HIV protease inhibitors: immunological insights.
Regimens of highly active antiretroviral therapy (HAART), including those containing one or more of the potent HIV protease inhibitors, often cause large increases in CD4 cell count as well as large decreases in plasma HIV RNA that persist over time. Patients improve clinically, and the occurrence (or recurrence) of opportunistic diseases appears to be reduced. This raises issues about the source and functional capacity of the CD4 cells seen in patients receiving HAART. This article briefly summarizes the current information about the recirculation of T cells from lymphoid tissue to blood after HAART, increases in the number of naive (CD45RA+, CD62L+) and memory (CD45RO+) CD4 cells, changes in the T cell repertoire (Vbeta), and the possible role of selected cytokines that may be useful in facilitating immune reconstitution in patients receiving HAART. Preliminary data from some studies have revealed that, after several months of therapy, the number of memory CD4 cells increased, followed by increases in naive CD4 cells. Other studies have found that the number of naive CD4 cells increases only if these cells were present before initiation of therapy and that disruptions of the Vbeta subsets are not immediately corrected. Studies concerning the relations of various cytokines to immune reconstitution are also ongoing. On the basis of the limited results available in late 1997, there is good reason to hope for at least partial immune reconstitution in HIV patients treated with HAART, especially if therapy is initiated before severe damage to the immune system has occurred. In the future, cytokines such as interleukin-2, vaccinations, cellular replacement or stem cell transfer, or bone marrow transplants may play important roles in therapy.


The Chondrogenic Potential of Mesenchymal Cells and Chondrocytes from Osteoarthritic Subjects: A Comparative Analysis.
The multipotential nature of stem or progenitor cells apparently makes them the ideal choice for any cell therapy, but this as yet remains to be proven. This study (30 subjects) was designed to compare the potential to repair articular cartilage of chondrocytes taken from different regions in osteoarthritic cartilage with that of mesenchymal stem cells prepared from bone marrow of the same subject. Cartilage biopsies, bone marrow, and blood samples were taken from each of 30 individuals with chronic osteoarthritis (aged 62-85 years) undergoing total knee replacement. The chondrogenic potential of chondrocytes isolated from cartilage biopsies taken from different regions of osteoarthritic cartilage was compared with that of mesenchymal cells by quantitative analysis of several chondrocyte specific markers and an ex vivo cartilage differentiation assay. Cartilage-derived articular chondrocytes are superior to bone marrow-derived cells when compared for their ex vivo chondrogenic potential. Interestingly, there was marked and significant difference in the expression of chondrocytic markers between chondrocytes derived from adjacent, visually distinct regions of the diseased cartilage. When cultured in the presence of a fibroblast growth factor 2 variant, all cell samples from both tissues showed a high degree of chondrogenic potential. Although bone marrow-derived mesenchymal cells, when supplemented with the appropriate chondrogenic factors, are a suitable source for autologous cartilage implantation, adult chondroprogenitor cells, particularly those from moderately affected regions of the osteoarthritic joints, demonstrate superior chondrogenic potential.


CC chemokine receptor-7 on dendritic cells is induced after interaction with apoptotic tumor cells: critical role in migration from the tumor site to draining lymph nodes.
Dendritic cells (DCs) are very potent antigen-presenting cells and play critical roles in regulating immune responses in cancer. The migrating of DCs from the tumor site to the lymphoid organs is believed to be one of the critical events. To examine this important DC function in tumor situations, bone marrow-derived DCs, cultured for 6 days with granulocyte macrophage colony-stimulating factor and interleukin 4, were inoculated at the tumor site. We have shown (Y. Nishioka et al., Cancer Res., 59: 40354041, 1999) that DCs can migrate from tumor site to the draining lymph nodes within 24 h (approximately 0.1% of administrated DCs). The DCs then form clusters with adjacent lymphoid cells, which produce IFN-gamma (1500-3200pg/10(6) cells/48 h) in response to tumor stimulation. The number of the DCs migrating into lymph nodes were greater when they were inoculated into the tumor rather than the skin. Coculture of DCs and apoptotic tumor cells resulted in decreased expression of CC chemokine receptor (CCR) 1 and increased CCR7 expression at mRNA level without alteration in other phenotypical markers on DCs. Chemotaxis assay showed that CCR7 ligands, macrophage inflammatory protein 3beta and secondary lymphoid-tissue chemokine significantly (P < 0.05) induced the migration of DCs when cocultured with apoptotic tumor cells. To directly examine the involvement of CCR7 expression in DC migration, we investigated the functions of DCs genetically modified to express high levels of CCR7. CCR7 transduction promotes DC migration in response to relevant ligands in vitro and in vivo. These results suggest that the CCR7 expression of DCs is enhanced with direct contact with apoptotic tumor cells and may have a critical role for DC migrating to regional lymph nodes. The means to promote DC delivery to tumor and to nodal sites represent novel targets for the biological therapy of cancer.


[Perioperative management of a patient with deep vein thrombosis caused by estrogen producing granulosa cell tumor of the ovary].
We experienced perioperative management of a woman with large thrombi in femoral veins caused by estrogen producing ovarian tumor. At presentation, serum estradiol level was extremely high. Time constraint due to rapid expansion of the tumor did not allow us a trial of preoperative anti-coagulation. We placed a temporary IVC filter and proceeded with operation. As catastrophic pulmonary embolism could occur even with the presence of a filter, we prepared for institution of cardiopulmonary bypsss and pulmonary embolectomy via full sternotomy. We monitored arterial pressure based-cardiac output for immediate detection of pulmonary embolism. Postoperatively, serum estradiol level declined to normal range by postoperative day 7, and thrombi dissolved with anticoagulation therapy with warfarin and heparin without recurrence. In this case, surgery without preoperative anticoagulation was considered to pose extreme risk of perioperative pulmonary embolism. However, we considered that the thrombi would disolve by the removal of the tumor because estradiol production was the primary cause of thrombi. With above mentioned perioperative approach, the patient was managed uneventfully.


[Role of the social support network in caring for the aged].
This study presents the results of an investigation of the reasons for the elderly people being cared for as residents of nursing homes or as outpatients. With regard to the reasons for being outpatients or being residents of nursing homes it can be pointed out: elderly people as outpatients can only be cared for as long as there are intact social relations. If services for outpatients are not supplemented by social relations the stay in a nursing home cannot be avoided.


Uric acid and HMGB1 are involved in the induction of autoantibodies elicited in mice infected with mouse hepatitis virus A59.
We have shown that mice infected with mouse hepatitis virus A59 develop autoantibodies (autoAb) to liver and kidney fumarylacetoacetate hydrolase (FAH). Because it has been proposed that the immune system is stimulated by alarm signals called damage-associated molecular patterns or alarmins, we investigated the participation of uric acid and high-mobility group box protein 1 (HMGB1) in the autoimmune response elicited by mouse hepatitis virus (MHV). Mice subjected to MHV infection had increased plasmatic uric acid concentration that significantly decreased after 20 days of daily treatment with allopurinol and, simultaneously, autoAb to FAH were undetected. Furthermore, this autoAb disappeared after 30 days of treatment with ethyl pyruvate, along with a substantial reduction in serum HMGB1 concentration. Both results indicated a remarkable relationship between the autoimmune process induced by the virus and uric acid and HMGB1 liberation. Unexpectedly, it was found that allopurinol and ethyl pyruvate inhibited the release of both uric acid and HMGB1. Because HMGB1 is activated through binding to interleukin 1β, and that this cytokine is produced by the NLRP3 inflammasome that could be stimulated by uric acid, we propose that both alarmins could be acting in concert with the induction of the autoAb to FAH in MHV-infected mice.


Arteriovenous malformations of the mandible. Graduated surgical management.
Two cases of arteriovenous malformation in the mandible are described, in which muscle embolization was used. The success in one patient and the failure in the second correlate with the caliber of the vessels in the two malformations. A scheme of graduated surgical management for mandibular arteriovenous malformations is proposed.


Regulation of gut and heart left-right asymmetry by context-dependent interactions between xenopus lefty and BMP4 signaling.
The Lefty subfamily of TGFbeta signaling molecules has been implicated in early development in mouse, zebrafish, and chick. Here, we show that Xenopus lefty (Xlefty) is expressed both bilaterally in symmetric midline domains and unilaterally in left lateral plate mesoderm and anterior dorsal endoderm. To examine the roles of Xlefty in left-right development, we created a system for scoring gut asymmetry and examined the effects of unilateral Xlefty misexpression on gut development, heart development, and Xnr-1 and XPitx2 expression. In contrast to the unilateral effects of Vg1, Activin, Nodal, or BMPs, targeted expression of Xlefty in either the left or the right side of Xenopus embryos randomized the direction of heart looping, gut coiling, and left-right positioning of the gut and downregulated the asymmetric expression of Xnr-1 and XPitx2. It is currently thought that Lefty proteins act as feedback inhibitors of Nodal signaling. However, this would not explain the effects of right-sided Xlefty misexpression. Here, we show that Xlefty interacts with the signaling pathways of other members of the TGFbeta family during left-right development. Results from coexpression of Xlefty and Vg1 indicate that Xlefty can nullify the effects of Vg1 ectopic expression and that Xlefty is downstream of left-sided Vg1 signaling. Results from coexpression of Xlefty and XBMP4 indicate that XLefty and XBMP4 interact both synergistically and antagonistically in a context-dependent manner. We propose a model in which interactions of Xlefty with multiple members of the TGFbeta family enhance the differences between the right-sided BMP/ALK2/Smad pathway and the left-sided Vg1/anti-BMP/Nodal pathway, leading to left-right morphogenesis of the gut and heart.


The crucial role of plant mitochondria in orchestrating drought tolerance.
Around the world, the frequency and intensity of droughts is increasing as a result of global climate change, with important consequences for the growth and survival of agricultural and native plant species. Understanding how plants respond to water stress is thus crucial for predicting the impacts of climate change on the crop productivity and ecosystem functioning. In contrast to the large number of studies assessing drought impacts on photosynthesis, relatively little attention has been devoted to understanding how mitochondrial respiratory metabolism is altered under water stress conditions. This review provides an overview of the impacts of water stress on mitochondrial respiration (R), combining studies at the whole-plant, individual organ, cellular and organelle levels. To establish whether there are clear patterns in the response of in vivo R to water stress, a wide range of root, leaf and whole-plant studies are reviewed. It is shown that water stress almost always inhibits R in actively growing roots and whole plants. However, in fully expanded, mature leaves the response is more variable, with water stress reducing R in near two-thirds of reported studies, with most of the remainder showing no change. Only a few studies reported increases in leaf R under severe water stress conditions. The mechanisms responsible for these variable responses are discussed. Importantly, the fact is highlighted that irrespective of whether drought increases or decreases respiration, overall the changes in R are minor compared with the large decreases in photosynthetic carbon gain in response to drought. Based on recent work highlighting the link between chloroplast and mitochondrial functions in leaves, we propose a model by which mitochondrial R enables survival and rapid recovery of productivity under water stress conditions. Finally, the effects of water stress on mitochondrial function, protein abundance and overall metabolism are reviewed.


