Miracle-dz/UniFAR-FTUs huggingface dataset:
Proton Transfers in a Channelrhodopsin-1 Studied by Fourier Transform Infrared (FTIR) Difference Spectroscopy and Site-directed Mutagenesis*

Background: Channelrhodopsin-1 is a red-shifted light-gated cation channel. Results: Proton transfers and H-bonding changes involving the Schiff base counterion residues Asp-169 and Glu-299 were detected. Conclusion: A two-step proton transfer relay mechanism consistent with our results and earlier photoinduced channel current and pH titration measurements is proposed. Significance: Understanding the mechanism of channelrhodopsin can lead to improved optogenetic control of neurons. Channelrhodopsin-1 from the alga Chlamydomonas augustae (CaChR1) is a low-efficiency light-activated cation channel that exhibits properties useful for optogenetic applications such as a slow light inactivation and a red-shifted visible absorption maximum as compared with the more extensively studied channelrhodopsin-2 from Chlamydomonas reinhardtii (CrChR2). Previously, both resonance Raman and low-temperature FTIR difference spectroscopy revealed that unlike CrChR2, CaChR1 under our conditions exhibits an almost pure all-trans retinal composition in the unphotolyzed ground state and undergoes an all-trans to 13-cis isomerization during the primary phototransition typical of other microbial rhodopsins such as bacteriorhodopsin (BR). Here, we apply static and rapid-scan FTIR difference spectroscopy along with site-directed mutagenesis to characterize the proton transfer events occurring upon the formation of the long-lived conducting P2380 state of CaChR1. Assignment of carboxylic C=O stretch bands indicates that Asp-299 (homolog to Asp-212 in BR) becomes protonated and Asp-169 (homolog to Asp-85 in BR) undergoes a net change in hydrogen bonding relative to the unphotolyzed ground state of CaChR1. These data along with earlier FTIR measurements on the CaChR1 → P1 transition are consistent with a two-step proton relay mechanism that transfers a proton from Glu-169 to Asp-299 during the primary phototransition and from the Schiff base to Glu-169 during P2380 formation. The unusual charge neutrality of both Schiff base counterions in the P2380 conducting state suggests that these residues may function as part of a cation selective filter in the open channel state of CaChR1 as well as other low-efficiency ChRs.
Gene therapy for HIV/AIDS: the potential for a new therapeutic regimen

Human Immunodeficiency Virus (HIV) is the etiologic agent of Acquired Immunodeficiency Syndrome (AIDS). HIV/AIDS is a disease that, compared with the not so distant past, is now better held in check by current antiretroviral drugs. However, it remains a disease not solved. Highly active antiretroviral therapy (HAART) generally uses two non‐nucleoside and one nucleoside reverse transcriptase (RT) inhibitor or two non‐nucleoside RT and one protease inhibitor. HAART is far more effective than the mono‐ or duo‐therapy of the past, which used compounds like the nucleoside reverse transcriptase inhibitor AZT or two nucleoside reverse transcriptase inhibitors. However, even with the relatively potent drug cocktails that comprise HAART, there are the issues of (i) HIV escape mutants, (ii) an apparent need to take the drugs in an ongoing manner, and (iii) the drugs' side effects that are often severe. This review speaks to the potential addition to these potent regimens of another regimen, namely the genetic modification of target hematopoietic cells. Such a new treatment paradigm is conceptually attractive as it may yield the constant intracellular expression of an anti‐HIV gene that acts to inhibit HIV replication and pathogenicity. A body of preclinical work exists showing the inhibition of HIV replication and decreased HIV pathogenicity by anti‐HIV genetic agents. This preclinical work used hematopoietic cell lines and primary cells as the target tissue. More recently, several clinical trials have sought to test this concept in vivo. Copyright © 2003 John Wiley & Sons, Ltd.
Temporary transvenous pacing: endangered skill

