Miracle-dz/UniFAR-FTUs huggingface dataset:
Self‐protection and survival of arbovirus‐infected mosquito cells

Arboviruses are serious pathogens for men but cause little damage to their arthropod vectors. We have studied how a mosquito cell line derived from one of the relevant vectors for arboviruses responds to Bunyamwera virus, a well‐characterized arbovirus. Confocal, live cell microscopy and electron microscopy showed that Bunyamwera virus induces deep changes in mosquito cells. Early in infection these cells develop long projections and create new intercellular connections where cell organelles and viral proteins are detected. Live cell microscopy shows that these connections are developed before viral protein can be detected by immunofluorescence. Interestingly, their proliferation is accompanied by a progressive trapping of the nucleocapsid and RNA polymerase viral proteins into large cytoplasmic aggregates. A significant drop in the release of infectious virions then follows. Before that, numerous viruses assemble in peripheral Golgi stacks and they apparently exit the cells immediately since they do not accumulate intracellularly. This mechanism of assembly seems to cause little damage to the integrity of cell endomembranes. The characterization of the antiviral mechanisms operating in mosquito cells can be of great help in the fight against pathogenic arboviruses.
Intravitreal pharmacokinetics and retinal concentrations of ganciclovir and foscarnet after intravitreal administration in rabbits.

PURPOSE To perform a detailed pharmacokinetic study and to evaluate the drug levels reached in the retina after the intravitreal administration of ganciclovir and foscarnet to rabbits. METHODS Retinal and vitreal levels of both drugs were measured by high-performance liquid chromatography at 1, 6, 12, 24, 36, 48, 60, and 72 hours after a single intravitreal injection of 196 microg and 800 microg of ganciclovir and 960 microg of foscarnet to three groups of 24 pigmented rabbits. A noncompartmental pharmacokinetic analysis was used. RESULTS Both drugs incorporated rapidly into the retina, but no equilibrium was observed between the drug levels in the vitreous humor and retina. Mean ganciclovir levels in vitreous and retina were 179.6 microg/g and 131.3 microg/g (dose of 196 microg), 755.7 microg/g and 381.6 microg/g (dose of 800 microg) at 1 hour after administration, decreasing to 0.1 microg/g, 0.6 microg/g, 0.8 microg/g, and 0.7 microg/g, respectively, by 72 hours. Mean foscarnet levels in vitreous and retina were 944 microg/g and 217.1 microg/g at 1 hour after administration, decreasing to 74 microg/g and 17.1 microg/g, respectively, by 72 hours. Whereas both doses of ganciclovir yielded retinal levels above the mean inhibitory concentration (IC50) of most human cytomegalovirus (CMV) isolates for more than 60 hours, foscarnet retinal levels were lower than the CMV IC50 before 36 hours had elapsed after administration. CONCLUSIONS The results suggest that the intravitreal administration of ganciclovir has a better pharmacokinetic profile than foscarnet for the treatment of retinitis caused by CMV and other herpes viruses and support the administration of intravitreal ganciclovir twice a week as a treatment for CMV retinitis.
Cost benefit analyses of alcohol policy - a primer. Final report prepared for the project Standardizing Measurement of Alcohol Related Troubles - SMART.

This paper arises from the project Standardizing Measurement of Alcohol Related Troubles (SMART). 1. Introduction 2. The costs of alcohol policies 2.1. Implementation costs 2.2. Costs of alcohol policy to industry 2.3. Valuing the pleasure of alcohol 3. The benefits of alcohol policy 3.1. Reducing health and welfare costs 3.2. Reducing labour and productivity costs 3.3. Value healthy life 4. Conclusions and summary 5. References
New Progress on the Role of Glia in Iron Metabolism and Iron-Induced Degeneration of Dopamine Neurons in Parkinson’s Disease

It is now increasingly appreciated that glial cells play a critical role in the regulation of iron homeostasis. Impairment of these properties might lead to dysfunction of iron metabolism and neurodegeneration of neurons. We have previously shown that dysfunction of glia could cause iron deposit and enhance iron-induced degeneration of dopamine (DA) neurons in Parkinson’s disease (PD). There also has been a substantial growth of knowledge regarding the iron metabolism of glia and their effects on iron accumulation and degeneration of DA neurons in PD in recent years. Here, we attempt to describe the role of iron metabolism of glia and the effect of glia on iron accumulation and degeneration of DA neurons in the substantia nigra of PD. This could provide evidence to reveal the mechanisms underlying nigral iron accumulation of DA neurons in PD and provide the basis for discovering new potential therapeutic targets for PD.
Subchronic toxicological evaluation of brea gum (Parkinsonia preacox) as a food additive in BALB/c mice