Trypanosomatidae from wild mammals in the neotropical rainforest of French Guiana.
The initial filling of the reservoir behind the Petit Saut hydro-electric dam, on the Sinnamary River in French Guiana, threatened the terrestrial and arboreal animals living in the neotropical rainforest being flooded. During a rescue programme between 24 October and 12 November in 1994, many of these animals were checked for infection with trypanosomatids. Overall, 45 blood samples and 54 skin biopsies were collected from 53 mammals (of 13 species representing five orders) and blood samples were also taken from each of nine reptiles (six species from four families). When the skin biopsies and the buffy-coats from the blood samples were cultured in NNN medium, 10 of the cultures, each initiated with mammalian blood, were found to be positive for trypanosomatids. Multilocus enzyme electrophoresis (MLEE) on cellulose acetate plates, with 20 enzyme systems, was then used to investigate each of the positive cultures. The results were analysed by clustering from a genetic distance matrix, using the unweighted pair group method with arithmetic averages (UPGMA), and applying a bootstrap procedure to Wagner parsimony trees. A stock obtained from Didelphis marsupialis was identified as a zymodeme of Trypanosoma cruzi (Miles' zymodeme 1) known to cause Chagas disease in French Guiana. Five stocks (one each from Bradypus tridactylus, Tamandua tetradactyla and Alouatta seniculus and two from Saguinus midas) were of a single zymodeme close to Trypanosoma rangeli reference stock RGB. This is the first confirmation of the presence of Tr. rangeli in French Guiana, and the first time that it has been identified, by iso-enzyme analysis, in the neotropical primates A. seniculus and S. midas. Two other stocks, isolated from Choloepus didactylus, were related to Endotrypanum schaudinni reference stock LEM 2790. Although the remaining stocks, one from C. didactylus and the other from A. seniculus, clustered together on UPGMA and in a Wagner tree, they did not appear to be related to any of the reference stocks included in the UPGMA dendrogram.


Complications of tibial eminence and diaphyseal fractures in children: prevention and treatment.
Fractures of the tibial eminence and the diaphyseal tibia are common pediatric orthopaedic injuries. Although most tibial fractures can be treated nonsurgically, those that require surgical intervention may encounter specific complications. Surgical treatment of fractures of the tibial eminence may be complicated by failed fixation, knee joint stiffness, and arthrofibrosis of the knee, a complication rarely seen in children but occurring most frequently after tibial eminence injuries. Complications of healing after tibial fractures in pediatric patients are uncommon, although some tibial shaft fractures exhibit delayed union or nonunion, infection, and soft-tissue complications.


Highly Polyvalent DNA Motors Generate 100+ pN of Force via Autochemophoresis.
Motor proteins such as myosin, kinesin, and dynein are essential to eukaryotic life and power countless processes including muscle contraction, wound closure, cargo transport, and cell division. The design of synthetic nanomachines that can reproduce the functions of these motors is a longstanding goal in the field of nanotechnology. DNA walkers, which are programmed to "walk" along defined tracks via the burnt bridge Brownian ratchet mechanism, are among the most promising synthetic mimics of these motor proteins. While these DNA-based motors can perform useful tasks such as cargo transport, they have not been shown to be capable of cooperating to generate large collective forces for tasks akin to muscle contraction. In this work, we demonstrate that highly polyvalent DNA motors (HPDMs), which can be viewed as cooperative teams of thousands of DNA walkers attached to a microsphere, can generate and sustain substantial forces in the 100+ pN regime. Specifically, we show that HPDMs can generate forces that can unzip and shear DNA duplexes (∼12 and ∼50 pN, respectively) and rupture biotin-streptavidin bonds (∼100-150 pN). To help explain these results, we present a variant of the burnt-bridge Brownian ratchet mechanism that we term autochemophoresis, wherein many individual force generating units generate a self-propagating chemomechanical gradient that produces large collective forces. In addition, we demonstrate the potential of this work to impact future engineering applications by harnessing HPDM autochemophoresis to deposit "molecular ink" via mechanical bond rupture. This work expands the capabilities of synthetic DNA motors to mimic the force-generating functions of biological motors. Our work also builds upon previous observations of autochemophoresis in bacterial transport processes, indicating that autochemophoresis may be a fundamental mechanism of pN-scale force generation in living systems.


A Case of Vitiligo after Kawasaki's Disease.
Vitiligo is a common skin disease, but its pathogenesis has not been fully determined, though an autoimmune etiology is considered likely. Kawasaki disease (KD) is an acute multisystem vasculitis of childhood associated with coronary arteriopathy, and is diagnosed based on clinical criteria. Furthermore, vitiligo has been associated with several other diseases, but no report has been issued about the relationship between vitiligo and Kawasaki's disease. The author's report the case of an 8-year-old male child that presented with depigmented lesions, which developed from the desquamative skin lesions of Kawasaki's disease.


Comparative analysis of storage conditions and homogenization methods for tick and flea species for identification by MALDI-TOF MS.
Ticks and fleas are vectors for numerous human and animal pathogens. Controlling them, which is important in combating such diseases, requires accurate identification, to distinguish between vector and non-vector species. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was applied to the rapid identification of arthropods. The growth of this promising tool, however, requires guidelines to be established. To this end, standardization protocols were applied to species of Rhipicephalus sanguineus (Ixodida: Ixodidae) Latreille and Ctenocephalides felis felis (Siphonaptera: Pulicidae) Bouché, including the automation of sample homogenization using two homogenizer devices, and varied sample preservation modes for a period of 1-6 months. The MS spectra were then compared with those obtained from manual pestle grinding, the standard homogenization method. Both automated methods generated intense, reproducible MS spectra from fresh specimens. Frozen storage methods appeared to represent the best preservation mode, for up to 6 months, while storage in ethanol is also possible, with some caveats for tick specimens. Carnoy's buffer, however, was shown to be less compatible with MS analysis for the purpose of identifying ticks or fleas. These standard protocols for MALDI-TOF MS arthropod identification should be complemented by additional MS spectrum quality controls, to generalize their use in monitoring arthropods of medical interest.


Depth and shape from differential perspective in the presence of bending deformations.
It is shown that there exist classes of useful partial solutions to what has become known as the structure from motion problem, that is, the problem of finding the three-dimensional shape as well as the movement in space of objects on the basis of their optical projections in several distinct phases of the movement. The class of solutions treated in this paper is quite different from the usual treatment in that the rigidity condition is relaxed so as to include the important class of bending deformations. For instance, we show a case in which important aspects of shape are obtained for a configuration of seven points, no four of which move in a rigid way, and which can be obtained from only two views. (This violates all requirements of the structure from motion theorem.) We discuss an algorithm that obtains the three-dimensional configuration of a polyhedral vertex up to a relief transformation (a similar transformation to the one that maps object to image space in paraxial optics), in the presence of arbitrary flections of the edges, from the projection and the instantaneous-movement parallax. Such a solution can, for instance, be used to predict side views of the vertex, as is demonstrated for a specific example. The solution can be used in an iterative fashion on triangulations to solve the case of extended curved surfaces suffering arbitrary bendings. The class of bending deformations encompasses all deformations that conserve distances along the surface (but not necessarily through space); thus stretchings are excluded. Practical examples include, e.g., deformations of flexible but nonstretchable shells. Actual numerical examples illustrate the practicability of the method.


Tick ecology: processes and patterns behind the epidemiological risk posed by ixodid ticks as vectors.
The population ecology of ticks is fundamental to the spatial and temporal variation in the risk of infection by tick-borne pathogens. Tick population dynamics can only be fully understood by quantifying the rates of the demographic processes, which are influenced by both abiotic (climatic) factors acting on the free-living tick stages and biotic (host) responses to the tick as a parasite. Within the framework of a population model, I review methods and results of attempts to quantify (1) rates of tick development and the probability of diapause, (2) the probability of questing for hosts by unfed ticks, (3) the probability of ticks attaching to a host, and (4) tick mortality rates. Biologically, these processes involve the physiological and behavioural response of ticks to temperature, moisture stress and day length that result in specific patterns of seasonal population dynamics and host relationships. Temperate and tropical patterns will be illustrated with reference mostly to Ixodes ricinus and Rhipicephalus appendiculatus, respectively.


Patient and Disease Characteristics Associated With the Presence of Diabetes Mellitus in Adults With Chronic Pancreatitis in the United States.
Diabetes mellitus (DM) is a common complication of chronic pancreatitis (CP). Past studies for DM risk factors in CP have been limited to single centers or highly focused on a single etiology such as alcoholic or hereditary disease. We studied risk factors for DM in a large population of patients with CP of all etiologies enrolled in the North American Pancreatitis 2 studies. Participants (1,171) with CP (n=383 with DM, n=788 without DM) were enrolled prospectively from 26 participating centers. Questionnaires were completed by patients and physicians in a cross-sectional assessment. Patient demographics and disease characteristics were compared for CP with DM vs. without DM. Logistic regression was performed to assess the variables associated with DM diagnosis in a multivariable model. Diabetics were more likely to be black (P=0.02), overweight, or obese (P<0.001), and with a family history of DM (P=0.0005). CP patients with DM were more likely to have pancreatic calcifications (63% vs. 54%, P=0.002), atrophy (44% vs. 32%, P<0.0001), and prior pancreas surgery (26.9% vs. 16.9%, P<0.0001). In multivariate logistic regression modeling, the strongest risk factors for DM were obesity (odds ratio (OR) 2.8, 95% confidence interval (CI) 1.9, 4.2) and exocrine insufficiency (OR 2.4, 95% CI 1.8, 3.2). In this large multicenter cohort of patients with CP, exocrine insufficiency, calcifications, and pancreas surgery conveyed higher odds of having DM. However, the traditional 'type 2 DM' risk factors of obesity and family history were similarly important in conveying risk for DM.


