Morphological and functional aspects of the epidermis of the sea lamprey Petromyzon marinus throughout development.
The development of the epidermis of sea lamprey Petromyzon marinus along the whole life cycle was studied using conventional staining techniques and lectin histochemistry. The epidermis undergoes variations in morphology and thickness throughout development. The simple cuboidal epithelium found in the epidermis of prolarvae becomes stratified cubic in the adult by increasing the number of cell layers. The cuticle thickness undergoes a steady increase during the larval period. There are changes in the glycoconjugate composition of the three main cell types of the P. marinus epidermis, mucous, granular and skein cells, which are more pronounced after metamorphosis. The Alcian blue-periodic acid Schiff (AB-PAS) histochemical method shows the presence of both acidic and neutral glycoconjugates in the mucous cells, indicating their secretory function. Moreover, lectin analysis reveals a mucous secretion containing glycoconjugates such as sulphated glycosaminoglycans (N-acetylglucosamine and N-acetylgalactosamine) and N-glycoproteins rich in mannose. Although granular cells are AB-PAS negative, they exhibit a similar glycoconjugate composition to the mucous cells. Moreover, granular cells show sialic acid positivity in larvae but this monosaccharide residue is not detected after metamorphosis. The skein cells, a unique cell of lampreys, are negative to AB-PAS staining but they mostly contain l-fucose and sialic acid residues, which also disappear after metamorphosis. The function of the granular and skein cells is still unknown but the role of their glycoconjugate composition is discussed. In addition, a different cellular origin is suggested for these two types of cells.


Pediatric Brain: No Increased Signal Intensity in the Dentate Nucleus on Unenhanced T1-weighted MR Images after Consecutive Exposure to a Macrocyclic Gadolinium-based Contrast Agent.
Purpose To determine the effect of at least five serial injections of the macrocyclic gadolinium-based contrast agent (GBCA) gadoterate meglumine on the signal intensity (SI) of the dentate nucleus (DN) of the pediatric brain on nonenhanced T1-weighted magnetic resonance (MR) images. Materials and Methods In this retrospective, institutional review board-approved study, 41 pediatric patients (age range, 3-17 years) who were imaged in at least five consecutive 1.5-T MR examinations with the exclusive use of gadoterate meglumine (plus a final additional nonenhanced MR imaging examination) were evaluated. SI ratio differences between the first and last MR examination were calculated for DN-to-pons and DN-to-middle cerebellar peduncle (MCP) ratios in a region-of-interest-based analysis, and one-sample t tests were used to examine if the SI ratio differences differed from 0. Bayes factors were calculated to quantify the strength of evidence for each test. Results Patients underwent a mean of 8.6 ± 3.9 GBCA administrations (mean accumulated dose, 32.07 mmol ± 17.62, with an average of 16.7 weeks ± 7.9 between every administration). Both ratio differences did not differ significantly from 0 (DN-to-pons ratio: -0.0012 ± 0.0101, P = .436; DN-to-MCP ratio: 0.0007 ± 0.0088, P = .604), and one-sided Bayes factors provided substantial evidence against an SI ratio increase (0.10 for DN-to-pons ratio; 0.27 for DN-to-MCP ratio). Conclusion No increase of the SI in the DN was found after a mean of 8.6 serial injections of the macrocyclic GBCA gadoterate meglumine in pediatric patients, confirming previous studies that did not find this effect after serial injections of macrocyclic GBCAs in adults. © RSNA, 2017.


Analysis of rare variant population structure in Europeans explains differential stratification of gene-based tests.
There is substantial interest in the role of rare genetic variants in the etiology of complex human diseases. Several gene-based tests have been developed to simultaneously analyze multiple rare variants for association with phenotypic traits. The tests can largely be partitioned into two classes - 'burden' tests and 'joint' tests - based on how they accumulate evidence of association across sites. We used the empirical joint site frequency spectra of rare, nonsynonymous variation from a large multi-population sequencing study to explore the effect of realistic rare variant population structure on gene-based tests. We observed an important difference between the two test classes: their susceptibility to population stratification. Focusing on European samples, we found that joint tests, which allow variants to have opposite directions of effect, consistently showed higher levels of P-value inflation than burden tests. We determined that the differential stratification was caused by two specific patterns in the interpopulation distribution of rare variants, each correlating with inflation in one of the test classes. The pattern that inflates joint tests is more prevalent in real data, explaining the higher levels of inflation in these tests. Furthermore, we show that the different sources of inflation between tests lead to heterogeneous responses to genomic control correction and the number of variants analyzed. Our results indicate that care must be taken when interpreting joint and burden analyses of the same set of rare variants, in particular, to avoid mistaking inflated P-values in joint tests for stronger signals of true associations.


The sorption of influenza virus by chicken erythrocytes.
Data are presented which indicate that the forces of attraction between influenza virus and chicken erythrocytes are governed by an orderly mechanism which effects a proportional distribution of virus between erythrocytes and suspending fluid. For a unit mass of erythrocytes, the ratio of combined (sorbed) virus to free (unsorbed) virus is sufficiently constant over a wide range of virus concentrations to indicate compliance with the laws of mass action, or, perhaps, with the laws of simple adsorption; however, the ratio of combined (sorbed) virus to free (unsorbed) virus appears to be a parabolic function of the total quantity of virus.


Regulation of thrombin-induced stress fibre formation in Swiss 3T3 cells by the 70-kDa S6 kinase.
The signal transduction systems that mediate growth factor receptor-induced cellular shape change have not been fully elucidated, but are known to involve alterations in the state of actin filaments, termed stress fibres. It now appears from several studies that the GTP-binding protein, Rho, is involved. However, the mechanisms by which Rho is activated, and what effectors Rho in turn stimulates are largely matters of conjecture. The present work shows that thrombin is an effective stimulant of stress fibre formation in Swiss 3T3 cells. In addition, we show the 70 kDa form of S6 kinase (p70s6k) to colocalise with stress fibres in both unstimulated and thrombin-activated cells. Coincident with the thrombin-induced formation of stress fibres is the elevated association p70s6k with the fibres. Pretreatment of cells with rapamycin, to inhibit p70s6k activation, inhibits thrombin-induced stress fibre formation and the associated presence of p70s6k on the fibres, supporting a role for p70s6k in thrombin-stimulated stress fibre formation. Thrombin is also shown to stimulate p70s6k activity and that this is inhibited by rapamycin. Thus, the data presented show that thrombin activates stress fibre formation through stimulation of p70s6k via a non-Gi pathway.


Domestic violence against women: representations of health professionals.
to analyze the representations about domestic violence against women, among health professionals of Family Health Units. qualitative study based on the Theory of Social Representations. Data were collected by means of evocations and interviews, treating them in the Ensemble de Programmes Pemettant L'Analyse des Evocations software - EVOC and content analysis. nurses, physicians, nursing technicians and community health agents participated. The evocations were answered by 201 professionals and, of these, 64 were interviewed. The central core of this representation, comprised by the terms "aggression", "physical-aggression", "cowardice" and "lack of respect", which have negative connotations and were cited by interviewees. In the contrast zone, comprised by the terms "abuse", "abuse-power", "pain", "humiliation", "impunity", "suffering", "sadness" and "violence", two subgroups were identified. The first periphery contains the terms "fear", evoked most often, followed by "revolt", "low self-esteem" and "submission", and in the second periphery "acceptance" and "professional support". this is a structured representation since it contains conceptual, imagetic and attitudinal elements. The subgroups were comprised by professionals working in the rural area and by those who had completed their professional training course in or after 2004. These presented a representation of violence different from the representation of the general group, although all demonstrated a negative connotation of this phenomenon.


Experimental Methodology for Estimation of Local Heat Fluxes and Burning Rates in Steady Laminar Boundary Layer Diffusion Flames.
Modeling the realistic burning behavior of condensed-phase fuels has remained out of reach, in part because of an inability to resolve the complex interactions occurring at the interface between gas-phase flames and condensed-phase fuels. The current research provides a technique to explore the dynamic relationship between a combustible condensed fuel surface and gas-phase flames in laminar boundary layers. Experiments have previously been conducted in both forced and free convective environments over both solid and liquid fuels. A unique methodology, based on the Reynolds Analogy, was used to estimate local mass burning rates and flame heat fluxes for these laminar boundary layer diffusion flames utilizing local temperature gradients at the fuel surface. Local mass burning rates and convective and radiative heat feedback from the flames were measured in both the pyrolysis and plume regions by using temperature gradients mapped near the wall by a two-axis traverse system. These experiments are time-consuming and can be challenging to design as the condensed fuel surface burns steadily for only a limited period of time following ignition. The temperature profiles near the fuel surface need to be mapped during steady burning of a condensed fuel surface at a very high spatial resolution in order to capture reasonable estimates of local temperature gradients. Careful corrections for radiative heat losses from the thermocouples are also essential for accurate measurements. For these reasons, the whole experimental setup needs to be automated with a computer-controlled traverse mechanism, eliminating most errors due to positioning of a micro-thermocouple. An outline of steps to reproducibly capture near-wall temperature gradients and use them to assess local burning rates and heat fluxes is provided.


Ionic liquid-based approach to monodisperse luminescent LaF3:Ce,Tb nanodiskettes: synthesis, structural and photoluminescent properties.
In this paper, a novel and easy synthesis of highly luminescent, water-soluble LaF3:Ce,Tb nanodiskettes (25 nm in diameter) by ionic liquid-based hydrothermal process is reported. X-ray diffraction (XRD), transmission electron microscopy (TEM) and field emission scanning electron microscopy (FE-SEM) were used to characterize the structural properties of the LaF3:Ce,Tb nanodiskettes. Ionic liquid [bmim]BF4 acts as both a co-solvent and a reactant during the synthesis. The effects of EDTA-2Na on the formation of green LaF3:Ce,Tb nanodiskette phosphors were studied and discussed. The luminescent properties of LaF3:Ce,Tb nanodiskettes have been investigated under 254 nm excitation both in solid state and dispersed in solution. The quantum efficiency (in solid state) is about 20% compared with commercial green phosphor. The photoluminescent properties show that the as-prepared La0.8Ce0.15Tb0.05F3 nanodiskettes may be applied as phosphors for high-resolution displays and fluorescent labels for biomolecules.


