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Trypanosoma cruzi has three biochemically and morphologically distinct developmental stages that are programmed to rapidly respond to environmental changes the parasite faces during its life cycle. Unlike other eukaryotes, Trypanosomatid genomes contain protein coding genes that are transcribed into polycistronic pre-mRNAs and have their expression controlled by post-transcriptional mechanisms. Transcriptome analyses comparing three stages of the T. cruzi life cycle revealed changes in gene expression that reflect the parasite adaptation to distinct environments. Several genes encoding RNA binding proteins (RBPs), known to act as key post-transcriptional regulatory factors, were also differentially expressed. We characterized one T. cruzi RBP, named TcZH3H12, which contains a zinc finger domain and is up-regulated in epimastigotes compared to trypomastigotes and amastigotes. TcZC3H12 knockout (KO) epimastigotes showed decreased growth rates and increased capacity to differentiate into metacyclic trypomastigotes. Transcriptome analyses comparing wild type and TcZC3H12 KOs revealed a TcZC3H12-dependent expression of epimastigote-specific genes such as genes encoding amino acid transporters and proteins associated with differentiation (PADs). RNA immunoprecipitation assays showed that transcripts from the PAD family interact with TcZC3H12. Taken together, these findings suggest that TcZC3H12 positively regulates the expression of genes involved in epimastigote proliferation and also acts as a negative regulator of metacyclogenesis.
Intermittent food restriction (IFR) is used mainly for weight loss; however, its effects on adipose tissue are not known when alternating with an obesogenic diet. To demonstrate its effects on morphological dynamics of fat deposits, female Wistar rats were distributed into groups: standard control (ST-C), with commercial diet; DIO control (DIO-C), with a diet that induces obesity (DIO) during the first and last 15 d, replaced by a standard diet for thirty intermediate days; standard restricted (ST-R), with standard diet during the first and last 15 d, with six cycles of IFR at 50 % of ST-C; and DIO restricted (DIO-R), in DIO during the first and last 15 d, with six cycles of IFR at 50 % of DIO-C. At 105 d of life, white adipose tissue (WAT) and brown adipose tissue (BAT) deposits were collected, weighed and histology performed. The DIO-R group showed higher total food intake (DIO-R 10 768·0 (SEM 357·52) kJ/g v. DIO-C 8868·6 (SEM 249·25) kJ/g, P < 0·0001), energy efficiency during RAI (DIO-R 2·26 (SEM 0·05) g/kJ v. DIO-C 0·70 (SEM 0·03) g/kJ, P < 0·0001) and WAT (DIO-R 5·65 (SEM 0·30) g/100 g v. DIO-C 4·56 (SEM 0·30) g/100 g) than their respective control. Furthermore, IFR groups presented hypertrophy of WAT and BAT, as well as fibrosis in BAT. Thus, IFR can establish prospective resistance to weight loss by favouring changes in adipose tissue morphology, increased energy intake and efficiency. Finally, the DIO diet before and after IFR aggravates the damages caused by the restriction.
The co-infection between visceral leishmaniasis (VL) and human immunodeficiency virus (HIV) has increased in several countries in the world. The current serological tests are not suitable since they present low sensitivity to detect the most of VL/HIV cases, and a more precise diagnosis should be performed. In this context, in the present study, an immunoproteomics approach was performed using Leishmania infantum antigenic extracts and VL, HIV and VL/HIV patients sera, besides healthy subjects samples; aiming to identify antigenic markers for these clinical conditions. Results showed that 43 spots were recognized by antibodies in VL and VL/HIV sera, and 26 proteins were identified by mass spectrometry. Between them, β-tubulin was expressed, purified and tested in ELISA experiments as a proof of concept for validation of our immunoproteomics findings and results showed high sensitivity and specificity values to detect VL and VL/HIV patients. In conclusion, the identified proteins in the present work could be considered as candidates for future studies aiming to improvement of the diagnosis of VL and VL/HIV co-infection.
To describe a case of conversion disorder in a pre-school girl, affecting the sensory functions.
Materials and methods
The authors revise the specific characteristics of this disorder in pre-school children and highlight its differences from adult's common pictures. With that purpose they describe a case of a 5 year's old girl who attended the ER with sudden blindness complaints. She was admitted for observation and investigation. All possible organic causes where excluded. She was then referred to Child and Adolescent Psychiatry emergency consultation. She was diagnosed with Conversion Disorder triggered by a familiar stressful situation and was medicated with diazepam. Her vision recovered suddenly within a few hours.