Brea gum is a phloematic exudate from Parkinsonia praecox, an autoctonous tree that grows in the arid areas of Argentina. In this work, we propose its potential as a food additive. However, as no toxicological safety evaluation of brea gum has yet been reported, this preliminary study was conducted to evaluate its long-term toxicity over a 120-day period in BALB/c mice fed with brea gum at various levels in the diet. The results showed that animals on diets containing up to 5% brea gum were healthy, exhibiting growth curves similar to controls for both males (P = 0.9138) and females (P = 0.9459), thereby indicating that feed intake and utilization was not affected. A histopathological examination and weight recording of liver, kidneys, and intestine did not reveal any microscopic abnormalities or adverse toxicological effect (weights respect to control: P > 0.1). Moreover, hematological parameters and enzyme activities were within the normal values previously reported for mice. Our findings suggest that feeding brea gum at levels up to 5% to BALB/c mice do not exert any toxicological effects, supporting its potential use as a food additive for human consumption.
Static Clathrin Assemblies at the Peripheral Vacuole—Plasma Membrane Interface of the Parasitic Protozoan Giardia lamblia

Giardia lamblia is a parasitic protozoan that infects a wide range of vertebrate hosts including humans. Trophozoites are non-invasive but associate tightly with the enterocyte surface of the small intestine. This narrow ecological specialization entailed extensive morphological and functional adaptations during host-parasite co-evolution, including a distinctly polarized array of endocytic organelles termed peripheral vacuoles (PVs), which are confined to the dorsal cortical region exposed to the gut lumen and are in close proximity to the plasma membrane (PM). Here, we investigated the molecular consequences of these adaptations on the Giardia endocytic machinery and membrane coat complexes. Despite the absence of canonical clathrin coated vesicles in electron microscopy, Giardia possesses conserved PV-associated clathrin heavy chain (GlCHC), dynamin-related protein (GlDRP), and assembly polypeptide complex 2 (AP2) subunits, suggesting a novel function for GlCHC and its adaptors. We found that, in contrast to GFP-tagged AP2 subunits and DRP, CHC::GFP reporters have no detectable turnover in living cells, indicating fundamental differences in recruitment to the membrane and disassembly compared to previously characterized clathrin coats. Histochemical localization in electron tomography showed that these long-lived GlCHC assemblies localized at distinctive approximations between the plasma and PV membrane. A detailed protein interactome of GlCHC revealed all of the conserved factors in addition to novel or highly diverged proteins, including a putative clathrin light chain and lipid-binding proteins. Taken together, our data provide strong evidence for giardial CHC as a component of highly stable assemblies at PV-PM junctions that likely have a central role in organizing continuities between the PM and PV membranes for controlled sampling of the fluid environment. This suggests a novel function for CHC in Giardia and the extent of molecular remodeling of endocytosis in this species.
Dopaminergic therapy with carbidopa L-dopa for left neglect after stroke: a case series.

OBJECTIVE To evaluate the efficacy of carbidopa L-dopa (Sinemet) in reducing left spatial neglect after stroke. DESIGN Case series. SETTING Inpatient neurorehabilitation unit in a regional rehabilitation center. PARTICIPANTS A convenience sample of 4 women with right brain strokes and left neglect. INTERVENTION A trial of carbidopa L-dopa to treat left neglect, if indicated by selected subtests of the Behavioral Inattention Test (BIT). MAIN OUTCOME MEASURES Baseline and posttreatment evaluation with the modified BIT and the FIM instrument. RESULTS Three of 4 subjects had significant improvements in their modified BIT scores (8%, 12%, 27%, respectively) and their functional status on the FIM. CONCLUSION With further study, carbidopa L-dopa may be shown to reduce unilateral spatial neglect and thereby improve rehabilitation outcomes.
SUBCORTICAL VASCULAR DISEASE DETECTED WITH COMPUTED TOMOGRAPHY AND 1‐YEAR PHYSICAL DECLINE IN HIGH‐FUNCTIONING OLDER ADULTS