Tris[diphenyl(2-pyridyl)phosphine]-micro(3)-iodo-tri-micro(3)-sulfido-sulfidotrisilvertungsten dichloromethane hemisolvate.
The title compound, [Ag(3)WIS(4)(C(17)H(14)NP)(3)].0.5CH(2)Cl(2), is a cubane-type heterometallic cluster containing diphenyl(2-pyridyl)phosphine (Ph(2)PPy). The pyridyl group of Ph(2)PPy remains uncoordinated, so the Ph(2)PPy ligand is monodentate and coordinates to one Ag atom. The W atom and three Ag atoms form a distorted tetrahedral geometry, capped by one I atom and three S atoms.


Delayed fluid resuscitation of head injury and uncontrolled hemorrhagic shock.
To evaluate the effects of delayed vs early fluid resuscitation on cerebral hemodynamics after severe head injury and uncontrolled hemorrhagic shock. Prospective, randomized, controlled experimental trial. Surgical research laboratory. Immature swine (N=16) weighing 40 to 50 kg. Twelve swine were subjected to cryogenic brain lesion and hemorrhage to maintain a mean arterial pressure (MAP) of 50 mm Hg. Animals were randomized to receive 1 L of Ringer lactate solution in 20 minutes, starting 20 minutes after injury and hemorrhage, followed by 1 L of Ringer lactate solution in 30 minutes (ER group) (n=6), or no fluid resuscitation (DR group) (n=6). The 4 control animals underwent instrumentation only. The study ended 70 minutes after head injury and hemorrhage. Measurements of MAP, bilateral regional cerebral blood flow, serum hemoglobin level, systemic and regional cerebral oxygen delivery, and intracranial pressure performed at baseline and 20 (phase 1), 50 (phase 2), and 70 minutes (phase 3) after head injury and hemorrhage. Lesion size (percentage of ipsilateral cortex) was measured post mortem. All animals survived the experimental period. Systemic cerebral oxygen delivery in the DR group was significantly lower at phase 3 compared with that of the ER group (31.5% vs 53.1% at baseline) (P=.03). However, bilateral regional cerebral oxygen delivery was significantly greater in the DR group at phase 3 compared with that of the ER group (71.5% vs 47.0% at baseline in the injured side; 72.9% vs 48.4% at baseline in the noninjured side) (P=.02). Bilateral cerebral blood flow was similar in all groups at all times. The ER group showed a trend toward a greater intracranial pressure elevation (6.8 vs -0.25) (P=.07) and lesion size (37.0% vs 28.6%) (P=.07). Hemoglobin level became significantly lower in the ER group at phase 2 (7.0 vs 10.7) (P=.03) and remained lower at phase 3 (6.9 vs 11.7) (P=.01). Early fluid resuscitation with Ringer lactate solution following head injury and uncontrolled hemorrhagic shock worsens cerebral hemodynamics. Cerebral pressure autoregulation is sufficiently intact following head injury to maintain regional cerebral oxygen delivery without asanguineous fluid resuscitation.


Touch Surgery: Analysis and Assessment of Validity of a Hand Surgery Simulation "App".
Surgical educators are increasingly exploring surgical simulation and other nonclinical teaching adjuncts in the education of trainees. The simulators range from purpose-built machines to inexpensive smartphone or tablet-based applications (apps). This study evaluates a free surgery module from one such app, Touch Surgery, in an effort to evaluate its validity and usefulness in training for hand surgery procedures across varied levels of surgical experience. Participants were divided into 3 cohorts: fellowship-trained hand surgeons, orthopedic surgery residents, and medical students. Participants were trained in the use of the Touch Surgery app. Each participant completed the Carpal Tunnel Release module 3 times, and participants' score was recorded for each trial. Participants also completed a customized Likert survey regarding their opinions on the usefulness and accuracy of the app. Statistical analysis using a 2-tailed t test and analysis of variance was performed to evaluate for performance within and between cohorts. All cohorts performed better on average with each subsequent simulation attempt. For all attempts, the experts outperformed the novice and intermediate participants, while the intermediate cohort outperformed the novice cohort. Novice users consistently gave the app better scores for usefulness as a training tool, and demonstrated more willingness to use the product. The study confirms app validity and usefulness by demonstrating that every cohort's simulator performance improved with consecutive use, and participants with higher levels of training performed better. Also, user confidence in this app's veracity and utility increased with lower levels of training experience.


Olfactoretinal centrifugal input modulates zebrafish retinal ganglion cell activity: a possible role for dopamine-mediated Ca2+ signalling pathways.
The vertebrate retina receives centrifugal input from the brain. In zebrafish, the major centrifugal input originates in the terminal nerve (TN). TN cell bodies are located in the olfactory bulb and ventral telencephalon. The TN projects axons to the retina where they branch in the inner plexiform layer (IPL) and synapse onto several inner retinal cell types, including dopaminergic interplexiform cells (DA-IPCs). This olfactoretinal centrifugal input plays a role in modulating retinal ganglion cell (RGC) activity, probably via dopamine-mediated Ca2+ signalling pathways. Normally, dopamine inhibits RGC firing by decreasing the inward Ca2+ current. Olfactory stimulation with amino acids decreases dopamine release in the retina, thereby reducing dopaminergic inhibition of RGCs. This model of olfacto-visual integration was directly tested by recording single-unit RGC activity in response to olfactory stimulation in the presence or absence of dopamine receptor blockers. Stimulation of the olfactory neurones increased RGC activity. However, this effect diminished when the dopamine D1 receptors were pharmacologically blocked. In isolated RGCs, the application of dopamine or a dopamine D1 receptor agonist decreased voltage-activated Ca2+ current and lowered Ca2+ influx. Together, the data suggest that olfactory input has a modulatory effect on RGC firing, and that this effect is mediated by dopamine D1 receptor-coupled Ca2+ signalling pathways.


Feasibility and safety of intrathecal treatment with nusinersen in adult patients with spinal muscular atrophy.
Nusinersen is an intrathecally administered antisense oligonucleotide (ASO) and the first approved drug for the treatment of spinal muscular atrophy (SMA). However, progressive neuromyopathic scoliosis and the presence of spondylodesis can impede lumbar punctures in SMA patients. Our aim was to assess the feasibility and safety of the treatment in adults with SMA. For the intrathecal administration of nusinersen, we performed conventional, fluoroscopy-assisted and computer tomography (CT)-guided lumbar punctures in adult patients with type 2 and type 3 SMA. We documented any reported adverse events and performed blood tests. We treated a total of 28 adult SMA patients (9 patients with SMA type 2 and 19 patients with SMA type 3) aged between 18-61 years with nusinersen. The mean Revised Upper Limb Module (RULM) score at baseline in SMA type 2 and SMA type 3 patients was 9.9 ± 4.6 and 29.5 ± 8.5, respectively. The mean Hammersmith Functional Motor Scale Expanded (HFMSE) score at baseline was 3.1 ± 2.5 and 31.2 ± 18.1, respectively. Half of the SMA type 3 patients were ambulatory at treatment onset. In total, we performed 122 lumbar punctures with 120 successful intrathecal administrations of nusinersen. Lumbar punctures were well tolerated, and no serious adverse events occurred. Our data demonstrate the feasibility and tolerability of intrathecal treatment with nusinersen in adults with SMA type 2 and type 3. However, treatment can be medically and logistically challenging, particularly in patients with SMA type 2 and in patients with spondylodesis.


Rabbit models of arthritis: immunolocalization of matrix metalloproteinases and tissue inhibitor of metalloproteinase in synovium and cartilage.
The distribution of the matrix metalloproteinases, collagenase, stromelysin, gelatinases A and B, and the tissue inhibitor of metalloproteinases in cartilage and synovium removed from rabbits up to 27 days after induction of two models of arthritis was investigated by immunolocalization. Following intra-articular injection of poly-D-lysine/hyaluronic acid coacervate, collagenase and stromelysin were found bound to cartilage matrix, but there was little increase in chondrocyte synthesis of these enzymes. The synovium underwent a complex wound healing response involving invagination and encapsulation of the coacervate and inflammatory cell debris, during which all four metalloproteinases and tissue inhibitor of metalloproteinase could be immunolocalized. The second model, intra-articular injection of ovalbumin into sensitized rabbits, caused considerable chondrocyte necrosis; collagenase was found bound to cartilage matrix on day 13, although again there was little evidence of synthesis by chondrocytes. Inflammatory cell infiltration of meniscoid synovia took place initially, followed by fibrosis involving macrophagelike cells secreting gelatinase A. In both models there was rapid loss of glycosaminoglycan metachromasia from the cartilage matrix. These results are discussed in relation to current knowledge of metalloproteinase involvement in the chronic rheumatoid synovial pannus erosion of cartilage in humans. The data suggest that there are considerable differences between rheumatoid arthritis and these models, and their use must therefore be carefully defined.


Persistence and outcomes of non-vitamin K antagonist oral anticoagulants versus warfarin in patients with non-valvular atrial fibrillation.
To compare persistence and outcomes of non-vitamin K antagonist oral anticoagulants (NOACs) versus warfarin in Chinese patients with non-valvular atrial fibrillation (AF). Given the unpredictable warfarin response and the costliness of NOACs, more research is needed to clarify which drug enjoys better persistence and outcomes, helping to provide personalised care for patients. A prospective cohort study. Chinese patients taking NOACs or warfarin from March 2016-April 2018 were followed up by telephone or outpatient visit at 3, 6 months and half a year thereafter. Anticoagulant persistence and outcomes including stroke and bleeding were collected. We used Cox regression to analyse data. This study was reported according to the STROBE guideline. A total of 344 patients were enrolled; 146 patients received NOACs including dabigatran and rivaroxaban, and 198 patients received warfarin. Persistence with anticoagulants was low and dropped sharply at the third month. Patients on NOACs had worse persistence at 3, 6 and 12 months than those on warfarin. There was no difference in the incidence of ischaemic stroke and bleeding between groups, although ischaemic stroke and major bleeding occurred less frequently in the NOACs group. Paroxysmal AF, no heart failure and no stroke were predictors of NOACs non-persistence. Prior catheter ablation and no diabetes were associated with poor persistence of warfarin. The main reason for anticoagulant cessation was patient preference. Chinese patients taking NOACs had lower persistence, similar rate of ischaemic stroke and bleeding compared with those on warfarin. Further inventions are needed to improve persistence in Chinese patients on NOACs. Anticoagulation should highlight both persistence and outcomes emphasising personalised care of different drugs. Further interventions to improve persistence should be developed based on causes and risk factors and carried out in the third month of therapy.