Optimizing organic electrosynthesis through controlled voltage dosing and artificial intelligence.
Organic electrosynthesis can transform the chemical industry by introducing electricity-driven processes that are more energy efficient and that can be easily integrated with renewable energy sources. However, their deployment is severely hindered by the difficulties of controlling selectivity and achieving a large energy conversion efficiency at high current density due to the low solubility of organic reactants in practical electrolytes. This control can be improved by carefully balancing the mass transport processes and electrocatalytic reaction rates at the electrode diffusion layer through pulsed electrochemical methods. In this study, we explore these methods in the context of the electrosynthesis of adiponitrile (ADN), the largest organic electrochemical process in industry. Systematically exploring voltage pulses in the timescale between 5 and 150 ms led to a 20% increase in production of ADN and a 250% increase in relative selectivity with respect to the state-of-the-art constant voltage process. Moreover, combining this systematic experimental investigation with artificial intelligence (AI) tools allowed us to rapidly discover drastically improved electrosynthetic conditions, reaching improvements of 30 and 325% in ADN production rates and selectivity, respectively. This powerful AI-enhanced experimental approach represents a paradigm shift in the design of electrified chemical transformations, which can accelerate the deployment of more sustainable electrochemical manufacturing processes.


Erratum for the Report "Reaction of O2 with subsurface oxygen vacancies on TiO2 anatase (101)" by M. Setvín, U. Aschauer, P. Scheiber, Y.-F. Li, W. Hou, M. Schmid, A. Selloni, U. Diebold.
In the Report "Reaction of O2 with subsurface oxygen vacancies on TiO2 anatase (101)," new results indicate that the species referred as (O2–)ads is an adsorbed water molecule, and the species named (O22–)ads is a terminal OH group. Revisited identification of the species is described in Setvín et al., Phys. Chem. Chem. Phys. 16, 21524 (2014). The field-induced oxygen vacancy migration remains valid as described in the Report; the process is discussed in detail in Setvín et al., Phys. Rev. B 91, 195403 (2015). All theoretical results, notably the pathway in Fig. 1, remain valid as well.


Vitamin E: Regulatory Role on Signal Transduction.
Vitamin E modulates signal transduction pathways by several molecular mechanisms. As a hydrophobic molecule located mainly in membranes it contributes together with other lipids to the physical and structural characteristics such as membrane stability, curvature, fluidity, and the organization into microdomains (lipid rafts). By acting as the main lipid-soluble antioxidant, it protects other lipids such as mono- and poly-unsaturated fatty acids (MUFA and PUFA, respectively) against chemical reactions with reactive oxygen and nitrogen species (ROS and RNS, respectively) and prevents membrane destabilization and cellular dysfunction. In cells, vitamin E affects signaling in redox-dependent and redox-independent molecular mechanisms by influencing the activity of enzymes and receptors involved in modulating specific signal transduction and gene expression pathways. By protecting and preventing depletion of MUFA and PUFA it indirectly enables regulatory effects that are mediated by the numerous lipid mediators derived from these lipids. In recent years, some vitamin E metabolites have been observed to affect signal transduction and gene expression and their relevance for the regulatory function of vitamin E is beginning to be elucidated. In particular, the modulation of the CD36/FAT scavenger receptor/fatty acids transporter by vitamin E may influence many cellular signaling pathways relevant for lipid homeostasis, inflammation, survival/apoptosis, angiogenesis, tumorigenesis, neurodegeneration, and senescence. Thus, vitamin E has an important role in modulating signal transduction and gene expression pathways relevant for its uptake, distribution, metabolism, and molecular action that when impaired affect physiological and patho-physiological cellular functions relevant for the prevention of a number of diseases. © 2018 IUBMB Life, 71(4):456-478, 2019.


[Application of damper craniofixators from nitinol in plasty of skull defects].
An overview of the methods of fixation of the skull bones is presented,as well as discussion and characterization of different methods of fixation. A new system of fixation of the skull bones was used in osseous-plastic trepanation and reconstructive operations using nitinol craniofixators. Results of cranioplasty using craniofixators were analyzed in 62 patients with craniocerebral injuries and its consequences, vascular and oncological diseases of the brain. There were no complications in the postoperative period. Articles made of nitinol are very strong, elastic and have the effect of thermo-mechanical memory of the form.


Integrin-β1 is required for the renal cystogenesis caused by ciliary defects.
Defects in the function of primary cilia are commonly associated with the development of renal cysts. On the other hand, the intact cilium appears to contribute a cystogenic signal whose effectors remain unclear. As integrin-β1 is required for the cystogenesis caused by the deletion of the polycystin 1 gene, we asked whether it would be similarly important in the cystogenetic process caused by other ciliary defects. We addressed this question by investigating the effect of integrin-β1 deletion in a ciliopathy genetic model in which the Ift88 gene, a component of complex B of intraflagellar transport that is required for the proper assembly of cilia, is specifically ablated in principal cells of the collecting ducts. We showed that the renal cystogenesis caused by loss of Ift88 is prevented when integrin-β1 is simultaneously depleted. In parallel, pathogenetic manifestations of the disease, such as increased inflammatory infiltrate and fibrosis, were also significantly reduced. Overall, our data indicate that integrin-β1 is also required for the renal cystogenesis caused by ciliary defects and point to integrin-β1-controlled pathways as common drivers of the disease and as possible targets to interfere with the cystogenesis caused by ciliary defects.


Tall Pinus luzmariae trees with genes from P. herrerae.
Pinus herrerae and P. luzmariae are endemic to western Mexico, where they cover an area of more than 1 million hectares. Pinus herrerae is also cultivated in field trials in South Africa and South America, because of its considerable economic importance as a source of timber and resin. Seed quality, afforestation success and desirable traits may all be influenced by the presence of hybrid trees in seed stands. We aimed to determine the degree of hybridization between P. herrerae and P. luzmariae in seed stands of each species located in the Sierra Madre Occidental, Durango, Mexico. AFLP molecular markers from samples of 171 trees across five populations were analyzed with STRUCTURE and NewHybrids software to determine the degree of introgressive hybridization. The accuracy of STRUCTURE and NewHybrids in detecting hybrids was quantified using the software Hybridlab 1.0. Morphological analysis of 131 samples from two populations of P. herrerae and two populations of P. luzmariae was also conducted by Random Forest classification. The data were compared by Principal Coordinate Analysis (PCoA) in GenAlex 6.501. Hybridization between Pinus herrerae and P. luzmariae was observed in all seed stands under study and resulted in enhancement of desirable silvicultural traits in the latter species. In P. luzmariae, only about 16% molecularly detected hybrids correspond to those identified on a morphological basis. However, the morphology of P. herrerae is not consistent with the molecularly identified hybrids from one population and is only consistent with 3.3 of those from the other population. This is the first report of hybrid vigour (heterosis) in Mexican pines. Information about hybridization and introgression is essential for developing effective future breeding programs, successful establishment of plantations and management of natural forest stands. Understanding how natural hybridization may influence the evolution and adaptation of pines to climate change is a cornerstone to sustainable forest management including adaptive silviculture.


Presence of Antibodies against Severe Fever with Thrombocytopenia Syndrome Virus in Non-Endemic Areas of China.
To determine the seroprevalence of severe fever with thrombocytopenia syndrome (SFTS) virus (SFTSV) and identify potential risk factors in non-endemic areas, a cross-sectional study was conducted among a healthy population without previously reported SFTS in a mountainous area of Anhui Province. We recruited and collected blood samples from 270 participants in 4 age groups. An enzyme-linked immunosorbent assay kit was used to detect immunoglobulin G (IgG) antibodies against SFTSV. The overall seropositive rate of SFTSV-IgG among study participants was 6.3% (17/270). We found a significant difference in the SFTSV seropositivity rate between tea pluckers (9.4%, 14/149) and non-tea pluckers (2.5%, 3/121). An increasing trend in the seropositivity rate was found with age for all participants (1.5% to 14.3%) and tea pluckers (2.8% to 19.4%). The SFTSV seropositivity rate in men was higher than that in women for all participants and tea pluckers, but the differences were not significant. Critical risk factors for SFTSV infection are increasing age and being a tea plucker. Our findings suggest that health education should be enhanced to increase awareness among residents (especially, the older ones and tea pluckers) to protect them against SFTS.


Depth Distribution and Assembly of Sulfate-Reducing Microbial Communities in Marine Sediments of Aarhus Bay.
Most sulfate-reducing microorganisms (SRMs) present in subsurface marine sediments belong to uncultured groups only distantly related to known SRMs, and it remains unclear how changing geochemical zones and sediment depth influence their community structure. We mapped the community composition and abundance of SRMs by amplicon sequencing and quantifying the dsrB gene, which encodes dissimilatory sulfite reductase subunit beta, in sediment samples covering different vertical geochemical zones ranging from the surface sediment to the deep sulfate-depleted subsurface at four locations in Aarhus Bay, Denmark. SRMs were present in all geochemical zones, including sulfate-depleted methanogenic sediment. The biggest shift in SRM community composition and abundance occurred across the transition from bioturbated surface sediments to nonbioturbated sediments below, where redox fluctuations and the input of fresh organic matter due to macrofaunal activity are absent. SRM abundance correlated with sulfate reduction rates determined for the same sediments. Sulfate availability showed a weaker correlation with SRM abundances and no significant correlation with the composition of the SRM community. The overall SRM species diversity decreased with depth, yet we identified a subset of highly abundant community members that persists across all vertical geochemical zones of all stations. We conclude that subsurface SRM communities assemble by the persistence of members of the surface community and that the transition from the bioturbated surface sediment to the unmixed sediment below is a main site of assembly of the subsurface SRM community.IMPORTANCE Sulfate-reducing microorganisms (SRMs) are key players in the marine carbon and sulfur cycles, especially in coastal sediments, yet little is understood about the environmental factors controlling their depth distribution. Our results suggest that macrofaunal activity is a key driver of SRM abundance and community structure in marine sediments and that a small subset of SRM species of high relative abundance in the subsurface SRM community persists from the sulfate-rich surface sediment to sulfate-depleted methanogenic subsurface sediment. More generally, we conclude that SRM communities inhabiting the subsurface seabed assemble by the selective survival of members of the surface community.


Is there a role for hyperbaric oxygen therapy in the treatment of refractory wounds of rare etiology?
Delayed wound healing indicates wounds that have failed to respond to more than 4-6 weeks of comprehensive wound care. Wounds with delayed healing are a major source of morbidity and a major cost to hospital and community healthcare providers. Hyperbaric oxygen therapy (HBOT) is a treatment designed to increase the supply of oxygen to wounds and has been applied to a variety of wound types. This article reviews the place of HBOT in the treatment of non-healing vasculitic, calcific uremic arteriolopathy (CUA), livedoid vasculopathy (LV), pyoderma gangrenosum (PG) ulcers. We searched electronic databases for research and review studies focused on HBOT for the treatment of delayed healing ulcers with rare etiologies. We excluded HBOT for ulcers reviewed elsewhere. We included a total of three case series and four case reports including 63 participants. Most were related to severe, non-healing ulcers in patients with vasculitis, CUA, LV, and PG. There was some evidence that HBOT may improve the healing rate of wounds by increasing nitric oxide (NO) levels and the number of endothelial progenitor cells in the wounds. HBOT may also improve pain in these ulcers. We recommend the establishment of comprehensive and detailed wound care registries to rapidly collect prospective data on the use of HBOT for these problem wounds. There is a strong case for appropriately powered, multi-centre randomized trials to establish the true efficacy and cost-effectiveness of HBOT especially for vasculitis ulcers that have not improved following immunosuppressive therapy.


