Miracle-dz/UniFAR-FTUs huggingface dataset:
An Improved Chemical Fixation Method Suitable for Immunogold Localization of Green Fluorescent Protein in the Golgi Apparatus of Tobacco Bright Yellow (BY-2) Cells

In plant systems, the green fluorescent protein (GFP) is increasingly used as a marker to study dynamics of the secretory apparatus using fluorescence microscopy. The purpose of this study was to immunogold localize the GFP, at the electron microscopic level, in a line of tobacco BY-2-cultured cells, expressing a GFP-tagged Golgi glycosyltransferase. To this end we have developed a simple, one-step chemical fixation method that allow good structural preservation and specific labeling with anti-GFP antibodies. Using this method, we have been able to show that an N-glycan GFP-tagged xylosyltransferase is specifically associated with Golgi stacks of BY-2 transformed cells and is preferentially located in medial cisternae. As an alternative to cryofixation methods, such as high-pressure freezing, which requires specialized and expensive equipment not available in most laboratories, this method offers researchers the opportunity to investigate GFP-tagged proteins of the endomembrane system in tobacco BY-2 cells.
CXCR4 receptors in the dorsal medulla: implications for autonomic dysfunction

The chemokine receptor, CXCR4, plays an essential role in guiding neural development of the CNS. Its natural agonist, CXCL12 [or stromal cell‐derived factor‐1 (SDF‐1)], normally is derived from stromal cells, but is also produced by damaged and virus‐infected neurons and glia. Pathologically, this receptor is critical to the proliferation of the HIV virus and initiation of metastatic cell growth in the brain. Anorexia, nausea and failed autonomic regulation of gastrointestinal (GI) function cause morbidity and contribute to the mortality associated with these disease states. Our previous work on the peripheral cytokine, tumor necrosis factor‐alpha, demonstrated that similar morbidity factors involving GI dysfunction are attributable to agonist action on neural circuit elements of the dorsal vagal complex (DVC) of the hindbrain. The DVC includes vagal afferent terminations in the solitary nucleus, neurons in the solitary nucleus (NST) and area postrema, and visceral efferent motor neurons in the dorsal motor nucleus (DMN) that are responsible for the neural regulation of digestive functions from the oral cavity to the transverse colon. Immunohistochemical techniques demonstrate a dense concentration of CXCR4 receptors on neurons throughout the DVC and the hypoglossal nucleus. CXCR4‐immunoreactivity is also intense on microglia within the DVC, though not on the astrocytes. Physiological studies show that nanoinjection of SDF‐1 into the DVC produces a significant reduction in gastric motility in parallel with an elevation in the numbers of cFOS‐activated neurons in the NST and DMN. These results suggest that this chemokine receptor may contribute to autonomically mediated pathophysiological events associated with CNS metastasis and infection.
THE 2007 OTTO AUFRANC AWARD: Ceramic-on-Metal Hip Arthroplasties: A Comparative In Vitro and In Vivo Study

The performance of novel ceramic-on-metal bearing couples has been compared with metal-on-metal and ceramic-on-ceramic bearing couples in laboratory and short-term clinical studies. Laboratory studies compared ceramic-on-metal with metal-on-metal and ceramic-on-ceramic bearings with diameters of 28 and 36 mm under standard conditions and under adverse conditions with head loading on the rim of the cup. Clinical studies compared metal ion levels in ceramic-on-metal with metal-on-metal, ceramic-on-ceramic, and ceramic-on-polyethylene bearings in a randomized prospective study. In the laboratory studies, friction, wear, and ion levels were lower in ceramic-on-metal bearings compared with metal-on-metal, with results similar to ceramic-on-ceramic couples. Under adverse conditions and rim loading, all bearings showed increased wear with lower wear and absence of stripe wear in ceramic-on-metal compared with metal-on-metal bearings. Short-term studies in 31 patients at 6 months revealed lower metal ion levels (cobalt and chromium) in those with ceramic-on-metal compared with metal-on-metal bearings.Level of Evidence: Level II, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence.
Motor cortex excitability and fatigue in multiple sclerosis: a transcranial magnetic stimulation study