Anal Melanoma in an Elderly Woman Masquerading as Hemorrhoid.
Anal melanoma is a rare and aggressive neoplasm of the anal canal seen in the elderly population in the six or seventh decade of their lives. Presentation is usually nonspecific and diagnosis is often delayed or missed initially. The management is surgical and prognosis is poor. Here we present a case of anal melanoma in an elderly patient masquerading as hemorrhoid.


Vanadium Chloroperoxidases: The Missing Link in the Formation of Chlorinated Compounds and Chloroform in the Terrestrial Environment?
It is well established that the majority of chlorinated organic substances found in the terrestrial environment are produced naturally. The presence of these compounds in soils is not limited to a single ecosystem. Natural chlorination is also a widespread phenomenon in grasslands and agricultural soils typical for unforested areas. These chlorinated compounds are formed from chlorination of natural organic matter consisting of very complex chemical structures, such as lignin. Chlorination of several lignin model compounds results in the intermediate formation of trichloroacetyl-containing compounds, which are also found in soils. These decay, in general, through a haloform-type reaction mechanism to CHCl3 . Upon release into the atmosphere, CHCl3 will produce chlorine radicals through photolysis, which will, in turn, lead to natural depletion of ozone. There is evidence that fungal chloroperoxidases able to produce HOCl are involved in the chlorination of natural organic matter. The objective of this review is to clarify the role and source of the various chloroperoxidases involved in the natural formation of CHCl3 .


[Limited access to the international medical literature in Russia].
Limited access to foreign professional literature in the former Soviet Union had consequences for public health: persistence of some outdated methods and approaches. Several examples are discussed in this letter. The shortage of foreign literature has been partly compensated by domestic editions, sometimes containing compilations from foreign sources, borrowings without references, and mistranslations. International literature is on average scarcely quoted in Russian language scientific publications. Today, however, there are grounds for optimism: the economic upturn must bring improvements.


Histological classification of canine ovarian cyst types with reference to medical history.
Ovaries of 21 bitches presented with gynecopathies were surgically removed and histologically examined. Standard histological, as well as immunohistochemical, classification of 193 cystic structures resulted in the classification of 72 cysts of subsurface epithelial structures (SES), 61 follicular cysts (FCs), 38 cystic rete ovarii (CRO), 13 lutein cysts (LCs), and 9 non-classifiable cysts (NCCs). In addition to the histological classification, results were interpreted according to subject medical history, clinical examination outcome, and macroscopic observations during ovariohysterectomy. Dogs with ovarian cysts (OCs) and associated reproductive perturbations were mostly nulliparous, of large breed, and had an average of 9.5 ± 3 years. Prolonged or shortened inter-estrus intervals of past heats, however, seemed to be relatively low-risk factors for the development of OCs in dogs. Furthermore, we provide histological observations of a rarely seen canine LC including a degenerated oocyte in the central cavity.


Forkhead Protein FoxO1 Acts as a Repressor to Inhibit Cell Differentiation in Human Fetal Pancreatic Progenitor Cells.
Our colleagues have reported previously that human pancreatic progenitor cells can readily differentiate into insulin-containing cells. Particularly, transplantation of these cell clusters upon in vitro induction for 3-4 w partially restores hyperglycemia in diabetic nude mice. In this study, we used human fetal pancreatic progenitor cells to identify the forkhead protein FoxO1 as the key regulator for cell differentiation. Thus, induction of human fetal pancreatic progenitor cells for 1 week led to increase of the pancreatic β cell markers such as Ngn3, but decrease of stem cell markers including Oct4, Nanog, and CK19. Of note, FoxO1 knockdown or FoxO1 inhibitor significantly upregulated Ngn3 and insulin as well as the markers such as Glut2, Kir6.2, SUR1, and VDCC, which are designated for mature β cells. On the contrary, overexpression of FoxO1 suppressed the induction and reduced expression of these β cell markers. Taken together, these results suggest that FoxO1 may act as a repressor to inhibit cell differentiation in human fetal pancreatic progenitor cells.


Proteomic Signatures of Microbial Adaptation to the Highest Ultraviolet-Irradiation on Earth: Lessons From a Soil Actinobacterium.
In the Central Andean region in South America, high-altitude ecosystems (3500-6000 masl) are distributed across Argentina, Chile, Bolivia, and Peru, in which poly-extremophilic microbes thrive under extreme environmental conditions. In particular, in the Puna region, total solar irradiation and UV incidence are the highest on Earth, thus, restraining the physiology of individual microorganisms and the composition of microbial communities. UV-resistance of microbial strains thriving in High-Altitude Andean Lakes was demonstrated and their mechanisms were partially characterized by genomic analysis, biochemical and physiological assays. Then, the existence of a network of physiological and molecular mechanisms triggered by ultraviolet light exposure was hypothesized and called "UV-resistome". It includes some or all of the following subsystems: (i) UV sensing and effective response regulators, (ii) UV-avoidance and shielding strategies, (iii) damage tolerance and oxidative stress response, (iv) energy management and metabolic resetting, and (v) DNA damage repair. Genes involved in the described UV-resistome were recently described in the genome of Nesterenkonia sp. Act20, an actinobacterium which showed survival to high UV-B doses as well as efficient photorepairing capability. The aim of this work was to use a proteomic approach together with photoproduct measurements to help dissecting the molecular events involved in the adaptive response of a model High-Altitude Andean Lakes (HAAL) extremophilic actinobacterium, Nesterenkonia sp. Act20, under artificial UV-B radiation. Our results demonstrate that UV-B exposure induced over-abundance of a well-defined set of proteins while recovery treatments restored the proteomic profiles present before the UV-challenge. The proteins involved in this complex molecular network were categorized within the UV-resistome subsystems: damage tolerance and oxidative stress response, energy management and metabolic resetting, and DNA damage repair.


A novel mutation in the RSPO4 gene in a patient with autosomal recessive anonychia.
The Wnt signalling pathway is a major pathway involved in the embryogenic development of the various organs of the body. Appropriate signalling in this pathway relies on the proper functioning of several proteins including the R-spondin family of proteins. Deactivating mutations in R-spondin 4 are associated with anonychia. We present the case of a 26-year-old man presenting with anonychia of the 20 nails, which had been present since birth. Using genetic studies, we identified a novel nonsense mutation, c.164-165TC>AA, characterized by two consecutive mismatch bases. To our knowledge, this mutation is the first to be reported in R-spondin 4 in a Lebanese population. Evaluating new patients with anonychia provides fruitful clinical and molecular findings.


Stereotactic large-gauge core biopsy: its role in the diagnosis of non-palpable mammographic abnormalities presenting to a screening service.
A retrospective study of large-gauge core biopsy (LGCB) of 206 non-palpable mammographic abnormalities detected at routine screening in a community-based clinic accredited with the Australian National Program for the Early Detection of Breast cancer, was performed to ascertain the role of LGCB as an alternative to diagnostic surgical biopsy within such a programme. Of the 51 malignancies diagnosed by LGCB and proceeding to treatment, no false-positives were found and agreement as to the presence, or otherwise, of invasion between core and open biopsy was 95%. Once malignancy was established by LGCB, one-stage treatment was possible in 74.5% of cases. A total of 114 lesions were considered to be benign and returned to routine screening. One interval cancer subsequently developed in this group. Incongruity between the pathological diagnosis from core biopsy and the mammographic image occurred in 29 cases, representing potential mistargeting in 14.1% of cases. Four cancers were discovered in this group when the patients subsequently proceeded to surgical biopsy, making an overall false-negative rate of 2.4%. No core biopsy sample was considered inadequate for pathological assessment and no lesion proved inaccessible to targeting. There was no long-term morbidity. We believe that LGCB is a safe, reliable and cheaper alternative to diagnostic surgical biopsy, but a close correlation of pathology and mammography is required to avoid the erroneous return of women to routine recall.


Excessive Surgical Adhesive Mimicking Aortic Root Abscess: A Case Report.
Aortic root abscess is a complication of aortic valve endocarditis that is associated with a high morbidity and mortality. The diagnosis usually is made with transesophageal echocardiography, which is highly sensitive and specific for the disease. We present a case of suspected aortic root abscess 1 week after mechanical aortic valve replacement for native valve endocarditis. The diagnosis was made by the use of transesophageal echocardiography but surgical inspection revealed that the paravalvular fluid collection was excessive surgical adhesive. We discuss the clinical significance and differential diagnosis of aortic root abscess in the setting of infective endocarditis.


Discussion of the sensory innervation of the brain.
In response to a commentary from another author, the present investigator provides new data obtained in his laboratory on the similarity of a number of brain neurons bearing asynaptic dendrites and innervating the perivascular space, the pia mater (from within), and making direct contact with the cerebrospinal fluid in the ventricles on the one hand with local intramural autonomic Dogiel type II neurons in the internal organs on the other. Our own electron microscopy data on the terminals of asynaptic dendrites in the brain are presented, these showing no difference in terms of ultrastructure from the terminals of receptors in the internal organs. Factual data are presented which, in the author's opinion, refute the other author's views that cilia in general and ciliated neurons in particular cannot have a variety of receptor functions, that sensory neurons arise exclusively from the neural crest, and that it is fundamentally impossible for brain tissues to have sensory innervation.


Evaluation of the Mini-ID Enterobacteriaceae screen system.
A total of 932 typical and atypical enteric fermenters were used to evaluate the Mini-ID Enterobacteriaceae Screen/Identification System. At 4 h, final identifications were available for only 13.3%, but an additional 71% were screened into the correct group, according to the product's database. At 24 h, 58.8% were correctly identified to the species level, often with the use of an additional tube. When 118 of the cultures were arranged into a weighted assortment, as might be found in a clinical laboratory, 35 were definitively identified at 4 h, and another 72 were screened into the correct group. Of these 72, 31 were correctly identified to the species level at 24 h, for a total of 56.0%. False-negative ornithine and incorrect L-pyrrolidonyl-beta-naphthylamide and glucuronide xylopyranoside reactions accounted for 54% of the identification errors, while database problems accounted for 10.2% of the errors. Of the eight Salmonella paratyphi A cultures, seven were missed because of a false-positive lysine reaction. At best, the system serves only as a rough screen.


Genetic screening for cystic fibrosis.
More than 98% of mutations causing cystic fibrosis can be detected in a Celtic population in Brittany, France. What, though, are the prospects for screening of entire populations for carriers?... Even if technology improves to the point where 95% of CF mutations can be detected routinely, and population screening becomes feasible, a more contentious matter is that of counselling and education, especially as there is every reason to believe that the lifespan of CF patients will continue to grow as therapies improve....