Development of a perfusion culture system for production of biologicals using contact dependent cells.
Design and evaluation of a new type of culture vessel for contact dependent cells is described. Cells are grown on both sides of stationary glass plates in a laminar flow of medium supplied from separate reservoirs. Construction and operation of vessels ranging in growth surface from 0.02 to 10 m2 is relatively simple and there are no moving parts. Cells can be inspected by low power microscopy in the usual way. Evaluation of this equipment was carried out using the human diploid cell line MRC-5. Saturation densities of up to 1 million cells per square centimetre of growth surface were obtained where appropriate medium volumes were used. Application of this sytem to other cell types and the production of biological products is briefly discussed.


Discontinuation of Pneumocystis jirovecii pneumonia prophylaxis with CD4 count <200 cells/µL and virologic suppression: a systematic review.
HIV viral load (VL) is currently not part of the criteria for Pneumocystis jirovecii pneumonia (PCP) prophylaxis discontinuation, but suppression of plasma viremia with antiretroviral therapy may allow for discontinuation of PCP prophylaxis even with CD4 count <200 cells/µL. A systematic review was performed to determine the incidence of PCP in HIV-infected individuals with CD4 count <200 cells/µL and fully suppressed VL on antiretroviral therapy but not receiving PCP prophylaxis. Four articles examined individuals who discontinued PCP prophylaxis with CD4 count <200 cells/µL in the context of fully suppressed VL on antiretroviral therapy. The overall incidence of PCP was 0.48 cases per 100 person-years (PY) (95% confidence interval (CI) (0.06-0.89). This was lower than the incidence of PCP in untreated HIV infection (5.30 cases/100 PY, 95% CI 4.1-6.8) and lower than the incidence in persons with CD4 count <200 cells/µL, before the availability of highly active antiretroviral therapy (HAART), who continued prophylaxis (4.85/100 PY, 95% CI 0.92-8.78). In one study in which individuals were stratified according to CD4 count <200 cells/µL, there was a greater risk of PCP with CD4 count ≤100 cells/µL compared to 101-200 cells/µL. Primary PCP prophylaxis may be safely discontinued in HIV-infected individuals with CD4 count between 101-200 cells/µL provided the VL is fully suppressed on antiretroviral therapy. However, there are inadequate data available to make this recommendation when the CD4 count is ≤100 cells/µL. A revision of guidelines on primary PCP prophylaxis to include consideration of the VL is merited.


PKA-dependent regulation of mKv1.1, a mouse Shaker-like potassium channel gene, when stably expressed in CHO cells.
Potassium (K) channels are important regulators of cellular physiology and can themselves be modulated by phosphorylation. We have investigated the potential protein kinase A (PKA) regulation of mKv1.1, a mouse Shaker-like K channel gene, when it is expressed in stably transfected Chinese hamster ovary (CHO) cell lines. Whole-cell patch-clamp records show that expression of mKv1.1 gives rise to a rapidly activating, sustained K+ current, referred to classically as a delayed rectifier-type current. In order to study the effects of PKA, we compared cell lines transfected with mKv1.1 alone with lines cotransfected with both mKv1.1 and a plasmid encoding a dominant negative mutation in the regulatory subunit of PKA. These mutant regulatory subunits bind to endogenous catalytic subunits of PKA but do not respond to cAMP, thereby causing a chronic reduction in the basal PKA activity in these cells. We found that mKv1.1 current kinetics are unaltered but current density is 3.4-fold higher in the cell lines expressing mutant regulatory subunit than in lines expressing only mKv1.1. RNase protection assays indicate that levels of the specific RNA for mKv1.1 are increased almost twofold in the lines expressing mutant regulatory subunit over the lines expressing mKv1.1 only. Further, the levels of mKv1.1 protein, assayed using an mKv1.1 channel-specific antibody, are increased by almost a factor of 3 between the two types of cell lines. These results suggest that PKA can regulate mKv1.1 channel expression by changing steady-state levels of RNA and by other posttranscriptional mechanisms.


Steroid characteristics of mineralocorticoid adrenocortical hypertension.
Adrenocortical causes of hypertension are established by examining the mineralocorticoid hormones produced in the zona glomerulosa and zona fasciculata. In the zona glomerulosa, aldosterone excess leads to hypertension, hypokalemia, and suppressed plasma renin activity, with increased concentrations of urinary aldosterone (either as the 18-glucuronide or free aldosterone) as an index of its production. Identifying a tumor by computed tomography scan verifies the diagnosis of a correctable lesion. If no tumor is found, several maneuvers are used to identify primary adrenal hyperplasia, a disorder with autonomous aldosterone production, for which reduction of adrenal mass is curative. The zona fasciculata has two major pathways: the 17-deoxy pathway, where deoxycorticosterone (DOC) and corticosterone are the significant steroids, and the 17-hydroxy pathway, which leads to cortisol production. Tumors of the 17-deoxy pathway, DOC-producing adenomas, have increased concentrations of DOC and its precursor steroids, normal concentrations of cortisol, and suppression of aldosterone production secondary to suppression of the renin system. Two enzymatic defects in the zona fasciculata, 11 beta- and 17 alpha-hydroxylase deficiency, can be first readily identified by the virilization in the former, hypogonadal features in the latter. Steroid patterns are diagnostic. DOC is produced in excess in both deficiencies and is the cause of the hypertension. Deficient or impaired 11 beta-hydroxy steroid dehydrogenase in the apparent mineralocorticoid excess syndrome or after licorice ingestion retards the conversion of cortisol to inactive cortisone in the kidney, leading to mineralocorticoid hypertension; this leads to suppression of the renin system and subsequently of aldosterone.


Circulating vitamin D and colorectal adenomas in Japanese men.
Accumulating evidence suggests that vitamin D has anticarcinogenic effects. However, it is unclear whether the nutrient is involved in the early stage of colorectal carcinogenesis. We examined the association between circulating vitamin D concentrations and colorectal adenomas in Japanese men. The study subjects comprised 656 cases of colorectal adenomas and 648 controls with normal colonoscopy among male self defense officials receiving a pre-retirement health examination between 1997 and 2004. Plasma or serum levels of 25-hydroxyvitamin D [25(OH)D] were measured using a radioimmunoassay method. Logistic regression analysis was used to obtain odds ratios (OR) and 95% confidence intervals (CI) with adjustment for potential confounding variables. Overall, there was no measurable association between circulating 25(OH)D concentrations and colorectal adenomas. When the analysis was restricted to subjects whose blood was taken during the winter season (November-April), the prevalence odds of colorectal adenomas for the highest versus lowest quartile of 25(OH)D was statistically significantly decreased (OR = 0.58; 95% CI = 0.34-0.99). The reduction was more pronounced for the rectum (OR = 0.22) and distal colon (OR = 0.47) than for proximal colon (OR = 0.70). During the summer season (May-October), higher levels of 25(OH)D were associated with an increased odds of small, but not large, adenomas. The present study adds to evidence that high levels of circulating vitamin D measured during darker season is associated with decreased prevalence of adenomas in the distal sites of the colorectum.


[Two cases of fluent aphasia with selective difficulty of syllable identification].
We report two aphasic patients who could discriminate Japanese syllables but could not identify them. Case 1 was a 51-year-old right handed woman with 12-year education. Case 2 was a 50-year-old right handed man with 9-year education. They developed fluent aphasia after a cerebral infarction. Brain MRI of case 1 revealed widely distributed lesions including inferior frontal, superior temporal, angular and supramarginal gyri. Lesions revealed by Brain CT in case 2 included the left superior and middle temporal, angular and supramarginal gyri. Both showed severe impairment of repetition and confrontation naming. No difference of performance was present between repetition of single syllables and polysyllabic words. On the contrary, oral reading of Kana characters were preserved. We examined their ability to perceive syllables in detail. In the discrimination task, they judged whether a pair of heard syllables was same or different. Case 1 was correct in 85% of the tasks and case 2 in 98%. In an identification task, they heard a syllable and chose a corresponding Kana, Kanji, or picture out of 10 respective candidates. Case 1 was correct only in 30% and case 2 in 50% of these tasks. On the other hand, selection of a correct target in response to a polysyllabic word was much better, i.e. 70% in case 1 and 90% in case 2. Based on these data we concluded that (1) syllabic identification is a different process from syllabic discrimination, and (2) comprehension of a polysyllabic word can be achieved even when the precise phonological analysis of continuent syllables are impaired.


[Aspartate, alanine, glutamate, malate glucose-6-phosphate dehydrogenases in Candida albicans].
Multiple molecular forms of aspartate, alanine, glutamate, malate and glucose-6-phosphate dehydrogenases were studied in the course of Candida albicans growth using electrophoresis in polyacrylamide gel. The isozyme spectrum and the activity of dehydrogenases were found to depend on the cultural age and the cofactor being used (NAD or NADP). The protein-antigenic spectrum and the composition of multiple molecular forms of dehydrogenases were compared in C. albicans in the course of its growth in order to characterize more completely the properties of the culture at a certain growth phase.


Xanthones from the botanical dietary supplement mangosteen (Garcinia mangostana) with aromatase inhibitory activity.
Twelve xanthone constituents of the botanical dietary supplement mangosteen (the pericarp of Garcinia mangostana) were screened using a noncellular, enzyme-based microsomal aromatase inhibition assay. Of these compounds, garcinone D (3), garcinone E (5), alpha-mangostin (8), and gamma-mangostin (9) exhibited dose-dependent inhibitory activity. In a follow-up cell-based assay using SK-BR-3 breast cancer cells that express high levels of aromatase, the most potent of these four xanthones was gamma-mangostin (9). Because xanthones may be consumed in substantial amounts from commercially available mangosteen products, the consequences of frequent intake of mangosteen botanical dietary supplements require further investigation to determine their possible role in breast cancer chemoprevention.