We investigated electrophysiological correlates of fatigue in patients with multiple sclerosis (MS). Transcranial magnetic stimulation (TMS) was used to explore motor excitability in three groups of subjects: MS patients with fatigue (MS-F), MS patients without fatigue (MS-NF) and healthy control subjects. All participants had to perform a fatiguing hand-grip exercise. TMS was performed prior to and after the exercise. Prior to the motor task, MS-F patients had less inhibition in the primary motor cortex compared to both other groups. Postexercise, intracortical inhibition was still reduced in the MS-F patients compared to the MS-NF patients. In MS-F patients the postexercise time interval for normalization of the motor threshold was correlated with the fatigue severity. We conclude that MS patients with fatigue have an impairment of inhibitory circuits in their primary motor cortex. The results also indicate that fatigue severity is associated with an exercise-induced reduction of membrane excitability.
Palivizumab for the prevention of respiratory syncytial virus infection.

QUESTION Palivizumab, a specific monoclonal antibody for respiratory syncytial virus (RSV), is available for prevention of pediatric respiratory tract infections. What are the indications for its use and can it be used for treatment of RSV infections? ANSWER Most infants should not be considered for RSV prophylaxis with palivizumab. The drug is approved for use for different indications in different Canadian provinces. The drug should be administered only in the context of infants most vulnerable to severe RSV illness with a high likelihood of hospital admission, particularly in the first 6 months of life. It is not effective in the treatment of RSV disease and it is not approved or recommended for this indication.
Satellite DNA Modulates Gene Expression in the Beetle Tribolium castaneum after Heat Stress

Non-coding repetitive DNAs have been proposed to perform a gene regulatory role, however for tandemly repeated satellite DNA no such role was defined until now. Here we provide the first evidence for a role of satellite DNA in the modulation of gene expression under specific environmental conditions. The major satellite DNA TCAST1 in the beetle Tribolium castaneum is preferentially located within pericentromeric heterochromatin but is also dispersed as single repeats or short arrays in the vicinity of protein-coding genes within euchromatin. Our results show enhanced suppression of activity of TCAST1-associated genes and slower recovery of their activity after long-term heat stress relative to the same genes without associated TCAST1 satellite DNA elements. The level of gene suppression is not influenced by the distance of TCAST1 elements from the associated genes up to 40 kb from the genes’ transcription start sites, but it does depend on the copy number of TCAST1 repeats within an element, being stronger for the higher number of copies. The enhanced gene suppression correlates with the enrichment of the repressive histone marks H3K9me2/3 at dispersed TCAST1 elements and their flanking regions as well as with increased expression of TCAST1 satellite DNA. The results reveal transient, RNAi based heterochromatin formation at dispersed TCAST1 repeats and their proximal regions as a mechanism responsible for enhanced silencing of TCAST1-associated genes. Differences in the pattern of distribution of TCAST1 elements contribute to gene expression diversity among T. castaneum strains after long-term heat stress and might have an impact on adaptation to different environmental conditions.
Paternal transcripts for glucose‐6‐phosphate dehydrogenase and adenosine deaminase are first detectable in the human preimplantation embryo at the Three‐ to Four‐Cell stage

The transition between dependence on maternal transcripts and proteins inherited in the oocyte and embryonic gene expression in the human preimplantation embryo occurs at the four‐ to eight‐cell stage. Recently, studies using reverse transcriptase polymerase chain reaction (RT‐PCR) have detected paternal transcripts for the Y‐linked genes, ZFY and SRY, and the myotonic dystrophy associated protein kinase gene, DK, as early as the late pronucleate one‐cell stage. However, expression at the protein level has not been demonstrated and its function at these early stages is unknown. Using coding sequence polymorphisms to distinguish maternal and paternal transcripts, we have examined the transcription of two ubiquitously expressed genes: X‐linked glucose‐6‐phosphate dehydrogenase (G6PD) and adenosine deaminase (ADA). Both G6PD and ADA are housekeeping genes with TATA‐less promoters which, because of their roles in metabolism and ubiquitous expression, may provide a more reliable indication of the timing of activation of the embryonic genome. They also each have biallelic polymorphisms with a high heterozygosity ratio which can be detected by restriction digestion. Couples undergoing in vitro fertilization (IVF) were screened for these polymorphisms. Individual spare oocytes and embryos at different stages of preimplantation development were analyzed by RT‐PCR and appropriate restriction digestion in those cases in which the male partner carried a different allele to the female partner. In addition, since only female embryos inherit the paternal allele of X‐linked G6PD, cDNA was also analyzed for ZFX/ZFY transcripts to identify the sex of each embryo. One hundred and twenty three individual oocytes and embryos were analyzed by RT‐PCR and restriction digestion to detect the paternal transcripts from the polymorphic alleles. Maternal transcripts for G6PD, ADA, and ZFX were detected in all oocytes and embryos and at all stages.
Hemorrhagic and thrombotic complications in patients with myeloproliferative diseases.

