Analysis of clinical outcomes in older individuals who received pharmaceutical care and posthospital discharge follow-up.

BACKGROUND: Previous evidence underscores the need to assess potential clinical outcomes resulting from pharmaceutical care interventions and to monitor patient's progress to evaluate their clinical evolution, which is crucial for bolstering the relevance of implementing pharmaceutical care in healthcare services. AIMS: To conduct an in-depth analysis of pharmaceutical care practices in a geriatric ward and monitor the clinical outcomes of older people served. METHODS: This interventionist study was conducted in the geriatrics ward of a Brazilian university hospital. The research intervention occurred between January and May 2022, with a follow-up conducted for up to 90 days after patients' hospital discharge. Older patients in the geriatrics ward received pharmaceutical care, including medication reconciliation, medication review, and pharmacotherapeutic follow-up, aimed at identifying and resolving drug-related problems (DRPs). The clinical relevance of DRPs and pharmaceutical recommendations was evaluated. Additionally, analyses were conducted on mortality and rehospitalization outcomes in older patients at 30, 60, and 90 days following initial hospital discharge. RESULTS: Of the patients evaluated, a significant 88.3% exhibited at least one DRP (with an average of 2.6 ± 1.9 DRPs per patient), with the majority classified as need/indication problems (38.9%). The acceptance rate of pharmaceutical recommendations was 80.9%, with the majority categorized as very significant relevance (60.4%). DRPs were predominantly of serious clinical relevance (50.9%). In patients whose clinical indicators could be monitored, 95.5% showed some clinical response (in vital signs, laboratory tests and/or clinical status evolution) potentially related to resolved DRPs. Association analysis revealed that a higher number of medications in use before hospitalization correlated with a greater identification of DRPs during hospitalization (p = 0.03). At hospital discharge, 23.6% of patients were no longer using polypharmacy. In total, 16 patients (26.7%) died during the study period. Among patients who did not die during hospitalization (n = 54), 20 patients (37%) experienced rehospitalizations within 90 days following discharge. CONCLUSION: This study facilitated the consolidation of pharmaceutical care implementation in a geriatric ward. We conducted identification, evaluation, and proposed evidence-based solutions, as well as monitored cases for outcome analysis. It is anticipated that this methodology will inspire future research and the implementation of pharmaceutical care-related services.

Antinociceptive and anti-inflammatory potential, and chemical characterization of the dichloromethane fraction of Solanum lycocarpum (Solanaceae) ripe fruits by LC-DAD-MS.

ETHNOPHARMACOLOGICAL RELEVANCE: Solanum lycocarpum A. St. Hil. (Solanaceae) is a species from the Brazilian Cerrado, exhibiting several medicinal properties, being used by the population in the treatment of ulcers, bronchitis, asthma and hepatitis, which involve inflammatory processes. AIM OF THIS STUDY: This study aimed to chemically characterize the dichloromethane fraction (DCM), as well as verify its antinociceptive, anti-inflammatory and antioxidant potential. MATERIALS AND METHODS: The DCM fraction was obtained by partitioning the ethanol extract. The chemical constituents of the DCM fraction were characterized by LC-DAD-MS. The DPPH and FRAP assays were used to evaluate the antioxidant potential. The carrageenan-induced paw edema model was used to assess the anti-inflammatory effects, and the inflammatory infiltrate was evaluated by qualitative and quantitative histological analyses. The antinociceptive action of the DCM fraction was evaluated by acetic acid-induced abdominal writhing test, formalin-induced nociception and hot-plate test. RESULTS: Steroidal alkaloids solasonine, solasodine and solamargine, as well as the alkaloid peiminine/imperialine and caffeoylquinic acids, were annotated in DCM fraction by LC-DAD-MS. The DCM fraction showed antioxidative action in the in vitro DPPH and FRAP tests, as well as an anti-inflammatory effect for the three evaluated doses of 30, 100 and 300 mg/kg in the fourth and sixth hours after the administration of carrageenan. The histological analyses evidenced considerably reduction in leukocyte migration and the number of polymorphonuclear leukocytes. The study also demonstrated antinociceptive activity for the DCM fraction, which reduced abdominal writhing at three concentrations evaluated, as well as a decrease in paw licking in the formalin-induced nociception test both in the neurogenic phase and the inflammatory phase, with greater effectiveness compared to the anti-inflammatory indomethacin. The DCM fraction also increased the latency time of the animals in the hot plate test 60 min after treatment, although it did not seem to involve the opioidergic system. CONCLUSION: This work evidenced that the dichloromethane fraction of S. lycocarpum fruit possesses antinociceptive and anti-inflammatory potential, which supports its use in folk medicine for management inflammatory conditions.