3H-1,2-benzoxathiepine 2,2-dioxides: a new class of isoform-selective carbonic anhydrase inhibitors.
A new chemotype with carbonic anhydrase (CA, EC 126.96.36.199) inhibitory action has been discovered, the homo-sulfocoumarins (3H-1,2-benzoxathiepine 2,2-dioxides) which have been designed considering the (sulfo)coumarins as lead molecules. An original synthetic strategy of a panel of such derivatives led to compounds with a unique inhibitory profile and very high selectivity for the inhibition of the tumour associated (CA IX/XII) over the cytosolic (CA I/II) isoforms. Although the CA inhibition mechanism with these new compounds is unknown for the moment, we hypothesize that it may be similar to that of the sulfocoumarins, i.e. hydrolysis to the corresponding sulfonic acids which thereafter anchor to the zinc-coordinated water molecule within the enzyme active site.


Pharmacokinetics and Disposition of Contezolid in Humans: Resolution of a Disproportionate Human Metabolite for Clinical Development.
Contezolid (MRX-I), a novel oxazolidinone antibiotic, was recently approved for the treatment of serious Gram-positive infections. The pharmacokinetics and disposition of [14C]contezolid were investigated in a single-dose human mass balance study. Cross-species comparison of plasma exposure for contezolid and metabolites was performed, and the safety of the disproportionate metabolite in human was evaluated with additional nonclinical studies. After an oral administration of 99.1 μCi/602-mg dose of [14C]contezolid, approximately 91.5% of the radioactivity was recovered in 0 to 168 h postdose, mainly in urine followed by that in feces. The principal metabolic pathway of contezolid in human comprised an oxidative ring opening of the 2,3-dihydropyridin-4-one fragment into polar metabolites MRX445-1 and MRX459, with recovery of approximately 48% and 15% of the dose, respectively, in urine and feces. Contezolid, MRX445-1, and MRX459 accounted for 68.0%, 19.5%, and 4.84% of the plasma exposure of the total radioactivity, respectively. Metabolites MRX445-1 and MRX459 were observed in disproportionately larger amounts in human plasma than in samples from rat or dog, the rodent and nonrodent species, respectively, used for the general nonclinical safety assessment of this molecule. This discrepancy was resolved with additional nonclinical studies, wherein the primary metabolite, MRX445-1, was further characterized. The no-observed-adverse-effect level (NOAEL) of MRX445-1 was determined as 360 mg/kg body weight/day in a 14-day repeat-dose test in pregnant and nonpregnant Sprague Dawley rats. Furthermore, MRX445-1 exhibited no antibacterial activity in vitro. Thus, MRX445-1 is not expected to exert clinically relevant pharmacology and toxicity.


Chromophore formation in resorcinarene solutions and the visual detection of mono- and oligosaccharides.
The colorimetric properties of resorcinarene solutions had not been investigated since Baeyer's initial synthesis. We recently reported that solutions containing resorcinarene macrocycles develop color upon heating or standing. In the presence of saccharides, these solutions exhibit significant color changes which are easily seen. We herein present strong evidence that the solution color is due to macrocycle ring opening and oxidation. The optical responses to saccharides are due to complexation of the sugar with the acyclic chromophores. We apply these mechanistic insights toward the challenging problem of the visual detection of neutral oligosaccharides by simple chromogens. In addition, we also report the first single-crystal X-ray crystal structure determination of a rarely observed "diamond" resorcinarene stereoisomer.


Thoracic organ transplantation in children. The state of heart, heart-lung, and lung transplantation.
Heart, heart-lung, and lung transplantation have become accepted modalities for treatment in children with serious cardiopulmonary disease. Although early deaths secondary to infection and/or acute rejection have been reduced dramatically, there is still an early mortality related to cardiac complications and a late mortality related to rejection and infection. The management of hypoplastic left heart syndrome remains a special problem and is outlined in the review. There is much less experience in the newer modalities of heart-lung and lung transplantation in children and the results of these operative procedures are similar to what was seen in heart transplantation some 10 years ago. Ongoing and significant improvement continues, however, in all forms of transplantation dealing with the lung.


Transactivation of methylated HIV-LTR by a frog virus 3 protein.
DNA methylation has been implicated in the suppression of transcription of a large spectrum of eukaryotic genes. Frog virus 3 (FV3) contains genomic DNA that is the most extensively methylated of all known animal viruses. However, FV3 gene expression is tightly regulated in a sequential fashion in infected cells. Therefore, FV3 must have evolved a mechanism(s) to overcome the inhibitory effects of DNA methylation. FV3 has been shown to induce expression of methylated foreign genes in transient transfections. This study was designed to establish if this FV3-induced expression of methylated genes could be demonstrated in stable cell lines which contain integrated foreign genes that are silenced by DNA methylation. Stably transfected simian Vero and human T-cells containing a single copy of the methylated and transcriptionally suppressed HIV-LTR CAT construct were either infected with FV3 or fused with FV3-infected fat head minnow cells. The results from these experiments lead us to conclude that FV3 infection does promote expression of a foreign, stably integrated gene (HIV-LTR), which was previously silenced by DNA methylation. We also observed that stably transformed human T-cells incubated at 30 degrees, unlike at 37 degrees, expressed minute but detectable HIV-LTR-directed CAT activity. Significance of this finding in HIV pathogenesis remains elusive.


The effect of the disaster caused by the great Hanshin earthquake on people with intellectual disability.
The vulnerability of people with intellectual disability when services are disrupted by a major earthquake is described. The value of an accessible register and multiple networks of support at the time of a major disaster is emphasized.


Utilizing a cranial window to visualize the middle cerebral artery during endothelin-1 induced middle cerebral artery occlusion.
Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain(1-3). This technique can be performed in many locations overlying the rat cerebrum, but is most easily carried out by creating a craniectomy over the readily accessible frontal or parietal bones. Most frequently, we have used this technique in combination with the endothelin-1 middle cerebral artery occlusion model of ischemic stroke to quantify the changes in middle cerebral artery vessel diameter that occur with injection of endothelin-1 into the brain parenchyma adjacent to the proximal MCA(4, 5). In order to visualize the proximal portion of the MCA during endothelin -1 induced MCAO, we use a technique to create a cranial window through the temporal bone on the lateral aspect of the rat skull (Figure 1). Cerebral arteries can be visualized either with the dura intact or with the dura incised and retracted. Most commonly, we leave the dura intact during visualization since endothelin-1 induced MCAO involves delivery of the vasoconstricting peptide into the brain parenchyma. This bypasses the need to incise the dura directly over the visualized vessels for drug delivery. This protocol will describe how to create a cranial window to visualize cerebral arteries in a step-wise fashion, as well as how to avoid many of the potential pitfalls pertaining to this method.


Experience Using Ustekinumab in Pediatric Patients With Medically Refractory Crohn Disease.
Ustekinumab (UST), a human monoclonal antibody against interleukin-12 and 23, is approved to treat adult patients with psoriasis or Crohn disease (CD). Outcomes data for off-label use in pediatric patients with CD are limited. We conducted a retrospective cohort study to analyze the long-term efficacy of UST, including dose adjustments, in the treatment of pediatric patients with medically refractory CD. Adverse events were documented. We identified 40 pediatric patients with CD treated with UST between January 1, 2016 and December 31, 2019. Electronic medical records were reviewed for demographics, Paris Classification, significant comorbidities, previous CD therapy, adverse events after initiation, and surveillance markers at the time of their first dose and most recent clinic visit. A validated abbreviated pediatric CD activity index (aPCDAI) was used to assess response to therapy. Thirty-eight pediatric patients with CD, including 34.2% with stricturing or penetrating disease, were analyzed after initiation of treatment with UST. Median age at diagnosis of CD was 12.5 years, and median age at UST induction was 17.2 years. No patients were anti-TNF-naive, and 34.2% were previously exposed to 2 or more anti-TNF agents. At time of last follow-up, 84.2% of patients remained on UST for a median duration on UST of 62.1 weeks, and 60.5% achieved clinical remission. Patients had significant improvement in aPCDAI scores, clinical remission rates, albumin, and hematocrit, and 89.5% of patients had no significant adverse events. Similar results were observed among those who required dose adjustment, including 61.1% achieving clinical remission, and among those with perianal disease, including 38.5% achieving clinical remission. Our data suggest that, within our cohort of pediatric patients with CD, UST has long-term efficacy with no observed safety concerns. Dose adjustment may be helpful in achieving clinical remission.


Evaluation of implementation status of national policy on Indian systems of medicine and homeopathy 2002: Stakeholders' perspective.
National Policy on Indian systems of medicine and homoeopathy (ISM and H policy) was formulated in 2002 to encourage the development of Ayurveda, Sidhha, Unani, Yoga, Naturopathy and Homoeopathy in India. This study proposes to assess the views of public health experts on current implementation of ISM and H Policy. An online questionnaire was designed to ascertain the views of public health experts on ISM and H Policy and mailed to 100 public health experts. The tool was tested for content validity and a pilot study was done. Results were analyzed with the help of SPSS version 16. Response rate was 61%. Majority of experts considered implementation status of ISM and H Policy as poor. Lack of quality education was mentioned as major factor responsible for current scenario of ISM and H Policy by most of experts. Lack of funds and government support were emerged as major bottlenecks in implementation of ISM and H Policy.


A pilot study on the changes of event-related potentials in school-aged children with iron deficiency anaemia.
Event-related potentials (ERP) were assessed in 70 school-aged children with a diagnosis of asymptomatic iron deficiency anaemia (IDA), based on low haemoglobin and either low serum ferritin or high free erythrocyte protoporphyrin levels. The IDA subjects were randomized into treatment and placebo groups of 35 cases each, and compared with a normal control group of 30 age- and gender-matched healthy subjects without iron deficiency. Further haematological and ERP assessment was carried out after 3 months, during which time the active group received iron supplementation with 10 mg ferrous sulphate, together with vitamin C, malic acid and folic acid. Pre-treatment, both IDA groups had prolonged P300 latencies in comparison with the non-IDA controls (p<0.01). The proportion of cases with distorted wave appearance was more than twice as high in the IDA groups as in the non-IDA controls, although intergroup differences did not reach statistical significance. After treatment, the active treatment IDA group showed a significant increase in haemoglobin levels and shortening in P300 latencies. After treatment, neither value was statistically different from non-IDA controls. There was a decrease in the number of cases with abnormal waveforms in the active treatment group, compared with an increase in the number within the placebo group (P = 0.002). Testing of ERP shows promise as a non-invasive, sensitive and objective marker for assessing cognitive impairment in children with IDA.