OBJECTIVE To correlate the incidence of hemorrhage and thrombosis to bleeding time (BT) and platelet aggregation in 27 consecutive patients with myeloproliferative diseases (MPD). DESIGN Retrospective study. SETTING Public tertiary referral center. PATIENTS Eighteen patients with chronic myelogenous leukemia (CML), 5 with polycytemia vera (PV), 2 with essential thrombocytemia (ET) and 2 with idiopathic myelofibrosis (MF). Duke's BT and epinephrine-induced platelet aggregation were performed on the patients and on 10 healthy individuals. RESULTS Eleven patients presented symptoms (41%):9 with hemorrhage (33%) and 5 with thrombosis (19%). There were less symptomatic patients in the CML group (28%) than in the other MPD (67%), without statistical significance (Fisher, p = 0.06). Duke's BT was longer in symptomatic patients (Mann-Whitney, p < 0.05). Platelet aggregation was abnormal in 7 patients (26%) and 71% of them were symptomatic (Fisher, p = 0.07). CONCLUSIONS The high incidence of bleeding and thrombosis in patients with MPD was related to prolonged BT, but not to platelet aggregation abnormalities.
Radiographic analysis of the sagittal alignment and balance of the spine in asymptomatic subjects.

BACKGROUND There is an increasing recognition of the clinical importance of the sagittal plane alignment of the spine. A prospective study of several radiographic parameters of the sagittal profile of the spine was conducted to determine the physiological values of these parameters, to calculate the variations of these parameters according to epidemiological and morphological data, and to study the relationships among all of these parameters. METHODS Sagittal radiographs of the head, spine, and pelvis of 300 asymptomatic volunteers, made with the subject standing, were evaluated. The following parameters were measured: lumbar lordosis, thoracic kyphosis, T9 sagittal offset, sacral slope, pelvic incidence, pelvic tilt, intervertebral angulation, and vertebral wedging angle from T9 to S1. The radiographs were digitized, and all measurements were performed with use of a software program. Two different analyses, a descriptive analysis characterizing these parameters and a multivariate analysis, were performed in order to study the relationships among all of them. RESULTS The mean values (and standard deviations) were 60 degrees 10 degrees for maximum lumbar lordosis, 41 degrees +/- 8.4 degrees for sacral slope, 13 degrees +/- 6 degrees for pelvic tilt, 55 degrees +/-10.6 degrees for pelvic incidence, and 10.3 degrees +/- 3.1 degrees for T9 sagittal offset. A strong correlation was found between the sacral slope and the pelvic incidence (r = 0.8); between maximum lumbar lordosis and sacral slope (r = 0.86); between pelvic incidence and pelvic tilt (r = 0.66); between maximum lumbar lordosis and pelvic incidence, pelvic tilt, and maximum thoracic kyphosis (r = 0.9); and, finally, between pelvic incidence and T9 sagittal offset, sacral slope, pelvic tilt, maximum lumbar lordosis, and thoracic kyphosis (r = 0.98). The T9 sagittal offset, reflecting the sagittal balance of the spine, was dependent on three separate factors: a linear combination of the pelvic incidence, maximum lumbar lordosis, and sacral slope; the pelvic tilt; and the thoracic kyphosis. CONCLUSIONS AND CLINICAL RELEVANCE This description of the physiological spinal sagittal balance should serve as a baseline in the evaluation of pathological conditions associated with abnormal angular parameter values. Before a patient with spinal sagittal imbalance is treated, the reciprocal balance between various spinal angular parameters needs to be taken into account. The correlations between angular parameters may also be useful in calculating the corrections to be obtained during treatment.
Functional flexibility and acclimation of the thylakoid membrane