To the Editor: Lopez et al. report that ‘‘cholinesterase inhibitor (CEI) use had a clinically meaningful effect on the natural history of Alzheimer Disease (AD),’’ slowing disease progression and lowering risk of nursing home admission after 2 years. The design of the study is worrisome. Of 1,139 patients who enrolled in the AD Research Center over 7 years, 270 were selected; 135 began taking CEIs ‘‘immediately after enrollment, and continued to take them throughout the following 12 months,’’ and 135 never took the drug. How these individuals were selected is not otherwise described. They were matched on a few characteristics, such as age, Mini-Mental State Examination score, and education. This study resembles a study by Dr. Geldmacher et al. that showed that patients who took a CEI faithfully (80% of pills or more) had a significant delay in nursing home placement (NHP). Both of these nonrandomized studies failed to report important baseline characteristics of the groups being studied. In the Geldmacher article, for example, nonadherent patients were far less likely to have a spouse caregiver than faithful users, yet the authors, who claimed it was the donepezil that ‘‘resulted in significant delays in NHP,’’ omitted this fact. Both papers are easily distinguished from AD 2000, a properly randomized, controlled trial with the largest number of placebo-controlled patient-years of any cholinesterase study. In AD 2000, ‘‘no significant benefits were seen with donepezil compared with placebo in institutionalization or progression of disability . [or] in behavioral and psychological symptoms, carer psychopathology, formal care costs, unpaid caregiver time, adverse events, or death or between 5 mg and 10 mg of donepezil. ’’ In the United Kingdom, the National Institute for Clinical Excellence Appraisal Committee has recently issued the following preliminary recommendation: ‘‘Donepezil, rivastigmine and galantamine are not recommended for use in the treatment of mild to moderate Alzheimer’s disease (AD). ’’ Would Dr. Lopez modify the discussion of his paper, where he emphasizes the important benefits of donepezil, in view of the results from AD 2000, a larger, better-designed trial that failed to show any meaningful difference at all?
Electron microscopy studies of the effects of garlic extract against Trichophyton rubrum

Trichophyton rubrum is one of dermatophytes that penetrates keratinized tissues such as skin, hair and nail of human and animals. Recently, antifungal drugs such as imodazole and triazole was found to cause side effects, toxicity to patients and also not very efficient due to resistance to these drugs. As an alternative, some plants extract had been used to treat dermatophytes. This studies was done using Garlic extract (Allium sativum) to evaluate its effects on the growth of hypha of Trichophyton using Electron miscroscopy. Garlic had been known to posses antimicrobial, antiinflammatory, antithrombotic and antitumor activities. This studies found that garlic extract as low as 4 mg/mL inhibit the growth of hypha. Scanning electron microscopy studies revealed that hypha treated with garlic extract showed shrinkage, flat and cell wall demolition, similar to hypha treated with allicin (positive control) having rough surface, shrinkage and distortion. The tip of hypa became large after treatment with garlic extract. Transmission electron microscopy studies also found that hypha treated with allicin display cell wall thickening, local thickening, destruction of cytoplasmic content, mean while hypha treated with garlic extract exhibited cell wall thickening, disordered hyphal tip and desolution of cytoplasmic compartments and similar with hypha treated with allicin. These results showed that garlic extract and pure allicin could be use as an alternative to treat dermatophytes.
Multicentre Evaluation of an Enzyme-Immunoassay for Cortisol Determination

The present paper describes a multicentre evaluation of a one step enzyme-immunoassay for the determination of cortisol in serum or plasma. Data from the investigation were analysed in terms of imprecision, detection limit, and correlation with other test methods. Within-run and between-run imprecisions (coefficient of variation) of Enzymun-Test Cortisol were less than 8% and 12%, respectively. The detection limit was 30 nmol/l (11 micrograms/l). With the exception of prednisolone, only low interference was found with other endogenous steroids. A good correlation between Enzymun-Test Cortisol and HPLC, LIA, FPIA and RIA was registered, although the latter two methods showed a scattering of regression lines from the different evaluators. The results show that Enzymun-Test Cortisol can be recommended as an alternative for the measurement of cortisol. As the method is calibrated against isotope dilution-mass spectrometry, results obtained with Enzymun-Test Cortisol are in agreement with the reference method.
EFFECTS OF EXERCISE AND ADMINISTRATION OF ASPARTIC ACID ON BLOOD AMMONIA IN THE RAT.