Haptic Feedback in Needle Insertion Modeling and Simulation.
Needle insertion is the most basic skill in medical care, and training has to be imparted not only for physicians but also for nurses and paramedics. In most needle insertion procedures, haptic feedback from the needle is the main stimulus in which novices need training. For better patient safety, the classical methods of training the haptic skills have to be replaced with simulators based on new robotic and graphics technologies. This paper reviews the current advances in needle insertion modeling, classified into three sections: needle insertion models, tissue deformation models, and needle-tissue interaction models. Although understated in the literature, the classical and dynamic friction models, which are critical for needle insertion modeling, are also discussed. The experimental setup or the needle simulators that have been developed to validate the models are described. The need of psychophysics for needle simulators and psychophysical parameter analysis of human perception in needle insertion are discussed, which are completely ignored in the literature.


Fibroblast activating protein-α expression in squamous cell carcinoma of the esophagus in primary and irradiated tumors: the use of archival FFPE material for molecular techniques.
There are numerous reports suggesting that fibroblast activating protein-α (FAP-α) plays an important role in invasion of various tumor types. We studied the expression pattern of FAP-α in esophageal squamous cell carcinoma (ESCC) patients who had not been treated primarily and those who had received neoadjuvant radiochemotherapy. Our goal was to establish whether readily available tissue specimens fixed in formalin and stored in paraffin blocks for years might still be a source of FAP-α RNA for PCR analysis. The study included 20 patients divided into two groups, 10 patients in each group. We evaluated the expression of FAP-α by PCR techniques in fresh frozen and in paraffin-embedded tissues, and compared it to the expression in non-cancer tissues. To detect the protein expression level of FAP-α in paraffin-embedded tissues we used chromogenic immunohistochemical (IHC) staining. Data were analyzed by t-test or the nonparametric Wilcoxon matched pair test using Statistica 12.5 software. We observed an increased level of the FAP-alpha gene and protein expression in cancer tissues when compared with their corresponding normal tissues. However, statistically significant differences were found only in the group of patients untreated before surgery. RNA extracted from paraffin-embedded tissue sections had very low quality, especially in the context of degradation. FAP-α remains a highly altered participant of a complex microenvironment in esophageal squamous cell carcinoma, and its role in cell signaling requires further study. In this paper, we conclude that the use of a regular RT-PCR method for diagnostic purposes, which we have presented in an earlier paper, can be as good as qRT-PCR. Also, immunohistochemistry proved to be very useful and the only reliable method that can be used on formalin-fixed, paraffin-embedded tissues stored long term.


Invasive Ocular Surface Squamous Neoplasia Masquerading as Nodular Scleritis.
The authors report a rare case of ocular surface squamous neoplasia with intraocular involvement that had an initial masquerade presentation of recurrent anterior nodular scleritis. A 35-year-old male patient presented with right eye recurrent anterior nodular scleritis for which a lamellar patch graft was done. Two months later, the patient presented with recurrence of symptoms. Histopathology review revealed the presence of well-differentiated squamous cell malignancy. A high index of suspicion for malignancy is required in such cases when they do not respond to conventional therapy.


Enantioseparation of cetirizine by chromatographic methods and discrimination by 1H-NMR.
Cetirizine is an antihistaminic drug used to prevent and treat allergic conditions. It is currently marketed as a racemate. The H1-antagonist activity of cetirizine is primarily due to (R)-levocetirizine. This has led to the introduction of (R)-levocetirizine into clinical practice, and the chiral switching is expected to be more selective and safer. The present work represents three methods for the analysis and chiral discrimination of cetirizine. The first method was based on the enantioseparation of cetirizine on silica gel TLC plates using different chiral selectors as mobile phase additives. The mobile phase enabling successful resolution was acetonitrile-water 17: 3, (v/v) containing 1 mM of chiral selector, namely hydroxypropyl-beta-cyclodextrin, chondroitin sulphate or vancomycin hydrochloride. The second method was a validated high performance liquid chromatography (HPLC), based on stereoselective separation of cetirizine and quantitative determination of its eutomer (R)-levocetirizine on a monolithic C18 column using hydroxypropyl-beta-cyclodextrin as a chiral mobile phase additive. The resolved peaks of (R)-levocetirizine and (S)-dextrocetirizine were confirmed by further mass spectrometry. The third method used a (1)H-NMR technique to characterize cetirizine and (R)-levocetirizine. These methods are selective and accurate, and can be easily applied for chiral discrimination and determination of cetirizine in drug substance and drug product in quality control laboratory. Moreover, chiral purity testing of (R)-levocetirizine can also be monitored by the chromatographic methods.


Comparing the 810nm diode laser with conventional surgery in orthodontic soft tissue procedures.
To compare the use of the 810nm diode laser with conventional surgery in the management of soft tissue mucogingival problems associated with orthodontic treatment. Orthodontic patients requiring different soft tissue surgical procedures were randomly assigned to receive conventional surgery or soft tissue diode laser, (wavelength 810 nm). Parameters documented include the type of anaesthesia used, intra and post operative pain, bleeding, the use of scalpel and sutures. The chi-squared test was used to test for significance at 95% confidence level. Probability values (p-values) less than 0.05 were regarded as significant. Only 2(16.7%) of the procedures carried out with the soft tissue laser required infiltration anaesthesia compared to 10 (90.9%) with conventional surgery and this was significant (P<0.001). Post operative pain was significantly reduced in all cases treated with the diode laser (P<0.001). There was also a significant difference (P<0.05) in post operative bleeding in all cases treated with the diode laser. No sutures were used in all soft tissue cases managed with the diode laser and this was significant (P<0.001). There was no statistically significant difference in treatment time in the use of the laser compared with conventional surgery. Orthodontic patients treated with the diode laser required less infiltration anaesthesia, had reduced bleeding during and after surgery, rapid postoperative haemostasis, elimination of the need for sutures and an improved postoperative comfort and healing.


Model-based analysis on systemic availability of co-administered cannabinoids after controlled vaporised administration.
The most important two medicinal cannabinoids are Δ9 -tetrahydrocannabinol (THC) and cannabidiol (CBD). Vaporised administration is superior due to its higher systemic availability, lower individual variability and faster drug delivery. Although it is common THC is co-administered with CBD, the influence of CBD on the pharmacokinetics, especially the systemic availability of THC after vaporised administration, is unknown. To investigate the influence of different doses of co-administered CBD on the systemic availability of THC, and to compare the availability of THC and CBD in a sample of frequent and infrequent cannabis users. The study used a randomised, double-blind, crossover placebo-controlled design. THC and/or CBD in ethanol was vaporised and inhaled. Plasma concentrations of THC and CBD were analysed. The THC data created in this study were pooled together with published THC pharmacokinetic data in order to cover all the phases of THC disposition. Population pharmacokinetic model of THC was developed based on the pooled data. The model of CBD was developed based on the data created in this study. Population pharmacokinetic models of THC and CBD were developed. With concomitant inhalation of high-dose CBD, the systemic availability of THC decreased significantly. Frequent cannabis users appeared to have higher systemic availability of THC and CBD when high-dose CBD was administered. The results observed in this study are useful for guiding future pharmacokinetic studies of medicinal cannabinoids, and for development of dosing guidelines for medical use of cannabis in the 'real-world' setting.


Multiple sigma subunits and the partitioning of bacterial transcription space.
Promoter recognition in eubacteria is carried out by the initiation factor sigma, which binds RNA polymerase and initiates transcription. Cells have one housekeeping factor and a variable number of alternative sigma factors that possess different promoter-recognition properties. The cell can choose from its repertoire of sigmas to alter its transcriptional program in response to stress. Recent structural information illuminates the process of initiation and also shows that the two key sigma domains are structurally conserved, even among diverse family members. We use the sigma repertoire of Escherichia coli, Bacillus subtilis, Streptomyces coelicolor, and cyanobacteria to illustrate the different strategies utilized to organize transcriptional space using multiple sigma factors.


Full thickness skin graft versus split thickness skin graft in paediatric patients with hand burns: Systematic review and meta-analysis.
Our objective was to compare the outcomes of full thickness skin grafts versus split thickness skin grafts in paediatric hand burn patients. A systematic review and meta-analysis were carried out in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) Guidelines, and an electronic search was conducted to identify all Randomised Controlled Trials and non-randomised studies comparing the outcomes of full thickness skin grafts versus split thickness skin grafts in paediatric hand burn patients. Primary outcomes included development of post-graft contracture and the necessity for surgical release. Secondary outcomes consisted of evaluation of function, cosmesis and colour, scar and feeling, hair growth, and other complaints. For the analysis, fixed effects modelling was applied. Results: ten non-randomised trials with a total of 532 grafts were found. Full thickness skin grafts exhibited a statistically significant decrease in the development of post-graft contracture (Odds Ratio [OR] = 0.35, P = 0.0001) and later surgical releases (OR = 0.06, P = 0.00001). For secondary outcomes, full thickness skin grafts outperformed split thickness skin grafts in post-operative functional ability. However, split thickness skin grafts, showed to be superior in scar, aesthetic, and colour assessments, and less hair growth was observed for split thickness skin grafts. No significant difference was seen in sensation and donor or recipient site complaints. Overall, full thickness skin grafts are a better alternative for paediatric hand burns than split thickness skin transplants because they are linked with reduced post-graft contracture and the requirement for surgical release.


Involvement of PLA2, COX and LOX in Rhinella arenarum oocyte maturation.
In Rhinella arenarum, progesterone is the physiological nuclear maturation inducer that interacts with the oocyte surface and starts a cascade of events that leads to germinal vesicle breakdown (GVBD). Polyunsaturated fatty acids and their metabolites produced through cyclooxygenase (COX) and lipoxygenase (LOX) pathways play an important role in reproductive processes. In amphibians, to date, the role of arachidonic acid (AA) metabolites in progesterone (P4)-induced oocyte maturation has not been clarified. In this work we studied the participation of three enzymes involved in AA metabolism - phospholipase A2 (PLA2), COX and LOX in Rhinella arenarum oocyte maturation. PLA2 activation induced maturation in Rhinella arenarum oocytes in a dose-dependent manner. Oocytes when treated with 0.08 μM melittin showed the highest response (78 ± 6% GVBD). In follicles, PLA2 activation did not significantly induce maturation at the assayed doses (12 ± 3% GVBD). PLA2 inhibition with quinacrine prevented melittin-induced GVBD in a dose-dependent manner, however PLA2 inactivation did not affect P4-induced maturation. This finding suggests that PLA2 is not the only phospholipase involved in P4-induced maturation in this species. P4-induced oocyte maturation was inhibited by the COX inhibitors indomethacin and rofecoxib (65 ± 3% and 63 ± 3% GVBD, respectively), although COX activity was never blocked by their addition. Follicles showed a similar response following the addition of these inhibitors. Participation of LOX metabolites in maturation seems to be correlated with seasonal variation in ovarian response to P4. During the February to June period (low P4 response), LOX inhibition by nordihydroguaiaretic acid or lysine clonixinate increased maturation by up to 70%. In contrast, during the July to January period (high P4 response), LOX inhibition had no effect on hormone-induced maturation.