Light is an elusive substrate for the function of photosynthetic light reactions of photosynthesis in the thylakoid membrane. Therefore structural and functional dynamics, which occur in the timescale from seconds to several days, are required both at low and high light conditions. The best characterized short-time regulation mechanism at low light is a rapid state transition, resulting in higher absorption cross section of PSI at the expense of PSII. If the low light conditions continue, activation of the lhcb -genes and synthesis of the light-harvesting proteins will occur to optimize the functions of PSII and PSI. At high light, the transition to state 2 is completely inhibited, but the feedback de-excitation of absorbed energy as heat, known as the energy-dependent quenching (q_E), is rapidly set up. It requires, at least, the ΔpH-dependent activation of violaxanthin de-epoxidase and involvement of the PsbS protein. Another crucial mechanism for protection against the high light stress is the PSII repair cycle. Furthermore, the water-water cycle, cyclic electron transfer around PSI and chlororespiration are important means induced under high irradiation, functioning mainly to avoid an excess production of reactive oxygen species.
Functional outcome of patients undergoing replantation of hand at wrist level-7 year experience

Background: Replantation is defined as reattachment of amputated limb using neurovascular and musculoskeletal structures in order to obtain recovery of limb. Re-vascularisation involves all the above steps in case of limb injuries that result in a near total amputation. Aim and Objective: To study the functional outcome of patients undergoing replantation of hand at wrist level. Material and Methods: This is a retrospective study of patients who underwent replantation of total amputation of hand at wrist level within a period of Jan 2003-June 2010. We evaluated post operative functional outcome compared to uninjured hand taking into consideration: 1. The patient's overall satisfaction with the hand. 2. Recovery of flexor and extensor function of thumb and fingers. 3. Recovery of thumb opposition. 4. Recovery of sensations in the median and ulnar nerve distribution. 5. Ability of surviving hand to perform daily tasks. Results: There were total seventeen patients and age range was two years to 55 years. Out of 17 patients,16 were males. All the replantations were successful except for one. Summary: The results showed that, although the replanted hands were never functionally as good as the contralateral hand the patients were able to perform most of the daily activities.
Titanium-nitride-oxide-coated coronary stents: insights from the available evidence

Abstract Coating of stent surface with a biocompatible material is suggested to improve stent safety profile. A proprietary process was developed to coat titanium-nitride-oxide on the stent surface, based on plasma technology that uses the nano-synthesis of gas and metal. Preclinical in vitro and in vivo investigation confirmed blood compatibility of titanium (nitride-) oxide films. Titanium-nitride-oxide-coated stents demonstrated a better angiographic outcome, compared with bare-metal stents at mid-term follow-up; however, they failed to achieve non-inferiority for angiographic outcome versus second-generation drug-eluting stents. Observational studies showed adequate clinical outcome at mid-term follow-up. Non-randomized studies showed an outcome of titanium-nitride-oxide-coated stents comparable to – or better than – first-generation drug-eluting stents at long-term follow-up. Two randomized controlled trials demonstrated comparable efficacy outcome, and a better safety outcome of titanium-nitride-oxide-coated stents versus drug-eluting stents at long-term follow-up. Evaluation by optical coherence tomography at mid-term follow-up revealed better neointimal strut coverage associated with titanium-nitride-oxide-coated stents versus drug-eluting stents; yet, neointimal hyperplasia thickness was greater. Key messages Stents coated with titanium-nitride-oxide demonstrated biocompatibility in preclinical studies: they inhibit platelet and fibrin deposition, and reduce neointimal growth. In observational and non-randomized studies, titanium-nitride-oxide-coated stents were associated with adequate safety and efficacy outcome. In randomized trials of patients with acute coronary syndrome, titanium-nitride-oxide-coated stents were associated with a better safety outcome, compared with drug-eluting stents; efficacy outcome was comparable.
Microbial Ecology of the Dark Ocean above, at, and below the Seafloor

