknown.


Changes in the temperature sensitivity of SOM decomposition with grassland succession: implications for soil C sequestration.
Understanding the temperature sensitivity (Q 10) of soil organic matter (SOM) decomposition is important for predicting soil carbon (C) sequestration in terrestrial ecosystems under warming scenarios. Whether Q 10 varies predictably with ecosystem succession and the ways in which the stoichiometry of input SOM influences Q 10 remain largely unknown. We investigate these issues using a grassland succession series from free-grazing to 31-year grazing-exclusion grasslands in Inner Mongolia, and an incubation experiment performed at six temperatures (0, 5, 10, 15, 20, and 25°C) and with four substrates: control (CK), glucose (GLU), mixed grass leaf (GRA), and Medicago falcata leaf (MED). The results showed that basal soil respiration (20°C) and microbial biomass C (MBC) logarithmically decreased with grassland succession. Q 10 decreased logarithmically from 1.43 in free-grazing grasslands to 1.22 in 31-year grazing-exclusion grasslands. Q 10 increased significantly with the addition of substrates, and the Q 10 levels increased with increase in N:C ratios of substrate. Moreover, accumulated C mineralization was controlled by the N:C ratio of newly input SOM and by incubation temperature. Changes in Q 10 with grassland ecosystem succession are controlled by the stoichiometry of newly input SOM, MBC, and SOM quality, and the combined effects of which could partially explain the mechanisms underlying soil C sequestration in the long-term grazing-exclusion grasslands in Inner Mongolia, China. The findings highlight the effect of substrate stoichiometry on Q 10 which requires further study.