Results and conclusions
Although common among adolescents and older children, conversion disorder is a rare entity among pre-school children. The affected function in this age group is usually the motor function, but in rare cases it can affect the sensory functions causing deafness or blindness. This seems to be associated with stressful events but also has a strong familiar component, where imitation of adult's behaviors and familiar gains take major roles.
Self-injurious behaviors(SIB) may be described as a behavior compatible with obsessive compulsive disorder(OCD), as well as part of Impulse-Control Disorders(ICD).
Compare obsessive compulsive symptoms(OCS) and impulsivity between two groups: OCD and SIB patients.
Five patients in outpatient treatment for SIB were compared to five OCD patients. The following instruments were applied:Y-BOCS, D-YBOCS, USP-SPS, FASM, SCID, BIS-11.
The SIBs were repetitive and occurred from 2 to 100 episodes during the last year. The behavior relief intolerable affects and the consequent pain were mild. The patients spent some time planning the acts, especially when it was not possible to do it immediately. The most common behavior found were: skin cutting(80%), self-hitting(60%), self-biting(60%), and the patients presented more than one type of SIBs. The motivation included: relieving feelings of "numbness" (80%); punishing themselves(80%); feeling something (even pain)(60%); feeling relaxed(60%); and stopping bad feelings(60%). SIB patients also presented OCS(100%) with more prevalence of sensory phenomena preceding repetitive behaviors(100%) than among OCD patients(60%), although they were not statistically significant. Despite the fact that there were no difference between the two groups in impulsivity according to BIS-11(66,20±18,10 SD for SIB and 68,40±11,10 SD for OCD, p=0,82), SIB patients had more diagnosis of others ICD(100%).
This study raises the question: Would SIB be an OCS, that increases it severity, or a nosologic entity with its own characteristics, where OCD comobidity is frequent? The sample size is not large enough to answer these questions, although it seems that some symptoms are shared with both disorders.
Self-mutilation behavior (SMB) is defined as all behaviors involving deliberate infliction of direct physical harm to one's own body without any intent to die. This case report describes the successful treatment of severe SMB in a 23-year-old woman, with multiple comorbidities. The patient was admitted to the multiple impulse-control disorder outpatient unit for treatment of SMB. This patient was submitted to the SCID-I/P, SCID-II/P, Y-BOCS, DY-BOCS, and Functional Assessment of Self-Mutilation (FASM) for diagnosis of SMB as well as comorbidities. The most frequently SMB presented was skin cutting which was associated with relief of intolerable affects. Others comorbidities presented by her were Obsessive-Compulsive Disorder(OCD), Social phobia, Bulimia, and Depression Disorder(DD) with high levels of anxiety. The patient was submitted to an interdisciplinary treatment. Treatment included cognitive-behavior therapy(CBT), nutritional orientation, and psychopharmacology which begun with venlafaxine (150mg/d) followed by fluoxetine(80mg/day), and carbamazepine(400mg/day), without success. After ten months of treatment the patient was stabilized in terms of DD, bulimic behaviors, and anxiety. OCD symptoms were mild, and SMB became rare. At this time the patient was taking ziprazidone(160mg/day), sertraline(200mg/day), and topiramate(100mg/day). These medications were maintained as well as CBT, with emphasis in social skill, and problem-solving techniques. After three more months of treatment she had stopped cutting herself and the OCD symptoms disappeared.
the collection of treatment modalities implemented by multidisciplinary team may serve as a guide to treat severe SMB. In addition, the association of drugs with different site of action, but all for impulse control, may contribute for the efficacy observed here.
Non-Suicidal Self-Injury (NSSI) is considered a dysfunctional way of dealing with problem situations.
This study aimed to investigate the problem solving capacity in adults with NSSI compared to controls.
Thirty-three patients who sought treatment for NSSI (NSSI group) were compared with 33 individuals without psychiatric disorder (control group). We also investigated Axis I disorders, executive functions and problem solving capacity.
In both groups, the majority were women (77.25%) with a mean age of 30 years, and the beginning of NSSI behavior of 16 years. The most common NSSI behaviour was skin cutting, and the most common reason given for engaging in that behaviour was “to stop negative feelings”. The most common psychiatric comorbidities were major depressive disorder (60.6%). Compared to controls, the group with NSSI showed lower results in relation to problem solving capacity (P = 0.000) and mental flexibility (P = 0.007). Deficits in problem solving capacity may be a reflection of low mental flexibility of adults with NSSI. This may be a risk factor for the beginning of NSSI and the persistence of it in adulthood.