Inonotus obliquus attenuates histamine-induced microvascular inflammation.
Cell-to-cell communication is a key element of microvascular blood flow control, including rapidly carrying signals through the vascular endothelium in response to local stimuli. This cell-to-cell communication is negatively impacted during inflammation through the disruption of junctional integrity. Such disruption is associated with promoting the onset of cardiovascular diseases as a result of altered microvascular blood flow regulation. Therefore, understanding the mechanisms how inflammation drives microvascular dysfunction and compounds that mitigate such inflammation and dysfunction are of great interest for development. As such we aimed to investigate extracts of mushrooms as potential novel compounds. Using intravital microscopy, the medicinal mushroom, Inonotus obliquus was observed, to attenuate histamine-induced inflammation conducted vasodilation in second-order arterioles in the gluteus maximus muscle of C57BL/6 mice. Mast cell activation by C48/80 similarly disrupted endothelial junctions and conducted vasodilation but only histamine was blocked by the histamine antagonist, pyrilamine not C48/80 suggesting the importance of mast cell activation. Data presented here supports that histamine induced inflammation is a major disruptor of junctional integrity, and highlights the important anti-inflammatory properties of Inonotus obliquus focusing future assessment of mast cells as putative target for Inonotus obliquus.


Establishment of a hepatocyte-kupffer cell coculture model for assessment of proinflammatory cytokine effects on metabolizing enzymes and drug transporters.
Elevated levels of proinflammatory cytokines associated with infection and inflammation can modulate cytochrome P450 enzymes, leading to potential disease-drug interactions and altered small-molecule drug disposition. We established a human-derived hepatocyte-Kupffer cell (Hep:KC) coculture model to assess the indirect cytokine impact on hepatocytes through stimulation of KC-mediated cytokine release and compared this model with hepatocytes alone. Characterization of Hep:KC cocultures showed an inflammation response after treatment with lipopolysaccharide and interleukin (IL)-6 (indicated by secretion of various cytokines). Additionally, IL-6 exposure upregulated acute-phase proteins (C-reactive protein, alpha-1-acid glycoprotein, and serum amyloid A2) and downregulated CYP3A4. Compared with hepatocytes alone, Hep:KC cocultures showed enhanced IL-1β-mediated effects but less impact from both IL-2 and IL-23. Hep:KC cocultures treated with IL-1β exhibited a higher release of proinflammatory cytokines, an increased upregulation of acute-phase proteins, and a larger extent of metabolic enzyme and transporter suppression. IC50 values for IL-1β-mediated CYP3A4 suppression were lower in Hep:KC cocultures (98.0-144 pg/ml) compared with hepatocytes alone (IC50 > 5000 pg/ml). Cytochrome suppression was preventable by blocking IL-1β interaction with IL-1R1 using an antagonist cytokine or an anti-IL-1β antibody. Unlike IL-1β, IL-6-mediated effects were comparable between hepatocyte monocultures and Hep:KC cocultures. IL-2 and IL-23 caused a negligible inflammation response and a minimal inhibition of CYP3A4. In both hepatocyte monocultures and Hep:KC cocultures, IL-2RB and IL-23R were undetectable, whereas IL-6R and IL-1R1 levels were higher in Hep:KC cocultures. In summary, compared with hepatocyte monocultures, the Hep:KC coculture system is a more robust in vitro model for studying the impact of proinflammatory cytokines on metabolic enzymes.


Bilateral diaphragm paralysis after cardiac surgery with topical hypothermia.
Bilateral diaphragm paralysis is a rare but important complication of open heart surgery. Two cases were found among 360 prospectively studied patients undergoing open heart surgery during one year. Both patients had insulin dependent diabetes with peripheral neuropathy and this may have contributed to their diaphragm paralysis. The patients were studied postoperatively for one year with measurements of lung function, nocturnal oximetry, diaphragmatic function, and phrenic nerve conduction. Treatment with intermittent positive airway pressure ventilation by nasal mask was effective in both patients. After nine months one patient had recovered completely with normal phrenic nerve conduction and diaphragmatic function; the other continues most of his normal daytime activities, but still requires nasal positive airway pressure ventilation for six hours at night.


Neuronal cell types in the thalamic intralaminar central lateral nucleus of the cat.
The existence of Golgi type II neurons was verified in the anterior intralaminar central lateral (CL) nucleus of the cat thalamus, and its projection cell types were identified, by means of Golgi impregnation. CL principal neurons were found to display a large- or medium-sized cell body and a radiate dendritic pattern. Their primary dendrites were limited in number, and had a rather long course; they were poorly ramified. The axons of principal neurons were impregnated only occasionally and for a short distance. Projection neurons of the 'bushy' or tufted type, described in the main thalamic sensory nuclei, were not identified in the CL in the present study. Typical Golgi type II neurons were found throughout CL. They were mainly small-sized, and displayed a rich dendritic arborization characterized by dendritic appendages. The axons of Golgi type II neurons were seen to give rise to extensive local arborizations. The present findings indicate that in the cat CL, principal cells are mainly represented by radiate neurons. Typical local circuit neurons also are evident in CL, suggesting that the activity of anterior intralaminar structures is regulated by intrinsic mechanisms similar to those operating in the main thalamic relay nuclei.


[Effect an ACTH analog on the processes of learning and memory in rats].
On the basis of the idea of the important role of neurotransmitter systems in realization of neuropeptide effects, the participation was studied of the monoaminergic systems in the mechanisms of the ACTH analogue influence on the processes of learning and memory in control animals and animals with a changed functional state of the monoaminergic systems. In parallel the influence was studied of the ACTH analogue on the content of the endogenic monoamines in various brain structures of rats. It has been shown that administration of the ACTH analogue in a dose of 10 mcg affects the elaboration and preservation of conditioned reflexes (CRs) of passive avoidance, CRs of two-side avoidance and labyrinth CRs only in conditions of changed functional state of the monoaminergic systems. Amnesia, usually elicited by 5-oxytryptophane and disulfiram is prevented by administration of the ACTH analogue. Administration of the ACTH analogue is accompanied by the intensification of serotonine metabolism in the midbrain and medulla and by an increase of noradrenaline content in the hypothlamus.


Which are the leukaemic cells?
Two features are known about acute myeloid leukaemia in man: (1) the long time-scale from identifiable leukaemogenic stimulus and onset of the clinical disease and (2) the successful induction and duration of long clinical remission. These indicate three probabilities: First, that the target cell for leukaemogenetic insult (in AML) is the pluripotent stem cell; second, that the leukaemic stem line is a small minority population within the total leukaemic cell mass; third, when the leukaemic stem line is not greatly exceeding the normal stem cell numbers, its proliferation may be still under partial control.


[Decreased ATP-synthesis ability of brain mitochondria in spontaneously hypertensive rats].
It has been shown previously that a decrease of ATP amount and changed balance of other macroergic phosphates occurs in different tissues of spontaneously hypertensive rats (SHR) compared with control normotensive rats (WKY). To assess the ability of SHR brain isolated mitochondria to synthesize ATP and to elucidate its relation to extramitochondrial calcium concentration. The present work shows for the first time that SHR brain mitochondria initially differ from WKY ones by decreased (by 30%) ATP synthesis rate. When calcium concentration is increased up to 20 or 40 micromole/l ATP synthesis rates in WKY and SHR mitochondria are about one half compared with the condition when the medium does not contain calcium, but the differences between SHR and WKY remain the same. Decreased ATP synthesis rate in SHR mitochondria may be considered as the main cause of cell energy deficiency observed in experimental variant of primary arterial hypertension.


Pulmonary function changes after interscalene brachial plexus anesthesia with 0.5% and 0.75% ropivacaine: a double-blinded comparison with 2% mepivacaine.
The purpose of this investigation was to compare, in a prospective, double-blinded fashion, 0.5% and 0.75% ropivacaine with 2% mepivacaine to determine their effects on respiratory function during interscalene brachial plexus (IBP) anesthesia. With ethical committee approval and written, informed consent, 30 healthy patients presenting for elective shoulder capsuloplastic or acromioplastic procedures were randomized to receive IBP anesthesia by 20 mL of either 0.5% ropivacaine (n = 10), 0.75% ropivacaine (n = 10), or 2% mepivacaine (n = 10). Block onset time, pulmonary function variables, ipsilateral hemidiaphragmatic motion (ultrasonographic evaluation), and first requirement of postoperative analgesic were evaluated. Surgical anesthesia (loss of pinprick sensation from C4 to C7 and motor block of the shoulder joint) was achieved later with 0.5% ropivacaine than with either 0.75% ropivacaine or 2% mepivacaine (P < 0.05), whereas the first pain medication was requested later with both ropivacaine concentrations than with mepivacaine (P < 0.0005). No differences in quality of the block or patient acceptance were observed in the three groups. All 30 patients had ipsilateral hemidiaphragmatic paresis and large mean decreases in forced vital capacity (ropivacaine 0.5%: 40% +/- 17%, ropivacaine 0.75%: 41% +/- 22%, mepivacaine 2%: 39% +/- 21%) and forced expiratory volume at 1 s (ropivacaine 0.5%: 30% +/- 19%, ropivacaine 0.75%: 38% +/- 26%, mepivacaine 2%: 40% +/- 10%). We conclude that, when performing IBP anesthesia, 0.5% ropivacaine does not decrease the incidence of ipsilateral paresis of the hemidiaphragm compared with 0.75% ropivacaine and 2% mepivacaine. During the first 30 min after placing interscalene brachial plexus anesthesia, 0.5% ropivacaine does not provide clinically relevant advantages in terms of pulmonary function changes compared with either 0.75% ropivacaine or 2% mepivacaine. However, 0.75% ropivacaine allows a short onset, similar to that of mepivacaine, with long postoperative analgesia.


Recent co-evolution of two pandemic plant diseases in a multi-hybrid swarm.
Most plant pathogens exhibit host specificity but when former barriers to infection break down, new diseases can rapidly emerge. For a number of fungal diseases, there is increasing evidence that hybridization plays a major role in driving host jumps. However, the relative contributions of existing variation versus new mutations in adapting to new host(s) is unclear. Here we reconstruct the evolutionary history of two recently emerged populations of the fungus Pyricularia oryzae that are responsible for two new plant diseases: wheat blast and grey leaf spot of ryegrasses. We provide evidence that wheat blast/grey leaf spot evolved through two distinct mating episodes: the first occurred ~60 years ago, when a fungal individual adapted to Eleusine mated with another individual from Urochloa. Then, about 10 years later, a single progeny from this cross underwent a series of matings with a small number of individuals from three additional host-specialized populations. These matings introduced non-functional alleles of two key host-specificity factors, whose recombination in a multi-hybrid swarm probably facilitated the host jump. We show that very few mutations have arisen since the founding event and a majority are private to individual isolates. Thus, adaptation to the wheat or Lolium hosts appears to have been instantaneous, and driven entirely by selection on repartitioned standing variation, with no obvious role for newly formed mutations.