The rise of blood ammonia following a short period of exercise can be partially prevented by the prior oral administration of either aspartic acid or the mixed potassium and magnesium salts of aspartic acid. These compounds will also prolong the time required for the rat to reach physical exhaustion due to forced swimming. As an extension of these observations, it has been found that daily feeding of ammonium carbonate over a 4-week period will also prevent the rise of blood ammonia due to exercise and will prolong exhaustion time of rats that are forced to swim. These same changes have been observed in rats that were exercised daily by short periods of forced swimming and, after 3 weeks of this type of physical training, were tested for blood ammonia response to exercise and time required for exhaustion. It is concluded that both daily ammonium carbonate feeding and training result in adaptation of enzymes in the ornithine cycle so as to increase the rate of blood ammonia removal.
Use of in vivo Expression Technology for the Identification of Putative Host Adaptation Factors of the Lyme Disease Spirochete

The causative agent of Lyme disease, Borrelia burgdorferi, is an obligate parasite that requires either a tick vector or a mammalian host for survival. Identification of the bacterial genes that are specifically expressed during infection of the mammalian host could provide targets for novel therapeutics and vaccines. In vivo expression technology (IVET) is a reporter-based promoter trap system that utilizes selectable markers to identify promoters of bacterial host-specific genes. Using previously characterized genes for in vivo and in vitro selection, this study utilized an IVET system that allows for selection of B. burgdorferi sequences that act as active promoters only during murine infection. This promoter trap system was able to successfully distinguish active promoter sequences both in vivo and in vitro from control sequences and a library of cloned B. burgdorferi genomic fragments. However, a bottleneck effect during the experimental mouse infection limited the utility for genome-wide promoter screening. Overall, IVET was demonstrated as a tool for the identification of in vivo-induced promoter elements of B. burgdorferi, and the observed infection bottleneck apparent using a polyclonal infection pool provides insight into the dynamics of experimental infection with B. burgdorferi.
Functions of Cell Surface-Anchored Antigen I/II Family and Hsa Polypeptides in Interactions of Streptococcus gordonii with Host Receptors

ABSTRACT Streptococcus gordonii colonizes multiple sites within the human oral cavity. This colonization depends upon the initial interactions of streptococcal adhesins with host receptors. The adhesins that bind salivary agglutinin glycoprotein (gp340) and human cell surface receptors include the antigen I/II (AgI/II) family polypeptides SspA and SspB and a sialic acid-binding surface protein designated Hsa or GspB. In this study we determined the relative functions of the AgI/II polypeptides and Hsa in interactions of S. gordonii DL1 (Challis) with host receptors. For an isogenic mutant with the sspA and sspB genes deleted the levels of adhesion to surface-immobilized gp340 were reduced 40%, while deletion of the hsa gene alone resulted in >80% inhibition of bacterial cell adhesion to gp340. Adhesion of S. gordonii DL1 cells to gp340 was sialidase sensitive, verifying that Hsa has a major role in mediating sialic acid-specific adhesion to gp340. Conversely, aggregation of S. gordonii cells by fluid-phase gp340 was not affected by deletion of hsa but was eliminated by deletion of the sspA and sspB genes. Deletion of the AgI/II polypeptide genes had no measurable effect on hsa mRNA levels or Hsa surface protein expression, and deletion of hsa did not affect AgI/II polypeptide expression. Further analysis of mutant phenotypes showed that the Hsa and AgI/II proteins mediated adhesion of S. gordonii DL1 to human HEp-2 epithelial cells. Hsa was also a principal streptococcal cell surface component promoting adhesion of human platelets to immobilized streptococci, but Hsa and AgI/II polypeptides acted in concert in mediating streptococcal cell-platelet aggregation. The results suggest that Hsa directs primary adhesion events for S. gordonii DL1 (Challis) with immobilized gp340, epithelial cells, and platelets. AgI/II polypeptides direct gp340-mediated aggregation, facilitate multimodal interactions necessary for platelet aggregation, and modulate S. gordonii-host engagements into biologically productive phenomena.
Effects of Acute and Repeated Administration of Oxycodone and Naloxone-Precipitated Withdrawal on Intracranial Self-Stimulation in Rats