Background Temporary cardiac pacing although is an essential requirement for core medical training (CMT) in UK, there are no defined training measures and guidelines available as to who should perform this. Methods We conducted an anonymous survey of 300 non-cardiology medical registrars regarding their individual ability, experience and training received in temporary pacing wire (TPW) insertion. Results A total of 202 (67%) responses were received. 61% (123) had not performed any TPW insertion before becoming a registrar. Only18% (38) felt confident in inserting a TPW unsupervised and only14 (7%) had ever received any formal training. The majority, 169 (84%), did not feel that their on-call consultant general physician would be able to perform the procedure. Conclusion This survey shows that general medical registrars lack a major life-saving skill that is required as part of CMT. Thus, there is now an urgent clinical governance need to either formally train physicians or abandon the concept and practice of general internal medicine-led temporary pacing, and devolve this to cardiologists.
Leptin Protects the Pancreas from Damage Induced by Caerulein Overstimulation by Modulating Cytokine Production

Background: Recent identification of specific leptin receptors in the pancreas suggests that this peptide may also play some role in this gland. Aim: To examine the effect of intraperitoneal (i.p.) or intracerebroventricular (i.c.v.) administration of leptin in rats on caerulein-induced pancreatitis (CIP), pancreatic gene expression of leptin and inflammatory cytokine production. Methods: caerulein (25 µg/kg) was infused subcutaneously into conscious rats over 5 h to produce CIP. Leptin (1, 5, or 10 µg/kg) was injected i.p. or i.c. v. 30 min prior to the CIP induction. The plasma level of TNFα and IL-4 was determined by ELISA, while plasma leptin was measured by RIA and leptin gene expression in pancreas by RT-PCR. Results: CIP was characterized by the usual pancreatic edema, reduction in pancreatic blood flow (PBF) and an increase in serum levels of amylase, TNFα and IL-4. Pretreatment with i.p. or i.c. v. leptin of the CIP rats partially reversed the harmful effects of CIP on the pancreas, and reduced pancreatic inflammation and the fall in PBF. This was accompanied by a dose-dependent reduction in serum levels of amylase and TNFα, while serum IL-4 in the CIP rats pretreated with leptin rose dose-dependently as compared to control rats with CIP alone. Pretreatment with leptin resulted in the dose-dependent rise in plasma leptin level over that observed in vehicle-treated controls. Leptin mRNA expression in the pancreas was dose-dependently increased after infusion of caerulein. Leptin content in isolated pancreatic acini was also increased dose-dependently by caerulein added to the incubation medium bathing these acini. Conclusions: (1) Exogenous leptin protects the pancreas against damage by CIP; (2) endogenous leptin seems to limit the extend of pancreatic damage, and (3) these protective effects of leptin could be attributed to the reduction in TNFα and to the increase in IL-4 production.
ELEVATED EPIDERMAL GROWTH FACTOR RECEPTOR LEVELS IN HYPERTENSIVE LYON RAT KIDNEY AND AORTA

1. The adult spontaneously hypertensive Lyon rats (LH strain) exhibited increased maximal epidermal growth factor (EGF) binding in freshly prepared kidney and aortic tissue membranes compared with age‐matched normotensive (LN) or hypotensive (LL) strains. However, the binding affinity of the receptors to EGF was the same in all the three strains studied. These findings indicate an increased number of EGF receptors (EGFR) in the hypertensive LH strain.
Synaptic organization of cortico‐cortical connections from the primary visual cortex to the posteromedial lateral suprasylvian visual area in the cat

The synaptic organization of the projection from the cat striate visual cortex to the posteromedial lateral suprasylvian cortical area (PMLS) was examined. The anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA‐L) was iontophoretically delivered into area 17, and anterogradely labeled fibers were revealed in PMLS by means of an immunocytochemical detection method. Most axons and presumptive terminal swellings were found in layers III and IV. The neuronal elements (n = 190) that were postsynaptic to anterogradely labeled boutons were quantitatively analyzed. All anterogradely labeled cortico‐cortical boutons (n = 182) established type 1 synapses. The results show that 83% of the postsynaptic targets were dendritic spines, probably belonging to pyramidal cells. Dendritic shafts constituted 17% of the targets. The dendritic shafts postsynaptic to cortico‐cortical boutons were studied for the presence of gamma‐aminobutyric acid (GABA) with a postembedding immunogold method. Most dendritic shafts (85%) that were tested were found to be GABA‐positive, demonstrating that they originate from local inhibitory neurons. Taking into account that most postsynaptic targets were spines and extending the results of the immunocytochemical testing to the total population of postsynaptic dendrites, it was calculated that at least 14% of targets originated from GABA‐positive cell.
Facilitation of currents through rat Ca2+‐permeable AMPA receptor channels by activity‐dependent relief from polyamine block