Early identification and treatment focused on problem solving capacity during the adolescence may prevent the chronicity of NSSI.
Disclosure of interest
The authors have not supplied their declaration of competing interest.
Barramundi (Lates calcarifer) are a highly valued aquaculture species, and, as obligate carnivores, they have a demonstrated preference for dietary protein over lipid or starch to fuel energetic growth demands. In order to investigate how carnivorous fish regulate nutritional cues, we examined the metabolic effects of feeding two isoenergetic diets that contained different proportions of digestible protein or starch energy. Fish fed a high proportion of dietary starch energy had a higher proportion of liver SFA, but showed no change in plasma glucose levels, and few changes in the expression of genes regulating key hepatic metabolic pathways. Decreased activation of the mammalian target of rapamycin growth signalling cascade was consistent with decreased growth performance values. The fractional synthetic rate (lipogenesis), measured by TAG 2H-enrichment using 2H NMR, was significantly higher in barramundi fed with the starch diet compared with the protein diet (0·6 (se 0·1) v. 0·4 (se 0·1) % per d, respectively). Hepatic TAG-bound glycerol synthetic rates were much higher than other closely related fish such as sea bass, but were not significantly different (starch, 2·8 (se 0·3) v. protein, 3·4 (se 0·3) % per d), highlighting the role of glycerol as a metabolic intermediary and high TAG-FA cycling in barramundi. Overall, dietary starch significantly increased hepatic TAG through increased lipogenesis. Compared with other fish, barramundi possess a unique mechanism to metabolise dietary carbohydrates and this knowledge may define ways to improve performance of advanced formulated feeds.
Patient assessment is a fundamental feature of community paramedicine, but the absence of a recognized standard for assessment practices contributes to uncertainty about what drives care planning and treatment decisions. Our objective was to summarize the content of assessment instruments and describe the state of current practice in community paramedicine home visit programs.
We performed an environmental scan of all community paramedicine programs in Ontario, Canada, and used content analysis to describe current assessment practices in home visit programs. The International Classification on Functioning, Disability, and Health (ICF) was used to categorize and compare assessments. Each item within each assessment form was classified according to the ICF taxonomy.
A total of 43 of 52 paramedic services in Ontario, Canada, participated in the environmental scan with 24 being eligible for further investigation through content analysis of intake assessment forms. Among the 24 services, 16 met inclusion criteria for content analysis. Assessment forms contained between 13 and 252 assessment items (median 116.5, IQR 134.5). Most assessments included some content from each of the domains outlined in the ICF. At the subdomain level, only assessment of impairments of the functions of the cardiovascular, hematological, immunological, and respiratory systems appeared in all assessments.
Although community paramedicine home visit programs may differ in design and aim, all complete multi-domain assessments as part of patient intake. If community paramedicine home visit programs share similar characteristics but assess patients differently, it is difficult to expect that the resulting referrals, care planning, treatments, or interventions will be similar.
There is no suitable vaccine against human visceral leishmaniasis (VL) and available drugs are toxic and/or present high cost. In this context, diagnostic tools should be improved for clinical management and epidemiological evaluation of disease. However, the variable sensitivity and/or specificity of the used antigens are limitations, showing the necessity to identify new molecules to be tested in a more sensitive and specific serology. In the present study, an immunoproteomics approach was performed in Leishmania infantum promastigotes and amastigotes employing sera samples from VL patients. Aiming to avoid undesired cross-reactivity in the serological assays, sera from Chagas disease patients and healthy subjects living in the endemic region of disease were also used in immunoblottings. The most reactive spots for VL samples were selected, and 29 and 21 proteins were identified in the promastigote and amastigote extracts, respectively. Two of them, endonuclease III and GTP-binding protein, were cloned, expressed, purified and tested in ELISA experiments against a large serological panel, and results showed high sensitivity and specificity values for the diagnosis of disease. In conclusion, the identified proteins could be considered in future studies as candidate antigens for the serodiagnosis of human VL.