Matrixlysis, an improved sample preparation method for recovery of Mycobacteria from animal tissue material.
Mycobacterium caprae, a member of the Mycobacterium tuberculosis complex, is the main causative agent of bovine tuberculosis in alpine regions. Bacterial culture is the gold standard in bovine tuberculosis diagnostic but takes up to twelve weeks. This increases the time and costs for stocks affected with bovine tuberculosis. Hence this study focused on the implementation of a fast and precise mycobacterial detection method and compared it with currently used methods. Matrix lysis is a chemical lysis using high concentrations of urea to solubilize bovine and red deer tissue and was used to detect even smallest amounts or non-visible lesions of mycobacteria. A total of 64 samples collected from 44 animals (37 red deer and 7 cattle) were tested by Matrix lysis. Forty-three of these samples were used for Mycobacterium tuberculosis complex detection by quantitative PCR and other 21 for subtyping the genetically different variants of M. caprae. Furthermore, three Matrix lysis samples were used for Next Generation Sequencing. Our results confirm that Matrix lysis is a fast and precise method for detecting Mycobacterium tuberculosis complex in native tissue samples. However, at the moment it reaches its limits when the samples were analyzed by Next Generation Sequencing and RD4 subtyping.


Staphylococcus aureus enterotoxin mediated specific non-responsiveness of human T cells.
Exotoxins produced by certain strains of Staphylococcus aureus are able to both stimulate and induce non-responsiveness in T cells expressing specific T cell antigen receptor V beta gene elements. The exposure of human CD4+ T cells to the appropriate enterotoxin rendered them anergic to restimulation with their natural ligand, although responsiveness to exogenous IL-2 remained intact. The loss of antigen-dependent proliferation was associated with the down-regulation of the TCR complex that was paralleled by enhanced cell surface CD2 and CD25. Further analysis of the phenotypic changes revealed that membrane levels of CD28 were increased only on activation, suggesting a differential expression of this protein on activated and anergic T cells. During the induction of anergy it was observed that the synthesis of the lymphokines IL-2, IL-4 and IFN-gamma was differentially regulated. IL-4 and IFN-gamma, but not IL-2 were detected in the supernatants of overnight cultures of T cells exposed to tolerising concentrations of toxin. Transcription of IL-4, as determined by polymerase chain reaction at selected intervals, was elevated during the induction of anergy and accounted for the presence of the protein in the supernatants. In contrast, no tight coupling was observed between protein and mRNA levels for IL-2, suggesting post-translational regulation.


Evidence for a serpentinized plate interface favouring continental subduction.
The dynamics of continental subduction is largely controlled by the rheological properties of rocks involved along the subduction channel. Serpentinites have low viscosity at geological strain rates. However, compelling geophysical evidence of a serpentinite channel during continental subduction is still lacking. Here we show that anomalously low shear-wave seismic velocities are found beneath the Western Alps, along the plate interface between the European slab and the overlying Adriatic mantle. We propose that these seismic velocities indicate the stacked remnants of a weak fossilised serpentinite channel, which includes both slivers of abyssal serpentinite formed at the ocean floor and mantle-wedge serpentinite formed by fluid release from the subducting slab. Our results suggest that this serpentinized plate interface may have favoured the subduction of continental crust into the upper mantle and the formation/exhumation of ultra-high pressure metamorphic rocks, providing new constraints to develop the conceptual and quantitative understanding of continental-subduction dynamics.


Isolation and functional characterization of rat pancreatic acini.
The Authors describe an optimized procedure for the isolation of rat pancreatic acini and the preliminary results concerning the functional characterization of the cells. Isolation is carried out by two sequential digestive steps in a KREBS modified medium containing collagenase, separated by an intermediate step in which acini separation is fostered by incubation in a Ca++ free medium containing the Ca++ chelator EDTA. Final separation is obtained through the application of mechanical forces by aspirating the suspension in plastic pipettes. The choice of the collagenase, the duration and the entity of the mechanical dissociation strength are the main modifications to the classic procedure and allow to obtain a very high yield of cells maintaining their ability to secrete enzymes for a long period (6-7 hours). Functional characterization is completed mainly by the determination of the amylase release stimulated by increasing doses of cholecystokinin.


Ki-67 and Clinical Correlations in Patients with Resistant Prolactinomas.
Drug resistance is the main limitation for dopamine agonist therapy with prolactinoma. Transsphenoidal surgery is an additional operation that circumvents this limitation. To characterize distinctive clinical features of resistant prolactinoma in different levels of Ki-67, a retrospective analysis of 423 patients who were diagnosed and surgically treated at Shandong Provincial Hospital Affiliated to Shandong University from January 2007 to June 2017 was performed. Patients were subdivided into Ki-67≥3% group (High) and Ki-67<3% group (Low) according to Ki-67 labeling values. Tumors in the High group were larger (P=0.001), associated with more frequent headaches and dizziness (P=0.004), more visual impairment (P=0.008), more polyuria and polydipsia (P=0.029), and had a higher percentage of invasion (P=0.003) than patients in the Low group. However, the prolactin normalization rate in the High group was significantly lower (P=0.042). More frequent recurrence was observed in the High group (P=0.018). These observations suggest the importance of investigating Ki-67 in patients with resistant prolactinoma. The treatment of resistant prolactinomas with higher level of Ki-67 is still a challenge.


Bergmann glia function in granule cell migration during cerebellum development.
Granule cell migration influences the laminar structure of the cerebellum and thereby affects cerebellum function. Bergmann glia are derived from radial glial cells and aid in granule cell radial migration by providing a scaffold for migration and by mediating interactions between Bergmann glia and granule cells. In this review, we summarize Bergmann glia characteristics and the mechanisms underlying the effect of Bergmann glia on the radial migration of granule neurons in the cerebellum. Furthermore, we will focus our discussion on the important factors involved in glia-mediated radial migration so that we may elucidate the possible mechanistic pathways used by Bergmann glia to influence granule cell migration during cerebellum development.


Imaging in multicentric Castleman's disease.
Multicentric Castleman[']s disease (MCD) is an uncommon lymphoproliferative disorder that presents with fevers, anaemia and multifocal lymphadenopathy and nowadays is most commonly diagnosed in individuals infected with HIV-1. CT scan findings are not sufficiently reliable to establish a diagnosis of MCD which requires histological confirmation. Newer imaging techniques have therefore been studied in patients with HIV-associated MCD in an attempt to improve diagnostic accuracy. Preliminary findings suggest that FDG PET-CT may assist in the monitoring of disease activity in MCD along with familiar clinical and laboratory tools and the more recently introduced plasma HHV8 DNA viral load measurement.


Retroperitoneal Approach for Excision of Wear-Debris Pseudotumor: A Case Report.
Pelvic pseudotumors may occur as a reaction to wear-debris after hip arthroplasty and are rarely treated with surgery. We describe an instance in which a pelvic pseudotumor along the iliopsoas muscle tendon sheath was debulked using a retroperitoneal approach in a patient presenting for treatment of a prosthetic hip infection. The patient recovered uneventfully and was ambulatory with a new hip prosthesis at 3 months after procedure. Retroperitoneal exposure provided safe, excellent exposure to a wear-debris pelvic pseudotumor in this case.


Hypoxia inducible factor-1α (HIF-1α) is required for neural stem cell maintenance and vascular stability in the adult mouse SVZ.
HIF-1α is a hypoxia-inducible protein that regulates many cell and molecular processes, including those involved in angiogenesis and stem cell maintenance. Prior studies demonstrated constitutive HIF-1α stabilization in neural stem cells (NSCs) of the adult mouse SVZ, but its role there has not been elucidated. Here, we tested the hypothesis that HIF-1α plays an essential role in the maintenance of adult NSCs and stabilization of the SVZ vascular niche using conditional, tamoxifen-inducible Hif1a knock-out mice. We generated nestin-CreER(T2)/R26R-YFP/Hif1a(fl/fl) triple transgenic mice, to enable tamoxifen-inducible Hif1a gene inactivation in nestin-expressing NSCs within the adult SVZ. Hif1a gene deletion resulted in a significant loss of YFP(+) NSCs within the SVZ by 45 d post recombination, which was preceded by significant regression of the SVZ vasculature at 14 d, and concomitant decrease of VEGF expression by NSCs. Loss of YFP(+) NSCs following Hif1a gene inactivation in vivo was likely an indirect consequence of vascular regression, since YFP(+) neurosphere formation over serial passage was unaffected. These results identify NSC-encoded HIF-1α as an essential factor in the maintenance of the adult SVZ, and demonstrate that NSCs within the SVZ maintain the integrity of their vascular niche through HIF-1α-mediated signaling mechanisms.


Polyunsaturated fatty acids as antioxidants.
The susceptibility of fatty acids to oxidation is thought to be directly dependent on their degree of unsaturation. However, some in vitro and in vivo studies suggest that the relation between chemical structure and susceptibility to oxidation is not as straightforward as hypothesized from theoretical viewpoints. Indeed, long chain polyunsaturated fatty acids (LC-PUFAs) might be less oxidizable than others under specific experimental conditions. We investigated the free radical-scavenging potential of PUFA and the production of reactive oxygen/nitrogen (ROS/RNS) species by human aortic endothelial cells (HAECs) supplemented with different fatty acids. Fatty acid micelles scavenged superoxide in an unsaturation-dependent manner, up to eicosapentaenoic acid, which was the most effective fatty acid. Supplementation of HAEC with polyunsaturated fatty acids of the omega 3 series resulted in lower formation of ROS, as compared with cells supplemented with saturates, monounsaturates, or polyunsaturates of the omega 6 series. This effect was maximal at concentrations of 10muM. The effects of omega 3 fatty acids on reactive species production appear to be stronger when ROS were evaluated, as a milder, albeit significant effect was observed on RNS generation. Based on in vivo data showing reduced excretion of lipid peroxidation products after omega 3 intake and our data on ROS production and direct superoxide scavenging by LC-PUFAs, notably those of the omega 3 series, we propose that this series of fatty acid might act as indirect anti- rather than pro-oxidant in vascular endothelial cells, hence diminishing inflammation and, in turn, the risk of atherosclerosis and cardiovascular disease.