1 In outside‐out patches excised from human embryonic kidney (HEK) 293 cells expressing Ca2+‐permeable α‐amino‐3‐hydroxy‐5‐methyl‐4‐isoxazole‐propionate receptor (AMPAR) channels, currents activated by 1 ms glutamate pulses at negative membrane potentials facilitated during and following a repetitive (2 to 100 Hz) agonist application. The degree of facilitation depended on subunit type, membrane potential and stimulation frequency being antagonized by a slow recovery from desensitization. 2 Activity‐dependent current facilitation occurred in Ca2+‐permeable but not in Ca2+‐impermeable AMPAR channels. Current facilitation, however, does not depend on Ca2+ flux. Rather it reflects a relief from the block of Ca2+‐permeable AMPARs by intracellular polyamines since facilitation occurred only in the presence of polyamines and since facilitated currents had a nearly linear current‐voltage relation (I‐V). 3 Relief from polyamine block was use dependent and occurred mainly in open channels. The relief mechanism was determined primarily by membrane potential rather than by current flow. 4 In closed channels the degree of polyamine block was independent of membrane potential. The voltage dependence of the rate of relief from the block in open channels rather than the voltage dependence of the block underlies the inwardly rectifying shape of the I‐V at negative potentials. 5 Currents through native Ca2+‐permeable AMPAR channels in outside‐out or nucleated patches from either hippocampal basket cells or a subtype of neocortical layer II nonpyramidal cells also showed facilitation. 6 It is concluded that a use‐dependent relief from polyamine block during consecutive AMPAR channel openings underlies current facilitation. This polyamine‐AMPAR interaction may represent a new activity‐dependent postsynaptic mechanism for control of synaptic signalling.
The Pliocene Paradox (Mechanisms for a Permanent El Niño)

During the early Pliocene, 5 to 3 million years ago, globally averaged temperatures were substantially higher than they are today, even though the external factors that determine climate were essentially the same. In the tropics, El Niño was continual (or “permanent”) rather than intermittent. The appearance of northern continental glaciers, and of cold surface waters in oceanic upwelling zones in low latitudes (both coastal and equatorial), signaled the termination of those warm climate conditions and the end of permanent El Niño. This led to the amplification of obliquity (but not precession) cycles in equatorial sea surface temperatures and in global ice volume, with the former leading the latter by several thousand years. A possible explanation is that the gradual shoaling of the oceanic thermocline reached a threshold around 3 million years ago, when the winds started bringing cold waters to the surface in low latitudes. This introduced feedbacks involving ocean-atmosphere interactions that, along with ice-albedo feedbacks, amplified obliquity cycles. A future melting of glaciers, changes in the hydrological cycle, and a deepening of the thermocline could restore the warm conditions of the early Pliocene.
Effect of Ethanol, Acetaldehyde, and Acetate on Insulin and Glucagon Secretion in the Perfused Rat Pancreas

The effects of varying concentrations of ethanol (1,10, and 30 mM) and its metabolites (1 mM acetate and 1 and 10 mM acetaldehyde) on insulin and glucagon secretion induced by glucose (11.1 mM) and arginine (20 mM) were studied in isolated perfused pancreas of Sprague-Dawley rats Ethanol and its metabolites did not significantly modify basal secretion of the two hormones. Ethanol reduced glucose-induced insulin secretion by means of a dose-related effect. Arginine-induced insulin output did not seem to be influenced to any significant degree. Acetate and acetaldehyde significantly inhibited glucose and arginine-induced insulin secretion. While ethanol (10 and 30 mM) did not modify glucagon output during arginine perfusion, acetate and acetaldehyde markedly enhanced it. The block of insulin secretion and the increased secretion of glucagon could explain the diabetogenic effect of ethanol demonstrated in vivo. The mechanism by which ethanol acts on the pancreatic β- and α-cells is discussed.
My choice, your problem? Mandating IT use in large organisational networks

On the basis of a four‐year exploratory study of a mandatory information systems implementation by an Italian, multibillion‐dollar dairy cooperative with 2200 members, this paper describes how key stakeholders engage in dynamic transformation processes that shape the technology, the users' practices and the organisation itself. In doing so, this study responds to calls for process‐oriented longitudinal explorations and suggests an alternative path of adoption in which the technology becomes the reification of a bi‐directional discourse about the transformation of practices for the entire network of organisations. In presenting this alternative path, this study unveils a five‐phase change process that both altered perceptions of the technology and its possibilities at the same time resolving tensions among the drivers and users of the mandated system. © 2016 John Wiley & Sons Ltd
Evaluation of the sedative and cardiovascular effects of intramuscular administration of dexmedetomidine with and without concurrent atropine administration in dogs.