Overexpression of integrin-β1 in leiomyoma promotes cell spreading and proliferation.
Uterine leiomyoma, the most common tumors found in the women of the reproductive age, may cause abnormal uterine bleeding and be life threatening. Compared with myometrium, leiomyoma contains excessive extracellular matrix (ECM). However, the pathological roles of ECM in the development of leiomyoma remain largely unknown. Integrins are the major adhesion molecules on cell surface to interact with ECM. The interactions of ECM with integrins regulate cell adhesion and initiate signals for cell growth, differentiation, and migration. The aim of this study was to investigate the expression and functional role of integrin-β1 in leiomyoma pathogenesis. Levels of integrin-β1 protein were determined by Western blotting in paired normal and leiomyomal tissues (n = 15). Knockdown of integrin-β1 and inhibition of ECM-integrin interaction by disintegrin were used to evaluate the impact of integrin-β1 in cell adhesion, spreading, and proliferation. Levels of integrin-β1 were significantly up-regulated in leiomyomal cells compared with their normal counterparts. Knockdown of integrin-β1 did not affect cell adhesion on fibronectin or laminin matrix but significantly inhibits cell spreading ability. Consistent with this notion, the phosphorylation of focal adhesion kinase and the recruitment of paxillin to the focal contact were decreased in integrin-β1 knockdown cells, which attenuates contraction force. The inability of cell spreading leads to inhibition of cyclin D1 expression and impedes cell cycle progression. More importantly, disruption of ECM-integrin interaction by the small protein, disintegrin inhibited cyclin D1 expression and cell proliferation. These data demonstrate that integrin-β1 is a critical ligand to enhance cell-ECM contact force and thus promotes cell proliferation. Disruption of ECM-integrin-β1 signaling may serve as an option to inhibit the progression of leiomyoma.


Clinical, ultrasonographic, and laboratory findings in 12 llamas and 12 alpacas with malignant round cell tumors.
Clinical signs, duration of illness, clinicopathologic findings, and ultrasonographic findings were evaluated in 12 llamas and 12 alpacas with malignant round cell tumors (MRCT). All but 1 animal died or was euthanized. Common clinical findings were anorexia, recumbency or weakness, and weight loss or poor growth. Peripheral lymphadenomegaly occurred in only 7 animals and was detected more often at necropsy than during physical examination. Common clinicopathologic abnormalities were hypoalbuminemia, acidosis, azotemia, anemia, hyperglycemia, and neutrophilia. Ultrasonography detected tumors in 4/6 animals. Cytologic evaluation of fluid or tissue aspirates or histopathology of biopsy tissue was diagnostic in 5/6 cases. A clinical course of 2 wk or less prior to death or euthanasia was more common in animals ≤ 2 y of age (9/11) than in older animals (6/13). Regular examination of camelids to include clinical pathology and evaluation of peripheral lymph nodes may result in early detection of MCRT.


Mechanism and Progress of Natural Products in the Treatment of NAFLD-Related Fibrosis.
Nonalcoholic fatty liver disease (NAFLD) has emerged as the most prevalent chronic liver disorder worldwide, with liver fibrosis (LF) serving as a pivotal juncture in NAFLD progression. Natural products have demonstrated substantial antifibrotic properties, ushering in novel avenues for NAFLD treatment. This study provides a comprehensive review of the potential of natural products as antifibrotic agents, including flavonoids, polyphenol compounds, and terpenoids, with specific emphasis on the role of Baicalin in NAFLD-associated fibrosis. Mechanistically, these natural products have exhibited the capacity to target a multitude of signaling pathways, including Hedgehog, Wnt/β-catenin, TGF-β1, and NF-κB. Moreover, they can augment the activities of antioxidant enzymes, inhibit pro-fibrotic factors, and diminish fibrosis markers. In conclusion, this review underscores the considerable potential of natural products in addressing NAFLD-related liver fibrosis through multifaceted mechanisms. Nonetheless, it underscores the imperative need for further clinical investigation to authenticate their effectiveness, offering invaluable insights for future therapeutic advancements in this domain.


Thromboprophylaxis in hip fracture surgery: a pilot study comparing danaparoid, enoxaparin and dalteparin. The TIFDED Study Group.
A pilot study was performed to compare the thromboprophylactic effect of danaparoid, enoxaparin and dalteparin in patients with hip fracture. The study was a prospective, randomised assessor-blind, four-centre trial. Prophylaxis was given for 9-11 days, whereafter bilateral phlebography was performed. A total of 197 patients were randomised. There were no statistically significant differences in the frequency of deep vein thrombosis, in blood loss or bleeding complications between the three prophylaxis groups. In conclusion, this moderately sized study revealed no statistically significant difference in efficacy or safety between danaparoid, enoxaparin and dalteparin in patients undergoing hip fracture surgery.


Orthostatic hypotension, orthostatic intolerance and frailty: The Irish Longitudinal Study on Aging-TILDA.
Because frailty may represent impaired response to physiological stress we explored the associations between frailty and orthostatic hypotension (OH), and orthostatic intolerance (OI). This study was based on a cross-sectional analysis of 5692 community dwelling adults aged 50 years and older included in wave 1 of the Irish Longitudinal Study on Aging. Frailty was assessed using both the phenotypic (FP) and frailty index (FI) models. OH was defined as a drop of ≥20 mmHg in systolic blood pressure or a drop of ≥10 mmHg diastolic pressure on standing from a seated position. OI was defined as reporting feeling dizzy, light headed or unsteady during this test. 346 (6.1%) participants had OH and 381 (6.7%) participants had OI. The prevalence OH in frail participants was 8.9%, compared to 5% in robust. Similarly the prevalence of OI was 14.3% in frail and 5.7% in robust participants. After adjustment for age and gender, OH was not significantly related to the FP (OR=1.10 95% CI=0.67, 1.81). Conversely OI was (OR=1.80 95% CI=1.13, 2.87), even after adjustment for age, gender, cardiovascular factors and mental health. In fully adjusted models OI remained related to slowness and low muscle strength and to higher FI scores. These data suggest OI symptoms in older adults may reflect various important underlying health deficits, indicative of increasing levels of frailty. Further assessment of frailty in patients experiencing OI is a potential opportunity for early intervention to delay functional decline.


[Tactics and indications for surgical treatment of patients with combined extra- and intracranial lesions of the carotid arteries].
The authors present the results of surgical treatment of patients with concomitant extra-intracranial and bilateral lesions of the carotid arteries. The patients underwent 163 operations for establishment of extra-intracranial microanastomosis (EICMA) including reconstruction of the extracranial segments with EICMA establishment (75 cases). In 66 cases, EICMA was performed for contralateral stenosis of the internal carotid artery (ICA). Of these, 22 patients later underwent carotid endarterectomy. In 14 patients, EICMA was established as the first stage before reconstruction of the iliofemoral segment. The operation was also performed in 8 patients with occlusion of the median cerebral artery or with the crimp of the carotid arteries. Examinations that included ultrasound dopplerography, transcranial dopplerography, studies of the cerebral blood flow according to 133Xe clearance and angiography were carried out. A study was made of the long-term results of the surgical treatment (from 0.5 to 5 years). It has been discovered that to estimate indications for surgical treatment, it is necessary that the data on the regional blood flow (RBF) be taken into account. In patients with stable cerebral hemodynamics according to the RBF that characterizes an uneventful course of ICA occlusion and in the decompensated type most typical of patients with pronounced postinsult neurological dificit, the establishment of EICMA is not indicated. Surgical treatment is most desirable in ICA occlusion (with preliminary reconstruction of the extracranial segments provided they are impaired) and in patients with unstable cerebral hemodynamics most typical of patients with transitory ischemic attacks, dyscirculatory encephalopathy, and minor brain stroke with chiefly mild residual phenomena of ischemic brain stroke.


5. What do we do now? Implications for the clinical practice.
The primary objective of this supplement is to provide eye care practitioners (ECPs) with the latest research and experience on topics related to contact lens use and patient eye health. This section examines a variety of topics, including the challenges of keeping abreast of the scientific literature, recognizing data that are borne from well-designed studies, and keys to their implementation in clinical practice. Insights are also provided on how eye care is practiced in Asia, where regulations and patient perceptions result in care that is delivered much differently than in North America and Europe. The role of silicone hydrogel (SiHy) lenses in the clinical practice and how advances with this lens material may shape future lens prescribing are then reviewed. The final part examines the current thinking regarding corneal infiltrates, microbial keratitis (MK), and multipurpose solution (MPS)-related corneal staining, and how ECPs should approach these issues.


Omega-3 fatty acids and selenium as coronary heart disease risk modifying factors in Asian Indian and Chinese males.
Asian Indian men are reported to have a higher incidence of coronary heart disease than men of other ethnic groups worldwide. Among the many hypotheses, one possible risk factor may be related to their dietary habits. This study estimated the plasma concentrations of fatty acids, antioxidant vitamins, and selenium in Indians and Chinese of Singapore. The study population consisted of 145 Indian men and 147 Chinese men ages 26 to 79 y from a cross-sectional survey, the National University of Singapore Heart Study. Our findings indicated that Indians had lower plasma concentrations of docosahexanoic acid (3.07% versus 3.54%, P < 0.001), alpha-linolenic acid (0.48% versus 0.57%, P < 0.001), and total omega-3 fatty acids (4.71% versus 5.27%, P < 0.001) than did the Chinese. Arachidonic acid was higher in Indians (4.83%) than in the Chinese (4.51%, P = 0.007). The ratio of omega-3 acid to omega-6 fatty acid was also lower in Indians (0.15) than in the Chinese (0.16, P = 0.007). There were no significant differences in the concentrations of monounsaturated fatty acids, but saturated fatty acids were higher in Indians (39.17%) than in the Chinese (38.28%, P < 0.001). Analysis of vitamins A, C, and E showed no significant differences between Indians (0.67, 5.72, and 13.04 mg/L, respectively) and Chinese (0.68, 6.48, and 12.71 mg/L, respectively); however, serum concentration of selenium in Indians (117.49 microg/L) was significantly lower than in the Chinese (126.72 microg/L, P < 0.001). The results suggest that lower plasma concentrations of omega-3 fatty acids and selenium and higher concentrations of arachidonic acid and saturated fatty acids in Indians may reflect lower intakes of marine foods and, as a consequence, higher susceptibility to coronary heart disease.