Introduction: Patient assessment is a fundamental feature of non-emergency community paramedicine (CP) home visit programs. In the absence of a recognized standard for CP assessment, current assessment practices in CP programs are unknown. Without knowing what community paramedics are assessing, it is difficult to ascertain what should be included in patient care plans, whether interventions are beneficial, or whether paramedics are meeting program objectives. Our objective was to summarize the content of assessment instruments used in CP programs in order to describe the state of current practice. Methods: We performed an environmental scan of all CP programs in Ontario, Canada, and employed content analysis to describe current assessment practices in CP home visit programs. The International Classification on Functioning, Disability, and Health (ICF) was used to categorize and compare assessments. Each item within each assessment form was classified according to the ICF taxonomy. Findings were compared at the domain and sub-domain of the ICF. Results: Of 54 paramedic services in Ontario, 43 responded to our request for information. Of 24 services with CP home visit programs, 18 provided their intake assessment forms for content analysis. Assessment forms contained between 13 and 252 assessment items (median 116.5, IQR 134.5). Overall, most assessments included some content from each of the domains outlined in the ICF, including: Impairments of Body Functions, Impairments of Body Structures, Activity Limitation and Participation, and Environmental Factors. At the sub-domain level, only assessment of Impairments of the Functions of the Cardiovascular, Haematological, Immunological and Respiratory systems appeared in all assessments. Few CP home visit program assessments covered most ICF sub-domain categories and many items classified to specific categories were included in only a few assessments. Conclusion: CP home visit programs complete multi-domain assessments as part of patient intake. The content of CP assessments varied across Ontario, which suggests that care planning and resources may not be consistent. Current work on practice guidelines and paramedic training can build from descriptions of assessment practices to improve quality of care and patient safety. By identifying what community paramedics assess, evaluation of the quality of CP home visit programs and their ability to meet program objectives can be improved and benchmarks in patient care can be established.
The adsorption of intact liposomes on surfaces is of great importance for the development of sensors and drug delivery systems and, also, strongly dependent on the surface roughness where the liposomes are adsorbed. In this paper, we analyzed, by using atomic force microscopy in liquid, the evolution of the morphology of gold surfaces and of poly(allylamine hydrochloride) (PAH) surfaces with different roughness during the adsorption of liposomes prepared with the synthetic phospholipid 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)]. Our results reveal the following. On smooth surfaces of Au only and Au with PAH, the liposomes open and deploy on the substrate, creating a supported-lipid bilayer, with the opening process being faster on the Au/PAH surface. On rough substrates of Au coated with polyelectrolyte multilayers, the liposomes were adsorbed intact on the surface. This was corroborated by power spectral density analysis that demonstrates the presence of superstructures with an average lateral size of 43 and 87 nm, in accordance with two and four times the mean liposome hydrodynamic diameter of about 21 nm. In addition, this work presents an adequate and effective methodology for analysis of adsorption phenomena of liposomes on rough surfaces.
In this paper we provide an introduction to statistical inference for the classical linear birth‒death process, focusing on computational aspects of the problem in the setting of discretely observed processes. The basic probabilistic properties are given in Section 2, focusing on computation of the transition functions. This is followed by a brief discussion of simulation methods in Section 3, and of frequentist methods in Section 4. Section 5 is devoted to Bayesian methods, from rejection sampling to Markov chain Monte Carlo and approximate Bayesian computation. In Section 6 we consider the time-inhomogeneous case. The paper ends with a brief discussion in Section 7.
In this study, a Leishmania hypothetical protein, LiHyS, was evaluated regarding its antigenicity, immunogenicity and protective efficacy against visceral leishmaniasis (VL). Regarding antigenicity, immunoblottings and an enzyme-linked immunosorbent assay using human and canine sera showed high sensitivity and specificity values for the recombinant protein (rLiHyS) in the diagnosis of VL. When evaluating the immunogenicity of LiHyS, which is possibly located in the parasite's flagellar pocket, proliferative assays using peripheral blood mononuclear cells from healthy subjects or VL patients showed a high proliferative index in both individuals, when compared to the results obtained using rA2 or unstimulated cultures. Later, rLiHyS/saponin was inoculated in BALB/c mice, which were then challenged with Leishmania infantum promastigotes. The vaccine induced an interferon-γ, interleukin (IL)-12 and granulocyte-macrophage colony-stimulating factor production, which was maintained after infection and which was associated with high nitrite and IgG2a antibody levels, as well as low IL-4 and IL-10 production. Significant reductions in the parasite load in liver, spleen, bone marrow and draining lymph nodes were found in these animals. In this context, the present study shows that the rLiHyS has the capacity to be evaluated as a diagnostic marker or vaccine candidate against VL.