SUMMARY The majority of life on Earth—notably, microbial life—occurs in places that do not receive sunlight, with the habitats of the oceans being the largest of these reservoirs. Sunlight penetrates only a few tens to hundreds of meters into the ocean, resulting in large-scale microbial ecosystems that function in the dark. Our knowledge of microbial processes in the dark ocean—the aphotic pelagic ocean, sediments, oceanic crust, hydrothermal vents, etc.—has increased substantially in recent decades. Studies that try to decipher the activity of microorganisms in the dark ocean, where we cannot easily observe them, are yielding paradigm-shifting discoveries that are fundamentally changing our understanding of the role of the dark ocean in the global Earth system and its biogeochemical cycles. New generations of researchers and experimental tools have emerged, in the last decade in particular, owing to dedicated research programs to explore the dark ocean biosphere. This review focuses on our current understanding of microbiology in the dark ocean, outlining salient features of various habitats and discussing known and still unexplored types of microbial metabolism and their consequences in global biogeochemical cycling. We also focus on patterns of microbial diversity in the dark ocean and on processes and communities that are characteristic of the different habitats.
Species richness and susceptibility to heat and drought extremes in synthesized grassland ecosystems: compositional vs physiological effects

Summary 1 We investigated effects of declining plant species richness (S) on resistance to extremes in grassland communities. 2 Synthesized model ecosystems of different S, grown outdoors in containers, were exposed to a stress peak combining heat and drought. The heat wave was induced experimentally by infrared irradiation in free air conditions. 3 Before the heat wave, the more species-rich communities produced more biomass as a result of a large and positive complementarity effect that outweighed a small negative selection effect. 4 Water use during the heat wave was likewise enhanced by S, which could not be attributed to dominance of ‘water-wasting’ species. Instead, water consumption at high S exceeded that expected from changes in community biomass and biomass composition. The observed enhancement of resource (water) acquisition under stress with increasing S therefore probably originated from complementarity. 5 Despite enhanced water use in the more diverse communities, plant survival was significantly less, affecting all species alike. Physiological stress, recorded as photochemical efficiency of photosystem II electron transport, was significantly greater. Before the heat wave, the changes in biomass composition that coincided with increasing S did not favour species that would later prove intrinsically sensitive or insensitive. 6 Complementarity in resource use for biomass production had a cost in terms of reduced survival under stress, despite the likelihood of complementarity in water acquisition during exposure. The greater loss of individuals from the more diverse grasslands suggests enhanced risk of local extinction.
TOXIN DETECTION IN Pasteurella multocida STRAINS ISOLATED FROM SWINE LUNGS IN THE STATE OF RIO GRANDE DO SUL , BRAZIL

Toxin production and the presence of the toxA gene was assayed in 24 Pasteurella multocida strains isolated from pigs with pneumonia and pleuritis in the State of Rio Grande do Sul, Brazil. Toxigenicity was assessed through a mouse lethality test and a PCR amplification assay was used to obtain a 300 bp product from the toxA gene. In order to demonstrate that the amplified product corresponded to the toxA gene, the same segment of a reference P. multocida capsular type D toxigenic strain was amplified and treated with the restriction enzyme Taq1, and the restriction fragments corresponded to the expected sizes. Only one out of 24 strains of P. multocida was toxigenic in the mouse lethality test. However, with the use of PCR, nine strains carried the toxA gene. Hypothesis to explain this difference is the suppression of genic expression in vivo or the presence of a defective, non-functional gene.
Microvascular retinopathy in subjects without diabetes: the Inter99 Eye Study

Purpose: Retinal vascular lesions such as microaneurysms and haemorrhages, while typical of diabetic retinopathy, are also seen in subjects without diabetes where they are associated with elevated cardiovascular mortality. In theory, these lesions could be a consequence of past hyperglycaemia. We examined the prevalence and risk factors for retinopathy, including lens fluorescence, a biomarker of cumulative life‐time glycaemia in adults without diabetes.