OBJECTIVE To evaluate degree of sedation and cardiovascular, respiratory, acid-base excess, and electrolyte variables in response to IM administration of dexmedetomidine or dexmedetomidine with atropine. DESIGN Randomized crossover study. Animals-5 healthy 1- to 2-year-old sexually intact male Treeing Walker Coonhounds. PROCEDURES Dogs were instrumented with catheters placed in the dorsal pedal artery and lateral saphenous vein. All dogs received dexmedetomidine (10 μg/kg [4.5 μg/lb], IM) or dexmedetomidine with atropine (0.02 mg/kg [0.009 mg/lb], IM). Variables were measured at baseline (time 0) and 5, 15, 30, and 60 minutes after drug administration. RESULTS In all dogs, lithium dilution cardiac output decreased from a mean ± SD baseline value of 5.07 ± 1.0 L/min to 2.1 ± 0.9 L/min. Cardiac output was not different between dexmedetomidine group dogs and dexmedetomidine-atropine group dogs. Mean arterial pressure increased from baseline in both groups but was significantly higher in dexmedetomidine-atropine group dogs, compared with dexmedetomidine group dogs. Heart rate in dexmedetomidine group dogs decreased from 110 ± 14.2 beats/min to 49.4 ± 10.4 beats/min by 15 minutes. No differences were seen in blood gas values, electrolyte concentration, or hemoglobin values over time or between groups. Arrhythmias were detected in dexmedetomidine-atropine group dogs and included atrioventricular block, ventricular premature contractions, and ventricular bigeminy. CONCLUSIONS AND CLINICAL RELEVANCE Administration of atropine at 0.02 mg/kg, IM, with dexmedetomidine at 10 μg/kg, IM, resulted in an increase in mean arterial blood pressure and heart rate; deleterious cardiac arrhythmias were also observed. Use of atropine with dexmedetomidine is not recommended in dogs.
Capsid Functions of Inactivated Human Picornaviruses and Feline Calicivirus

ABSTRACT The exceptional stability of enteric viruses probably resides in their capsids. The capsid functions of inactivated human picornaviruses and feline calicivirus (FCV) were determined. Viruses were inactivated by UV, hypochlorite, high temperature (72°C), and physiological temperature (37°C), all of which are pertinent to transmission via food and water. Poliovirus (PV) and hepatitis A virus (HAV) are transmissible via water and food, and FCV is the best available surrogate for the Norwalk-like viruses, which are leading causes of food-borne and waterborne disease in the United States. The capsids of all 37°C-inactivated viruses still protected the viral RNA against RNase, even in the presence of proteinase K, which contrasted with findings with viruses inactivated at 72°C. The loss of ability of the virus to attach to homologous cell receptors was universal, regardless of virus type and inactivation method, except for UV-inactivated HAV, and so virus inactivation was almost always accompanied by the loss of virus attachment. Inactivated HAV and FCV were captured by homologous antibodies. However, inactivated PV type 1 (PV-1) was not captured by homologous antibody and 37°C-inactivated PV-1 was only partially captured. The epitopes on the capsids of HAV and FCV are evidently discrete from the receptor attachment sites, unlike those of PV-1. These findings indicate that the primary target of UV, hypochlorite, and 72°C inactivation is the capsid and that the target of thermal inactivation (37°C versus 72°C) is temperature dependent.
Neutropenia and febrile neutropenia in patients with Hodgkin's lymphoma treated with doxorubicin (Adriamycin), bleomycin, vinblastine and dacarbazine (ABVD) chemotherapy