Incidence of prescription opioid abuse and overdose, often led by oxycodone, continues to increase, producing twice as many overdose deaths as heroin. Surprisingly, preclinical reports relevant to oxycodone’s abuse-related effects are relatively sparse considering its history and patient usage. The goal of this study was to characterize dose- and time-dependent effects of acute and repeated oxycodone administration in a frequency-rate intracranial self-stimulation (ICSS) procedure, an assay often predictive of drug-related reinforcing effects, in male Sprague-Dawley rats. We hypothesized that oxycodone would produce a biphasic profile of rate-increasing and rate-decreasing effects maintained by ICSS similar to μ-opioid receptor agonists. Oxycodone (0.03, 0.3, 1, and 3 mg/kg, s.c.) produced dose- and time-dependent alterations on ICSS, with the predicted biphasic profile of rate-increasing effects at lower stimulation frequencies followed by rate-decreasing effects at higher frequencies. Peak effects were observed between 30 and 60 minutes, which were reversed by naloxone pretreatment (30 minutes). Tolerance to rate-decreasing effects was observed over a 5-day period when rats were treated with 1 mg/kg oxycodone twice a day. Subsequently, the dosing regimen was increased to 3 mg/kg twice a day over 10 days, although further marked tolerance did not develop. When then challenged with 10 mg/kg naloxone, a significant suppression below baseline levels of ICSS-maintained responding occurred indicative of dependence that recovered to baseline within 5 hours. The results of this study provide the first report of acute and chronic effects of oxycodone on responding maintained by ICSS presentation and the use of ICSS-maintained responding to characterize its tolerance and dependence effects.
Haemoglobin Level and Intestinal Parasites in Pregnant Women in Eziama, Imo State of Nigeria

This prospective study was carried out between January to June, 2008 at Eziama in Isiala Mbano Local Government Area of Imo State to determine the haemoglobin level and intestinal parasites in pregnant women attending antenatal clinic. In total, 281 stool and blood samples were collected from the women. The stool samples were examined using direct wet preparation and formol ether concentration technique while the haemoglobin levels of the women were estimated using cyanmethaeoglobin technique. The overall prevalence of E. histolytical, A. Lumbricoides, Hookworm Spp, T. trichuria and G. lamblia was 10(3.6), 31(11.0), 29(10.3), 10(3.6) and 3(1.1) respectively. The proportion of pregnant women with helminithic infestation was 83 (29.6%) while their mean Heamoglobin concentration was 9.7+1.8g/dl while the pregnant women without helminthic infestation had Heamoglobin concentration of 10.4 + 1.3g/dl. The poor socioeconomic status of the women coupled with poor environmental sanitation and lack of clean portable water supply may have contributed to the high prevalence.
Development of the AGREE II, part 2: assessment of validity of items and tools to support application

Background: We established a program of research to improve the development, reporting and evaluation of practice guidelines. We assessed the construct validity of the items and user’s manual in the β version of the AGREE II. Methods: We designed guideline excerpts reflecting high-and low-quality guideline content for 21 of the 23 items in the tool. We designed two study packages so that one low-quality and one high-quality version of each item were randomly assigned to each package. We randomly assigned 30 participants to one of the two packages. Participants reviewed and rated the guideline content according to the instructions of the user’s manual and completed a survey assessing the manual. Results: In all cases, content designed to be of high quality was rated higher than low-quality content; in 18 of 21 cases, the differences were significant (p < 0.05). The manual was rated by participants as appropriate, easy to use, and helpful in differentiating guidelines of varying quality, with all scores above the mid-point of the seven-point scale. Considerable feedback was offered on how the items and manual of the β-AGREE II could be improved. Interpretation: The validity of the items was established and the user’s manual was rated as highly useful by users. We used these results and those of our study presented in part 1 to modify the items and user’s manual. We recommend AGREE II (available at www.agreetrust.org) as the revised standard for guideline development, reporting and evaluation.