Assessing Inter-Limb Asymmetries in Soccer Players: Magnitude, Direction and Association with Performance.
In this study, we aimed to analyze the magnitude and direction of inter-limb asymmetries in ankle dorsiflexion range of motion (ROM), power (using iso-inertial devices), and a neuromuscular skill (change of direction). Secondarily, we aimed to determine the relationship between inter-limb asymmetry scores for each test and also between these scores and the scores for the different performance tests. Sixteen semiprofessional male soccer players (age: 25.38 ± 6.08 years; body height: 1.78 ± 0.64 m; body mass: 79.5 ± 14.9 kg) participated in this study. We calculated inter-limb asymmetries using five tests: ankle dorsiflexion ROM, change of direction (COD 180º), and iso-inertial resistance tasks in the open (leg extension strength (LE), leg curl strength (LC)) and closed (crossover step (CRO)) kinetic chain. Our results showed that asymmetry magnitudes differed between all tests with highest inter-limb asymmetries displayed during iso-inertial overloading. In addition, we observed that the direction of asymmetries varied depending on the test-specificity, and that the CRO asymmetries had a negative association with LE and CRO performance. These findings highlight the independent nature of asymmetries and that CRO could be an appropriate test to detect asymmetries related with the performance of soccer-specific actions (such as changes of direction). Practitioners are encouraged to use multiple tests to detect existing inter-limb differences according to the specific characteristics of each sport.


Isolation of VanB-type Enterococcus faecalis strains from nosocomial infections: first report of the isolation and identification of the pheromone-responsive plasmids pMG2200, Encoding VanB-type vancomycin resistance and a Bac41-type bacteriocin, and pMG2201, encoding erythromycin resistance and cytolysin (Hly/Bac).
Eighteen identical VanB-type Enterococcus faecalis isolates that were obtained from different hospitalized patients were examined for their drug resistance and plasmid DNAs. Of the 18 strains, 12 strains exhibited resistance to erythromycin (Em), gentamicin (Gm), kanamycin (Km), tetracycline (Tc), and vancomycin (Van) and produced cytolysin (Hly/Bac) and a bacteriocin (Bac) active against E. faecalis strains. Another six of the strains exhibited resistance to Gm, Km, Tc, and Van and produced a bacteriocin. Em and Van resistance was transferred individually to E. faecalis FA2-2 strains at a frequency of about 10(-4) per donor cell by broth mating. The Em-resistant transconjugants and the Van-resistant transconjugants harbored a 65.7-kbp plasmid and a 106-kbp plasmid, respectively. The 106-kbp and 65.7-kbp plasmids isolated from the representative E. faecalis NKH15 strains were designated pMG2200 and pMG2201, respectively. pMG2200 conferred vancomycin resistance and bacteriocin activity on the host strain and responded to the synthetic pheromone cCF10 for pCF10, while pMG2201 conferred erythromycin resistance and cytolysin activity on its host strain and responded to the synthetic pheromone cAD1 for pAD1. The complete DNA sequence of pMG2200 (106,527 bp) showed that the plasmid carried a Tn1549-like element encoding vanB2-type resistance and the Bac41-like bacteriocin genes of pheromone-responsive plasmid pYI14. The plasmid contained the regulatory region found in pheromone-responsive plasmids and encoded the genes prgX and prgQ, which are the key negative regulatory elements for plasmid pCF10. pMG2200 also encoded TraE1, a key positive regulator of plasmid pAD1, indicating that pMG2200 is a naturally occurring chimeric plasmid that has a resulting prgX-prgQ-traE1 genetic organization in the regulatory region of the pheromone response. The functional oriT region and the putative relaxase gene of pMG2200 were identified and found to differ from those of pCF10 and pAD1. The putative relaxase of pMG2200 was classified as a member of the MOB(MG) family, which is found in pheromone-independent plasmid pHTbeta of the pMG1-like plasmids. This is the first report of the isolation and characterization of a pheromone-responsive highly conjugative plasmid encoding vanB resistance.


Tobacco use among Mexican farmworkers working in tobacco: implications for agromedicine.
This survey evaluated tobacco use of migrant tobacco workers in eastern North Carolina. Sixty-nine (38%) out of 181 mostly male, Mexican farmworkers were smokers. Compared to non-smokers, three times more smokers reported alcohol use in the past week (p=0.002). More smokers compared to non-smokers reported poor to fair health, and fewer had worked previously in tobacco agriculture, but these differences were not statistically significant. Also not statistically significant, those smokers who were older and those who understood the most English smoked more cigarettes per day. Because farmworkers are exposed to many non-tobacco respiratory irritants, and because of the health risks of smoking, those who smoke should be urged to quit.


Analysis of cell lineage relationships in taste buds.
Taste buds are a heterogeneous population of cells exhibiting diverse morphological and biochemical characteristics. Because taste buds arise from multiple progenitors, the different types of taste cells may represent distinct lineages. The present study was undertaken to determine the following: (1) how many progenitors contribute to a taste bud, and (2) whether the specific subpopulation of serotonin-immunoreactive (IR) taste cells are related by lineage to a restricted set of progenitor cells. These questions were addressed using cell lineage analysis of taste buds from H253 X-inactivation mosaic mice. After random X-inactivation of the lacZ transgene, the tongue of hemizygous female mice displays discrete patches of epithelial cells, which are either beta-galactosidase (beta-gal) positive or beta-gal negative. By analyzing the proportion of the two differently stained cell populations in taste buds located at the boundary between positive and negative epithelial patches, we can determine the minimum number of progenitors that may contribute to the formation of a taste bud. The presence of taste buds containing only 6-12% labeled cells indicates that at least eight progenitors contribute to an average taste bud of 55 cells, assuming progenitors contribute equally to the cell population. Cell lineage analysis of serotonin-IR taste cells in such mixed taste buds suggests that this subpopulation likely arises from only one to two progenitors and often is related by lineage. Thus, at least some of the cell types in a taste bud represent distinct lineages of cells and are not merely phenotypic stages as a cell progresses from a young to a mature state.


Estimates of increased black carbon emissions from electrostatic precipitators during powdered activated carbon injection for mercury emissions control.
The behavior of mercury sorbents within electrostatic precipitators (ESPs) is not well-understood, despite a decade or more of full-scale testing. Recent laboratory results suggest that powdered activated carbon exhibits somewhat different collection behavior than fly ash in an ESP and particulate filters located at the outlet of ESPs have shown evidence of powdered activated carbon penetration during full-scale tests of sorbent injection for mercury emissions control. The present analysis considers a range of assumed differential ESP collection efficiencies for powdered activated carbon as compared to fly ash. Estimated emission rates of submicrometer powdered activated carbon are compared to estimated emission rates of particulate carbon on submicrometer fly ash, each corresponding to its respective collection efficiency. To the extent that any emitted powdered activated carbon exhibits size and optical characteristics similar to black carbon, such emissions could effectively constitute an increase in black carbon emissions from coal-based stationary power generation. The results reveal that even for the low injection rates associated with chemically impregnated carbons, submicrometer particulate carbon emissions can easily double if the submicrometer fraction of the native fly ash has a low carbon content. Increasing sorbent injection rates, larger collection efficiency differentials as compared to fly ash, and decreasing sorbent particle size all lead to increases in the estimated submicrometer particulate carbon emissions.


Experiences of Public Doctors on Managing Work Difficulties and Maintaining Professional Enthusiasm in Acute General Hospitals: A Qualitative Study.
Overseas studies suggest that 10-20% of doctors are depressed, 30-45% have burnout, and many report dissatisfaction with work-life balance. A local study on public doctors showed that 31.4% of the respondents satisfied the criteria for high burnout. Young, but moderately experienced doctors who need to work shifts appeared most vulnerable. This study aims to explore the experiences of those public doctors who have managed their work difficulties and maintained professional enthusiasm for references in medical education and continuing professional training. Ten public doctors with reputation were invited respectively from three acute general hospitals for an in-depth interview. Interviews were audio recorded and transcribed. Content analysis was carried out to identify major themes in relation to the research questions. Three themes emerging from difficulties encountered were (1) managing people, mostly are patients, followed by colleagues and then patients' relatives; (2) constraints at work, include time and resources; and (3) managing self with decision-making within a short time. Three themes generating from managing work difficulties included (1) self-adjustment with practicing problem solving and learning good communication appeared more frequently, followed by maintaining a professional attitude and accumulating clinical experiences; (2) seeking help from others; and (3) organizational support is also a theme though it is the least mentioned. Four themes emerging from maintaining work enthusiasm were (1) personal conviction and discipline: believing that they are helping the needy, having the sense of vocation and support from religion; disciplining oneself by continuing education, maintaining harmonious family relationship and volunteer work. (2) Challenging work: different challenging natures of their job. (3) Positive feedback from patients: positive encounters with patients keep a connectedness with their clients. (4) Organization support: working with good colleagues and opportunity for continuous training. Some implications for medical education include, developing good communication skill for medical students and junior doctors, preparing senior doctors to be mentors, and exploring the motivating force of spirituality/religion.


Arterial switch operation for transposition of the great arteries: experience from 2000-2002 in Taiwan.
Arterial switch operation (ASO) is considered the procedure of choice for transposition of great arteries (TGA). The results and long-term prognosis improved with recent advances in perioperative management. We herein analyze the clinical outcome of patients undergoing ASO at our institution during the past 3 years. From 2000 to 2002, 44 patients (30 male and 14 female) of TGA received ASO. Age at operation varied from 4 days to 6.6 years (median 14 days) with body weight ranged from 2.25 kg to 18.1 kg (median 3.3 kg). Palliative procedure prior to ASO was performed in 8 patients (18.8%). Normal coronary artery pattern was found in 28 patients (63.6%). The early mortality was 11% (5/44). Only associated ventricular septal defect (VSD) was a significant predictor for operative mortality (p=0.012). With a follow-up ranged from 11 to 44 months, the gradient of neo-pulmonary artery stenosis was 16.5 +/- 18.2 mmHg. Four patients (10%) received balloon dilatation and the other three (7.7%) underwent reoperation. The gradient of neo-aortic stenosis was 16.5 +/- 18.2 mmHg that needed to be dilated in three patients (7.7%). The probability free from reintervention was 73% at the 3rd postoperative year. One patient had moderate degree of pulmonary valve regurgitation and six had moderate neo-aortic valve regurgitation. In conclusion, the ASO can be performed in infants with satisfactory results, even in those with a body weight less than 2.5 kg. Only associated VSD was shown to be a risk factor.