A possible interaction between lithium and diltiazem: case report.
The calcium channel blockers are a group of drugs that have recently become available for the treatment of angina. They act by blocking the entry of calcium into cells during depolarization. Lithium has electrochemical qualities similar to calcium and influences calcium metabolism and calcium transport. The case reported here documents the use of diltiazem in a patient treated with lithium. A possible synergistic drug interaction occurring between diltiazem and lithium is reported.


Developing a language screening test for Arabic-speaking children.
This report gives an account of general and specific issues associated with developing an Arabic-language screening test for children aged between 3 and 12 years. Challenges, dilemmas and stages in the developmental process are discussed. The development of the Arabic Language Screening Test: Preschool and School Age illustrates the process. Issues and stages included: (1) arriving at a consensus about scope and purpose; (2) conceptualizing tasks and items; (3) specifying item content and structure; (4) field-testing the screening tool in a circumscribed environment; (5) developing a robust scoring and interpretation system for determining Pass or Fail, and (6) establishing the degree of accuracy in differentiating children with and without language disorders. The product is a screening test of verbal and related nonverbal abilities with parallel components for children of preschool (3-5 years) and elementary school age (6-12 years). Normative data were collected for 750 Arabic-speaking children in Jordan and Palestine, distributed fairly equally between the ages of 3 and 12 years. Normative means increased with age and standard deviations decreased. Interscorer agreement (99%), internal consistency (Cronbach alpha = 0.98) and diagnostic sensitivity (over 90% accuracy) were high. Constraints and limitations in the development of the test are described and discussed from both objective and personal perspectives.


Role of sigma 1 receptor in high fat diet-induced peripheral neuropathy.
The neurobiological mechanisms of obesity-induced peripheral neuropathy are poorly understood. We evaluated the role of Sigma-1 receptor (Sig-1R) and NMDA receptor (NMDARs) in the spinal cord in peripheral neuropathy using an animal model of high fat diet-induced diabetes. We examined the expression of Sig-1R and NMDAR subunits GluN2A and GluN2B along with postsynaptic density protein 95 (PSD-95) in the spinal cord after 24-week HFD treatment in both wild-type and Sig-1R-/- mice. Finally, we examined the effects of repeated intrathecal administrations of selective Sig-1R antagonists BD1047 in HFD-fed wild-type mice on peripheral neuropathy. Wild-type mice developed tactile allodynia and thermal hypoalgesia after 24-week HFD treatment. HFD-induced peripheral neuropathy correlated with increased expression of GluN2A and GluN2B subunits of NMDARs, PDS-95, and Sig-1R, as well as increased Sig-1R-NMDAR interaction in the spinal cord. In contrast, Sig-1R-/- mice did not develop thermal hypoalgesia or tactile allodynia after 24-week HFD treatment, and the levels of GluN2A, GluN2B, and PSD-95 were not altered in the spinal cord of HFD-fed Sig-1R-/- mice. Finally, repeated intrathecal administrations of selective Sig-1R antagonists BD1047 in HFD-fed wild-type mice attenuated peripheral neuropathy. Our results suggest that obesity-associated peripheral neuropathy may involve Sig-1R-mediated enhancement of NMDAR expression in the spinal cord.


Construction of a Realistic, Whole-Body, Three-Dimensional Equine Skeletal Model using Computed Tomography Data.
Therapies based upon whole-body biomechanical assessments are successful for injury prevention and rehabilitation in human athletes. Similar approaches have rarely been used to study equine athletic injury. Degenerative osteoarthritis caused by mechanical stress can originate from chronic postural dysfunction, which, because the primary dysfunction is often distant from the site of tissue injury, is best identified through modeling whole-body biomechanics. To characterize whole-body equine kinematics, a realistic skeletal model of a horse was created from equine computed tomography (CT) data that can be used for functional anatomical and biomechanical modeling. Equine CT data were reconstructed into individual three-dimensional (3D) data sets (i.e., bones) using 3D visualization software and assembled into a complete 3D skeletal model. The model was then rigged and animated using 3D animation and modeling software. The resulting 3D skeletal model can be used to characterize equine postures associated with degenerative tissue changes as well as to identify postures that reduce mechanical stress at the sites of tissue injury. In addition, when animated into 4D, the model can be used to demonstrate unhealthy and healthy skeletal movements and can be used to develop preventative and rehabilitative individualized therapies for horses with degenerative lamenesses. Although the model will soon be available for download, it is currently in a format that requires access to the 3D animation and modeling software, which has quite a learning curve for new users. This protocol will guide users in (1) developing such a model for any organism of interest and (2) using this specific equine model for their own research questions.


Hypophosphatemia: an evidence-based problem-solving approach to clinical cases.
Hypophosphatemia is defined as a serum phosphate level of less than 2.5 mg/dL (0.8 mmol/L). Hypophosphatemia is caused by inadequate intake, decreased intestinal absorption, excessive urinary excretion, or a shift of phosphate from the extracellular to the intracellular compartments. Renal phosphate wasting can result from genetic or acquired renal disorders. Acquired renal phosphate wasting syndromes can result from vitamin D deficiency hyperparathyroidism, oncogenic osteomalecia, and Fanconi syndrome. Genetic disorders of renal hypophosphatemic disorders generally manifest in infancy and are usually transmitted as an X-linked hypophosphatemic rickets. Symptoms of hypophosphatemia are nonspecific and most patients are asymptomatic. Severe hypophosphatemia may cause skeletal muscle weakness, myocardial dysfunction, rhabdomyolysis, and altered mental status. The diagnostic approach to hypophosphatemia should begin with the measurement of fractional phosphate excretion; if greater than 15% in the presence of hypophosphatemia, the diagnosis of renal phosphate wasting is confirmed. Renal phosphate wasting can be divided into 3 types based upon serum calcium levels: primary hyperparathyroidism (high serum calcium level), secondary hyperparathyroidism (low serum calcium level), and primary renal phosphate wasting (normal serum calcium level). Phosphate supplementations are indicated in patients who are symptomatic or who have a renal tubular defect leading to chronic phosphate wasting. Oral phosphate supplements in combination with calcitriol are the mainstay of treatment. Parenteral phosphate supplementation is generally reserved for patient with life-threatening hypophosphatemia (serum phosphate < 2.0 mg/dL). Intravenous phosphate (0.16 mmol/kg) is administered at a rate of 1 mmol/h to 3 mmol/h until a level of 2 mg/dL is reached.


Lecanemab: More Questions Than Answers!
The approval of lecanemab by the US Food and Drug Administration has been touted as a defining moment in the treatment of Alzheimer's disease. Lecanemab, an anti-amyloid beta monoclonal antibody, is the first Alzheimer's disease drug targeting amyloid beta that has shown statistically significant cognitive benefits in phase III trials. However, there have been many questions raised over the clinical relevance of the otherwise minimal cognitive improvements. Furthermore, its rapid approval has been mired in controversy, in addition to the reports of adverse events such as amyloid-related imaging abnormalities and several deaths of participants in the lecanemab trials. Here, we analyze the evidence supporting lecanemab as an amyloid beta therapy and also discuss the concerns raised about its efficacy and safety.


The Covering Radius and a Discrete Surface Area for Non-Hollow Simplices.
We explore upper bounds on the covering radius of non-hollow lattice polytopes. In particular, we conjecture a general upper bound of d/2 in dimension d, achieved by the "standard terminal simplices" and direct sums of them. We prove this conjecture up to dimension three and show it to be equivalent to the conjecture of González-Merino and Schymura (Discrete Comput. Geom. 58(3), 663-685 (2017)) that the d-th covering minimum of the standard terminal n-simplex equals d/2, for every n ≥ d . We also show that these two conjectures would follow from a discrete analog for lattice simplices of Hadwiger's formula bounding the covering radius of a convex body in terms of the ratio of surface area versus volume. To this end, we introduce a new notion of discrete surface area of non-hollow simplices. We prove our discrete analog in dimension two and give strong evidence for its validity in arbitrary dimension.


Clinical application of proteomics in breast cancer: state of the art and perspectives.
Breast cancer is one of the most frequent and deadly cancers in industrialized countries. The identification of accurate biomarkers that improve the screening, diagnosis, prognostication and prediction of therapeutic response or toxicity, and the identification of novel molecular therapeutic targets are crucial. Today, high-throughput molecular techniques permit investigators to systematically interrogate the genome, transcriptome, and proteome of cancer cells. During the past decade, mRNA expression profiling has been successfully applied to the molecular characterization of breast cancers. Application of proteomics-based techniques is also considered crucial for detecting new biomarkers. In this review, we present the proteomics-based methods that have been applied to date to breast cancer samples for diagnostic and prognostic purposes. Despite their current limits, these pioneering techniques are promising. The most important results as well as the current limitations and perspectives are summarized and discussed.


Timing and visual feedback constraints on repetitive finger force production.
While much is known about sequential effects in motor timing, less is understood about whether movement parameters such as force show sequential dependencies. In this study, we examined the effect of timing constraints on repetitive unimanual force production sequences. Ten healthy participants produced a series of pinch grip forces in time to a metronome and to visually specified force amplitudes. Either visual feedback of force produced or the auditory metronome removed 10 s into the experimental trial, with participants performing continued responses for the remaining 20 s. In the continuation trials, a negative lag-1 autocorrelation in the inter-response intervals (IRIs) was observed as is commonly seen in motor timing tasks. However, removal of visual feedback resulted in a systematic increase in mean force output through the course of the trial, resulting in positive lag-1 autocorrelation values. An interaction was found between mean IRI and peak force (PF) magnitude, with greater force variability seen for the larger intervals. However, the imposition of dual force and timing constraints had no effect either on the underlying variability of the PF or on the IRIs. The results are discussed in the context of force and time being independently specified components of a generalized motor program.


Hearing loss caused by progressive degeneration of cochlear hair cells in mice deficient for the Barhl1 homeobox gene.
The cochlea of the mammalian inner ear contains three rows of outer hair cells and a single row of inner hair cells. These hair cell receptors reside in the organ of Corti and function to transduce mechanical stimuli into electrical signals that mediate hearing. To date, the molecular mechanisms underlying the maintenance of these delicate sensory hair cells are unknown. We report that targeted disruption of Barhl1, a mouse homolog of the Drosophila BarH homeobox genes, results in severe to profound hearing loss, providing a unique model for the study of age-related human deafness disorders. Barhl1 is expressed in all sensory hair cells during inner ear development, 2 days after the onset of hair cell generation. Loss of Barhl1 function in mice results in age-related progressive degeneration of both outer and inner hair cells in the organ of Corti, following two reciprocal longitudinal gradients. Our data together indicate an essential role for Barhl1 in the long-term maintenance of cochlear hair cells, but not in the determination or differentiation of these cells.