The rheological behavior of composites made with high-density polyethylene (HDPE) and chitosan was studied. Composites were prepared by melt processing in a laboratory internal mixer. Maleic anhydride grafted HDPE (PE-g-MA) was used as compatibilizer to enhance the dispersion of chitosan in the HDPE matrix. Different percentages of chitosan and compatibilizer (up to a maximum of 25 phr) were added into HDPE to prepare composites. Characterization of the composites with parallel plate rheometer and laboratory internal mixer revealed that the presence of chitosan increases the complex viscosity, loss modulus, storage modulus and the torque (i.e., melt viscosity), and the combination chitosan/compatibilizer has a similar, if slighter, effect. At higher filler levels it is clear that the PE-g-MA affected the microstructure of the compounds, possibly increasing matrix–filler interactions and acting as an effective compatibilizer.
Paramedicine is experiencing significant growth in scope of practice, autonomy, and role in the health care system. Despite clinical governance models, the degree to which paramedicine ultimately can be safe and effective will be dependent on the individuals the profession deems suited to practice. This creates an imperative for those responsible for these decisions to ensure that assessments of paramedic competence are indeed accurate, trustworthy, and defensible.
The purpose of this study was to explore and synthesize relevant theoretical foundations and literature informing best practices in performance-based assessment (PBA) of competence, as it might be applied to paramedicine, for design or evaluation of assessment programs.
A narrative review methodology was applied to focus intentionally, but broadly, on purpose relevant, theoretically derived research that could inform assessment protocols in paramedicine. Primary and secondary studies from a number of health professions that contributed to and informed best practices related to the assessment of paramedic clinical competence were included and synthesized.
Multiple conceptual frameworks, psychometric requirements, and emerging lines of research are forwarded. Seventeen practice implications are derived to promote understanding as well as best practices and evaluation criteria for educators, employers, and/or licensing/certifying bodies when considering the assessment of paramedic competence.
The assessment of paramedic competence is a complex process requiring an understanding, appreciation for, and integration of conceptual and psychometric principles. The field of PBA is advancing rapidly with numerous opportunities for research.
TavaresW, BoetS. On the Assessment of Paramedic Competence: A Narrative Review with Practice Implications. Prehosp Disaster Med. 2016;31(1):64–73.
Two mimotopes of Leishmania infantum identified by phage display were evaluated as vaccine candidates in BALB/c mice against Leishmania amazonensis infection. The epitope-based immunogens, namely B10 and C01, presented as phage-fused peptides; were used without association of a Th1 adjuvant, and they were administered isolated or in combination into animals. Both clones showed a specific production of interferon-gamma (IFN-γ), interleukin-12 (IL-12) and granulocyte/macrophage colony-stimulating factor (GM-CSF) after in vitro spleen cells stimulation, and they were able to induce a partial protection against infection. Significant reductions of parasite load in the infected footpads, liver, spleen, bone marrow and paws’ draining lymph nodes were observed in the immunized mice, in comparison with the control groups (saline, saponin, wild-type and non-relevant clones). Protection was associated with an IL-12-dependent production of IFN-γ, mediated mainly by CD8+ T cells, against parasite proteins. Protected mice also presented low levels of IL-4 and IL-10, as well as increased levels of parasite-specific IgG2a antibodies. The association of both clones resulted in an improved protection in relation to their individual use. More importantly, the absence of adjuvant did not diminish the cross-protective efficacy against Leishmania spp. infection. This study describes for the first time two epitope-based immunogens selected by phage display technology against L. infantum infected dogs sera, which induced a partial protection in BALB/c mice infected with L. amazonensis.
We present a multifrequency analysis of the variability in the flat-spectrum radio quasar 3C 279 from 2008 to 2014. Our multiwavelength dataset includes gamma-ray data from Fermi/LAT (Abdo et al. 2009), observations in 1mm from SMA (Gurwell et al. 2007), Near Infrared from OAGH (Carramiñana & Carrasco 2009) and SMARTS (Bonning et al. 2012); optical V band from the Steward Observatory (Smith et al. 2009) and SMARTS; optical spectra from OAGH (Patiño-Álvarez et al. 2013) and the Steward Observatory; and polarization spectra from the Steward Observatory. The light curves are shown in Fig. 1. Six out of seven optical activity periods identified within our dataset show clear counterparts in mm, NIR and gamma-rays, however, the late 2011 - early 2012 optical flare does not have a counterpart in the GeV regime. In this contribution, we discuss the flaring evolution of 3C 279 and speculate about the production of the anomalous activity period.