When uncomplicated neutropenia during doxorubicin (Adriamycin), bleomycin, vinblastine and dacarbazine (ABVD) chemoterapy for the treatment of Hodgkin's lymphoma is encountered, it is unclear whether or not treatment should be modified. In the present study, we determined the incidence of neutropenia, febrile neutropenia, and the relationship of febrile neutropenia to grade III/IV neutropenia and dose modification, in a large university patient population. We reviewed the charts of patients diagnosed with Hodgkin's lymphoma between 1 January 1990 and 31 December 2002 who were treated with ABVD chemotherapy, and seen at the University of Iowa with complete diagnosis, staging, and treatment dosing records. Adequate data was available on 894 treatments in 81 patients with Hodgkin's lymphoma treated with ABVD chemotherapy. Grade III/IV neutropenia was present on the scheduled day of treatment in 187 (20.9%) treatments in 64 (79%) patients. Grade III/IV neutropenia was most common at cycle 1 day 15. Febrile neutropenia developed nine times in eight patients, and eight episodes of febrile neutropenia developed when the treatment-day absolute neutrophil count (ANC) ⩾1000. Dose delay of >4 days and/or dose reduction to <80% of original doxorubicin dose following grade III/IV neutropenia occurred in 29 of 187 treatments, with no episodes of febrile neutropenia. With grade III/IV neutropenia on the day of therapy, 158 treatments were administered without dose reduction or dose delay with one subsequent episode of febrile neutropenia. Neutropenia during ABVD is common, and dose modification for uncomplicated neutropenia on the day of treatment may not reduce the risk of febrile neutropenia. It may be possible to maintain dose intensity in the face of uncomplicated neutropenia during ABVD therapy.
Hyperbolic Heat Transfer Equation for Radiofrequency Heating: Comparison between Analytical and COMSOL solutions

The radiofrequency heating (RFH) is widely employed to heat biological tissue in different surgical procedures. Most models analyze the RFH employing a parabolic heat transfer equation (PHTE) based on Fourier’s theory. The PHTE can be used for problems involving long heating times or low thermal gradients. However, when the problem involves short heating times or extreme thermal gradients it is needed to solve it using the hyperbolic heat transfer equation (HHTE), where a wavy behavior of heat conduction with a finite thermal propagation speed is assumed . We have recently proposed a model to study the temperatures profiles which are produced in biological tissue when it is heated by RF introducing the effect of the blood perfusion. The electrical-thermal coupled problem has been analytically solved by using the HHTE. COMSOL Multiphysics can be used to solve this kind of problems numerically. We compare the results obtained by COMSOL with those obtained by the analytical solution of the problem.
The link between enthesitis and arthritis in psoriatic arthritis: a switch to a vascular phenotype at insertions may play a role in arthritis development

Objective Subclinical enthesopathy is recognised in both psoriasis and psoriatic arthritis (PsA). This study used ultrasonography with power Doppler (PD) to test the hypothesis that subclinical enthesopathy in PsA was associated with an ‘inflammatory’ or vascular phenotype compared to that seen in psoriasis. Methods 100 patients with a mean age of 46.3 years (SD 15) (42 with psoriasis and 58 with PsA) and 23 matched healthy controls (HC) from two centres were included. 1230 lower limb entheses were scanned by ultrasonographers blinded to clinical details. Both inflammatory and chronic features of enthesopathy were scored. Results Psoriasis patients (with or without arthritis) were more likely to express a vascular phenotype, with higher inflammation-related enthesopathy scores than HC (for inflammation p<0.0001, for chronicity p=0.02, for total ultrasound scores p<0.0001). The PsA patients had higher ultrasound enthesopathy scores than psoriasis patients (inflammation p=0.04, chronicity p=0.02) and HC (inflammation p<0.0001, chronicity p=0.003). When symptomatic entheses were excluded, PsA patients still had higher PD scores than psoriasis patients (p=0.003). Doppler positivity in at least one entheseal site was observed more frequently in PsA (21/58, 36.2%) versus psoriasis (4/42, 9.5%; p=0.002). Conclusions This study shows that the ultrasound appearances of subclinical enthesitis in psoriasis differ from the subclinical enthesitis in PsA, with PsA patients having more PD. This is suggestive of a more inflammatory or vascular process in PsA, and offers potentially novel insights into the progression from skin to joint disease in psoriasis.