Management of the regression of papilledema with regional axon loss in idiopathic intracranial hypertension patients.
To use optic coherence tomography (OCT) to evaluate idiopathic intracranial hypertension (IIH) patients with subclinical segmental optic atrophy despite being under apparently effective treatment. IIH patients underwent an OCT examination including the peripapillary retina never fiber layer (RNFL) thickness, ganglion cell complex (GCC) thickness, focal loss volume (FLV) and global loss volume (GLV) of the GCC, and total macular thickness measurements at presentation and at 3, 6, 9, and 12 months after the diagnosis. The obtained data were compared with healthy subjects. Subjects with and without subclinical segmental atrophy at the 12th month were compared according to the demographics, clinical findings, and the OCT parameters recorded at the beginning of the disease. Both eyes of 56 patients with papilledema due to IIH and 50 age- and sex-matched control subjects were included in this prospective case-control study. Regression of papilledema with regional axon loss on the peripapillary RNFL thickness map was found in 37 (33%) eyes in the IIH group. IIH patients with segmental atrophy had the following characteristics when compared to those without segmental atrophy at the beginning of the disease: higher CSF opening pressure, higher grade of papilledema, thicker mean peripapillary RNFL thickness, thinner GCC layer, greater FLV and GLV loss, and severe visual field loss. Axonal loss occurred in the patients despite apparent treatment. It would be appropriate to follow-up with aggressive medical treatment those patients who present with the following characteristics: higher CSF opening pressure, higher grade of papilledema, thicker mean peripapillary RNFL thickness, and thinner GCC.


Intracellular free calcium concentration/force relationship in rabbit inferior vena cava activated by norepinephrine and high K+.
The changes in isometric force and the underlying fluctuations in intracellular free calcium concentration ([Ca2+]i) were monitored simultaneously in thin sheets of rabbit inferior vena cava loaded with the fluorescent Ca2+ indicator fura-2. In resting tissues bathed in physiological saline solution, the estimated [Ca2+]i was approximately 105 nM. The alpha-adrenergic agonist norepinephrine (10 microM) caused an initial rise in [Ca2+]i to 264 nM during force development, which dropped to 216 nM during force maintenance. The maintained norepinephrine-induced increase in force and [Ca2+]i was reversed in Ca2(+)-free (2 mM EGTA) solution. Membrane depolarization by high K+ (80 mM) significantly increased [Ca2+]i to 234 nM. Compared to norepinephrine, high K+ caused about the same steady-state increase in [Ca2+]i, but a smaller increase in force. [Ca2+]i/force curves were constructed at different concentrations of extracellular Ca2+, with either norepinephrine or high K+ as a stimulant. The curve generated with norepinephrine was located to the left of that generated with high K+.


Genetic diversity and population structure assessed by SSR and SNP markers in a large germplasm collection of grape.
The economic importance of grapevine has driven significant efforts in genomics to accelerate the exploitation of Vitis resources for development of new cultivars. However, although a large number of clonally propagated accessions are maintained in grape germplasm collections worldwide, their use for crop improvement is limited by the scarcity of information on genetic diversity, population structure and proper phenotypic assessment. The identification of representative and manageable subset of accessions would facilitate access to the diversity available in large collections. A genome-wide germplasm characterization using molecular markers can offer reliable tools for adjusting the quality and representativeness of such core samples. We investigated patterns of molecular diversity at 22 common microsatellite loci and 384 single nucleotide polymorphisms (SNPs) in 2273 accessions of domesticated grapevine V. vinifera ssp. sativa, its wild relative V. vinifera ssp. sylvestris, interspecific hybrid cultivars and rootstocks. Despite the large number of putative duplicates and extensive clonal relationships among the accessions, we observed high level of genetic variation. In the total germplasm collection the average genetic diversity, as quantified by the expected heterozygosity, was higher for SSR loci (0.81) than for SNPs (0.34). The analysis of the genetic structure in the grape germplasm collection revealed several levels of stratification. The primary division was between accessions of V. vinifera and non-vinifera, followed by the distinction between wild and domesticated grapevine. Intra-specific subgroups were detected within cultivated grapevine representing different eco-geographic groups. The comparison of a phenological core collection and genetic core collections showed that the latter retained more genetic diversity, while maintaining a similar phenotypic variability. The comprehensive molecular characterization of our grape germplasm collection contributes to the knowledge about levels and distribution of genetic diversity in the existing resources of Vitis and provides insights into genetic subdivision within the European germplasm. Genotypic and phenotypic information compared in this study may efficiently guide further exploration of this diversity for facilitating its practical use.


Enzymatically degradable thermogelling poly(alanine-co-leucine)-poloxamer-poly(alanine-co-leucine).
In the search for an enzymatically degradable thermogelling system, we are reporting poly(alanine-co-leucine)-poloxamer-poly(alanine-co-leucine) (PAL-PLX-PAL) aqueous solution. As the temperature increased, the polymer aqueous solution underwent sol-to-gel transition at 20-40 °C in a polymer concentration range of 3.0-10.0 wt %. The amphiphilic polymers of PAL-PLX-PAL form micelles in water, where the hydrophobic PALs form a core and the hydrophilic PLXs form a shell of the micelle. FTIR, circular dichroism, and (13)C NMR spectra suggest that the α-helical secondary structure of PAL is preserved; however, the molecular motion of the PLX significantly decreases in the sol-to-gel transition range of 20-50 °C. The polymer was degraded by proteolytic enzymes such as matrix metalloproteinase and elastase, whereas it was quite stable against cathepsin B, cathepsin C, and chymotrypsin or in phosphate-buffered saline (control). The in situ formed gel in the subcutaneous layer of rats showed a duration of ∼ 47 days, and H&E staining study suggests the histocompatibility of the gel in vivo with a marginal inflammation response of capsule formation. A model drug of bovine serum albumin was released over 1 month by the preset-gel injection method. The thermogelling PAL-PLX-PAL can be a promising biocompatible material for minimally invasive injectable drug delivery.


13C ENDOR Characterization of the Central Carbon within the Nitrogenase Catalytic Cofactor Indicates That the CFe6 Core Is a Stabilizing "Heart of Steel".
Substrates and inhibitors of Mo-dependent nitrogenase bind and react at Fe ions of the active-site FeMo-cofactor [7Fe-9S-C-Mo-homocitrate] contained within the MoFe protein α-subunit. The cofactor contains a CFe6 core, a carbon centered within a trigonal prism of six Fe, whose role in catalysis is unknown. Targeted 13C labeling of the carbon enables electron-nuclear double resonance (ENDOR) spectroscopy to sensitively monitor the electronic properties of the Fe-C bonds and the spin-coupling scheme adopted by the FeMo-cofactor metal ions. This report compares 13CFe6 ENDOR measurements for (i) the wild-type protein resting state (E0; α-Val70) to those of (ii) α-Ile70, (iii) α-Ala70-substituted proteins; (iv) crystallographically characterized CO-inhibited "hi-CO" state; (v) E4(4H) Janus intermediate, activated for N2 binding/reduction by accumulation of 4[e-/H+]; (vi) E4(2H)* state containing a doubly reduced FeMo-cofactor without Fe-bound substrates; and (vii) propargyl alcohol reduction intermediate having allyl alcohol bound as a ferracycle to FeMo-cofactor Fe6. All states examined, both S = 1/2 and 3/2 exhibited near-zero 13C isotropic hyperfine coupling constants, Ca = [-1.3 ↔ +2.7] MHz. Density functional theory computations and natural bond orbital analysis of the Fe-C bonds show that this occurs because a (3 spin-up/3 spin-down) spin-exchange configuration of CFe6 Fe-ion spins produces cancellation of large spin-transfers to carbon in each Fe-C bond. Previous X-ray diffraction and DFT both indicate that trigonal-prismatic geometry around carbon is maintained with high precision in all these states. The persistent structure and Fe-C bonding of the CFe6 core indicate that it does not provide a functionally dynamic (hemilabile) "beating heart"─instead it acts as "a heart of steel", stabilizing the structure of the FeMo-cofactor-active site during nitrogenase catalysis.


Duloxetine is effective in treating depression in multiple sclerosis patients: an open-label multicenter study.
Duloxetine hydrochloride, a dual reuptake inhibitor of serotonin and norepinephrine, was evaluated for its therapeutic efficacy, safety, and tolerability in the treatment of depression in patients with multiple sclerosis (MS). Lifetime depression prevalence approaches 50% in MS patients. The aim of the study was to assess the safety and efficacy of duloxetine for treatment of depression in MS patients. An open-label study evaluated the efficacy of 12 weeks of duloxetine administration (maximal dose = 60 mg/d) in MS patients with clinical depression. The Beck scale score variation after 4 (T1) and 12 (T2) weeks of treatment was used for the primary outcome measurement, whereas secondary outcome was measured using the Modified Fatigue Impact Scale. Safety was evaluated by recording treatment-related adverse events, monitoring vital signs, and recording frequency and reasons for interruption or discontinuation of treatment. Seventy-five patients were enrolled in the study. Sixty-three patients completed the study by continuing duloxetine treatment for 12 weeks (T2). Twelve subjects dropped out of the study because of adverse effects or noncompliance. Nausea was the most common adverse event reported. A significant reduction in the Beck Depression Inventory and Modified Fatigue Impact Scale scores, after both 4 and 12 weeks of therapy, was observed. The results suggest that duloxetine is well tolerated, safe, and effective in reducing depression and fatigue in MS patients.


Cholic Acid-Peptide Conjugates as Potent Antimicrobials against Interkingdom Polymicrobial Biofilms.
Interkingdom polymicrobial biofilms formed by Gram-positive Staphylococcus aureus and Candida albicans pose serious threats of chronic systemic infections due to the absence of any common therapeutic target for their elimination. Herein, we present the structure-activity relationship (SAR) of membrane-targeting cholic acid-peptide conjugates (CAPs) against Gram-positive bacterial and fungal strains. Structure-activity investigations validated by mechanistic studies revealed that valine-glycine dipeptide-derived CAP 3 was the most effective broad-spectrum antimicrobial against S. aureus and C. albicans CAP 3 was able to degrade the preformed single-species and polymicrobial biofilms formed by S. aureus and C. albicans, and CAP 3-coated materials prevented the formation of biofilms. Murine wound and catheter infection models further confirmed the equally potent bactericidal and fungicidal effect of CAP 3 against bacterial, fungal, and polymicrobial infections. Taken together, these results demonstrate that CAPs, as potential broad-spectrum antimicrobials, can effectively clear the frequently encountered polymicrobial infections and can be fine-tuned further for future applications.