Potential and emerging treatment options for Ebola virus disease.
To describe the current Ebola virus epidemic and the potential options for treatment and prevention of Ebola virus disease. A PubMed literature search (1976 through October 20, 2014) was conducted using the search term Ebola. Animal and human studies published in English were selected. Studies published within the past 5 years were the primary focus of this review. The current Ebola virus epidemic has primarily been contained in West Africa though it has subsequently spread to other areas, including the United States. The first patient in the United States infected with Ebola virus was diagnosed, treated, and expired in Texas. Two nurses caring for this patient also were diagnosed with Ebola virus and have been successfully treated. Treatment options for patients infected with Ebola virus are limited. Supportive therapy is centered on fluid resuscitation, electrolyte imbalance correction, treating complicating infections, and preventing complications of shock. Experimental therapies (ZMapp, brincidofovir, TKM-Ebola, and favipiravir) have been used during this current outbreak. Several medications such as amiodarone, chloroquine, and clomiphene may prevent the transmission of or treat Ebola virus. Different vaccine therapies are also in early-stage development. One of the vaccine strategies using recombinant vesicular stomatitis virus as a delivery vector has demonstrated efficacy when used for preexposure and postexposure prophylaxis. Ebola virus is highly virulent and fatal, and treatment options are limited. Several experimental and existing therapies may be options for preventing and treating Ebola virus disease.


[Intensive chemotherapy in myelodysplastic syndromes].
Intensive AML-like chemotherapy with anthracycline and cytosine-arabinoside or similar combinations gives lower CR rates and CR durations when administered to patients with severe MDS or AML evolving from prior MDS than to these with de novo AML. However such intensive therapies could be beneficial to younger patients without unfavorable cytogenicity when they don't have an available donor for allogeneic transplant. Numerous studies with high dose cytosine, arabinoside, idavudicine, fludarabine drugs that reverse mdr expression, or granulocytic growth factors have been conducted or are still ongoing in order to try to improve these results. In patients achieving a complete remission, intensification with autologous transplantation has also to be evaluated.


B vitamin status and concentrations of homocysteine and methylmalonic acid in elderly German women.
Prior investigations found that elderly persons are at higher risk than are younger persons for B vitamin deficiency, which leads to elevated plasma total homocysteine (tHcy) concentrations that are associated with an increased risk for certain diseases such as coronary artery disease. To date, published data have shown decreased vitamin status and elevated tHcy among the elderly. We evaluated the dietary intake and the blood status of various B vitamins and tHcy and methylmalonic acid (MMA) concentrations in 178 younger (60-70-y-old) female seniors. Dietary intake was assessed with a 3-d diet record. Thiamine, riboflavin, and vitamin B-6 activity coefficients of erythrocyte transketolase (EC 184.108.40.206), erythrocyte glutathione reductase (EC 22.214.171.124), and erythrocyte alpha-aspartic aminotransferase (EC 126.96.36.199) were used as functional indexes for the status of the 3 vitamins, respectively. Concentrations of serum and red blood cell folate, serum cobalamin and MMA, and plasma tHcy were measured. Indexes of thiamine, pyridoxine, and cobalamin indicated insufficient status in one-third of the women, whereas tHcy and MMA concentrations were elevated in 17.4% and 9.6% of the women, respectively. An association between vitamin intake and vitamin concentration in the blood was found only for folate. The mean tHcy concentration in subjects in the lowest quartile of serum folate concentration was 23% higher than that in subjects in the highest quartile. There was no association between riboflavin and tHcy concentrations. MMA was positively correlated with age and inversely correlated with serum cobalamin concentration. Even in younger, well-educated, female seniors, the prevalence of low B vitamin status and elevated plasma tHcy concentration is high. Thiamine, pyridoxine, folate, and cobalamin supplementation should be considered.


Quantifying dissociations in neuropsychological research.
Double dissociations play an important role in neuropsychology, but they are often identified through subjective estimates of "high" versus "low" performance, without considering the probability that such an outcome might have occurred by chance. To determine whether two measures "come apart" in an interesting way in brain-damaged patients, it is important to know the degree to which variance in one measure can be predicted by variance in the other. This study introduces a statistical procedure to determine the probability of a double dissociation when the correlation between measures is taken into account. Different quantitative definitions of dissociations were compared in two large samples of neurological patients, and applied to four pairs of measures (two for language, two for hemispatial neglect) with different degrees of intercorrelation (ranging from +.21 to +.84). If the correlation between measures is not taken into account, large numbers of dissociated cases may be missed, especially for measures that are highly correlated. There are also qualitative differences between methods in the identity of those individuals who meet each definition.


Clinical Investigator Training Program (CITP) - A practical and pragmatic approach to conveying clinical investigator competencies and training to busy clinicians.
Clinical investigation is a critical component of clinical medicine. Yet, other than mentorship by an experienced senior physician, young physicians have few formal training opportunities that fit into their clinical training and convey the pre-requisite clinical investigator competencies. To address this training gap, we designed the Clinical Investigator Training Program (CITP); a practical and pragmatic curriculum weaved into the constant pressures of balancing patient care with academic pursuit required of the academic practitioner. Between January 2016 and December 2018, we conducted four CITP courses, with each comprised of four 4-h sessions that included didactic lectures, group projects including the development of a mock clinical protocol, and expert's panel discussions. Each course enrolled 15 participants from an average of 28 applicants. We assessed the knowledge acquired following each course via a pre- and post-course test (t-test with positive scores indicating improvement in knowledge base). In addition, we also tracked which participants became first time principal investigator following completion of CITP. A total of 60 participants enrolled in the 4 CITP courses, and there was a statistically significant improvement in mean post-test scores (p < 0.01). The number of participants achieving first time principal investigator status nearly doubled following CITP from 17 to 33. Conversely, applicants not selected for CITP demonstrated no similar improvement during the same follow up period. The improvement in test scores and the substantive uptake in first time principal investigator responsibilities following CITP affirms that CITP provides a viable option to convey investigator competencies and encourage clinicians to take on the role of principal investigators.


Construction and expression of eukaryotic plasmids containing lamivudine-resistant or wild-type strains of Hepatitis B Virus genotype C.
To construct eukaryotic expression plasmids of full-length Hepatitis B Virus (HBV) genotype C genome, which contain lamivudine-resistant mutants (YIDD, YVDD) or wild-type strain (YMDD), and to observe the expression of HBV DNA and antigens [hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg)] of the recombinant plasmids in HepG2 cells. Three HBV full-length genomes were amplified from the plasmids pMD18T-HBV/YIDD, pMD18T-HBV/YVDD and pMD18T-HBV/YMDD, using PCR. Three recombinant plasmids were generated by inserting each of the PCR products into the eukaryotic expression vector pcDNA3.1 (+), between the EcoRI and HindIII sites. After being characterized by restriction endonuclease digestion, and DNA sequence analysis, the recombinant plasmids were transfected into HepG2 cells. At 48 and 72 h post-transfection, the levels of intracellular viral DNA replication were detected by real-time PCR, and the expression of HBsAg and HBeAg in the cell culture supernatant was determined by ELISA. Restriction endonuclease digestion and DNA sequence analysis confirmed that the three recombinant plasmids were correctly constructed. After transfecting the plasmids into HepG2 cells, high levels of intracellular viral DNA replication were observed, and HBsAg and HBeAg were secreted into the cell culture supernatant. Eukaryotic expression plasmids pcDNA3.1 (+)-HBV/YIDD, pcDNA3.1 (+)-HBV/YVDD or pcDNA3.1 (+)-HBV/YMDD, which contained HBV genotype C full-length genome, were successfully constructed. After transfection into HepG2 cells, the recombinant plasmids efficiently expressed HBV DNA, HBsAg and HBeAg. Our results provide an experimental basis for the further study of HBV lamivudine-resistant mutants.


Biphasic hypothermia in mice infected with a parasitic nematode, Trichinella spiralis.
In BALB/c mice infected with Trichinella spiralis, changes in body temperature (Tb) were observed over 35 days after the infection. T. spiralis infection induced hypothermia two times at 7 and 28 days after infection. The initial decrease persisted for about one week with a peak (37.1 +/- 0.62 degrees C) around 10 days after the infection, while the later phase persisted for at least one week. Both 10 and 35 days after the infection, there were remarkable decreases in Tb. The serum glucose level of infected mice at 10 days was significantly (p < 0.01) decreased compared with that of control mice at the same number of days, while the level in infected mice at 35 days was not decreased. Moreover, the later phase of hypothermia was prevented by the cyclooxygenase inhibitor indomethacin (10 mg/kg i.p.), while the initial phase was not. We conclude that hypothermia was caused by two different mechanisms, involving the effects of hypoglycemia and prostaglandins.


Viral IgM antibodies in serum and cerebrospinal fluid in patients with multiple sclerosis and controls.
Serum and cerebrospinal fluid (CSF) from 28 patients with multiple sclerosis (MS), 14 with other neurological diseases (OND) and 31 control subjects with tension headache were analysed for presence of IgM antibodies against measles, mumps and varicellae zoster by a specific enzyme-linked immunosorbent assay (ELISA). This technique excluded false positive reaction due to possible presence of rheumatoid factor. Twelve of the 28 patients with MS had IgM antibodies in serum and 4 in CSF, the latter always being accompanied by presence of corresponding IgM antibodies in serum. Six patients had mumps specific IgM, 5 had measles specific IgM and 3 varicellae specific IgM. In 2 patients, viral IgM antibodies were demonstrated in serum against 2 different viruses. Among the 14 OND patients, one with Wilson's disease had demonstrable serum IgM varicellae antibodies and one with radicultis had elevated serum and CSF measles and varicellae IgM antibodies. Among 31 controls, 2 had IgM antibodies in serum, one against varicellae and one against mumps. No correlations were found between viral IgM antibodies and CSF IgM index, serum IgM levels or blood-brain barrier state. Our data show that MS may be accompanied by a systemic IgM response against the 3 viruses tested, occasionally against 2 of the 3 different viruses simultaneously. The occurrence in MS of virus-specific IgM may be a reflection of viral reactivation and/or polyclonal B cell activation.