Isolation of saprophytic filamentous fungi from avian fecal samples and assessment of its predatory activity on coccidian oocysts.

Fungal strains used in the biocontrol of animal gastrointestinal parasites have been mainly isolated from pasture soil, decaying organic matter, and feces from herbivores and carnivores. However, their isolation from birds and assessment of predatory activity against avian GI parasites has been scarce thus far. This research aimed to isolate filamentous fungi from avian fecal samples and evaluate their predatory activity against coccidia. A pool of 58 fecal samples from chickens, laying hens, and peacocks, previously collected between July 2020-April 2021, were used for isolation of filamentous fungi and assessment of their in vitro predatory activity against coccidian oocysts, using Water-Agar medium and coprocultures. The Willis-flotation technique was also performed to obtain concentrated suspensions of oocysts. A total of seven Mucor isolates was obtained, being the only fungal taxa identified, and all presented lytic activity against coccidia. Isolates FR3, QP2 and SJ1 had significant coccidiostatic efficacies (inhibition of sporulation) higher than 70%, while isolates FR1, QP2 and QP1 had coccidicidal efficacies (destruction of the oocysts) of 22%, 14% and 8%, respectively, after 14 days of incubation, being a gradual and time-dependent process. To our knowledge, this is the first report regarding the isolation of native predatory fungi from avian feces and demonstration of their lytic activity against coccidia.

Infectious SARS-CoV-2 Particles from Rectal Swab Samples from COVID-19 Patients in Brazil.

The main objective of this study was to investigate the dynamic of SARS-CoV-2 viral excretion in rectal swab (RS), saliva, and nasopharyngeal swab (NS) samples from symptomatic patients and asymptomatic contacts. In addition, in order to evaluate the replication potential of SARS-CoV-2 in the gastrointestinal (GI) tract and the excretion of infectious SARS-CoV-2 from feces, we investigated the presence of subgenomic nucleoprotein gene (N) mRNA (sgN) in RS samples and cytopathic effects in Vero cell culture. A prospective cohort study was performed to collect samples from symptomatic patients and contacts in Rio de Janeiro, Brazil, from May to October 2020. One hundred and seventy-six patients had samples collected at home visits and/or during the follow up, resulting in a total of 1633 RS, saliva, or NS samples. SARS-CoV-2 RNA was detected in 130 (73.9%) patients who had at least one sample that tested positive for SARS-CoV-2. The presence of replicating SARS-CoV-2 in RS samples, measured by the detection of sgN mRNA, was successfully achieved in 19.4% (6/31) of samples, whilst infectious SARS-CoV-2, measured by the generation of cytopathic effects in cell culture, was identified in only one RS sample. Although rare, our results demonstrated the replication capacity of SARS-CoV-2 in the GI tract, and infectious viruses in one RS sample. There is still a gap in the knowledge regarding SARS-CoV-2 fecal-oral transmission. Additional studies are warranted to investigate fecal or wastewater exposure as a risk factor for transmission in human populations.

Creatinine recovery from bovine urine under the effect of different times and temperatures of storage.

Creatinine is a urinary marker used widely in ruminant's experimental trials. However, despite its great importance no data were found in the literature about the best way to store bovine urine samples. In the sheep urine, was observed an increase in the urinary concentration of creatinine when it was stored acidified (pH 2.5 to 3.5) at a temperature of 28 to 39 °C for 150 days of storage. Nevertheless, urine should be stored acidified (pH below 3) to avoid purine derivative degradation, So, aimed to evaluate creatinine recovery in bovine urine as a function of storage time and temperature. A total of 25 animals' urine (10 Nellore cattle and 15 Holstein cattle) were collected. The urine (40 mL) was diluted in 160 mL of distilled water and its pH was corrected to a value lower than 3 using sulfuric acid drops. A sample of the diluted urine was analyzed to obtain the creatinine concentration reference value on the collection day. The remaining urine was fractionated and preserved at room temperature, cooled (4 °C) or frozen (-20 °C and -40 °C). In the urine of five Holstein cattle was added creatine solutions (20, 40 and 60 mg/dL) to evaluate the creatine to creatinine conservation. These urine samples were analyzed on different days after collection (1, 3, 7, 15, 30 and 45 days). The urine without any added creatine was analyzed on Days 1, 3, 7, 10, 15, 30, 45, 60, 90, 120, and 150 of storage. The addition of creatine in the urine caused an increase in the creatinine concentration (P < 0.05) after 30 days of storage at room temperature and under refrigeration (4 °C). In frozen samples, there was no change in creatinine concentration (P > 0.05). However, creatinine recovery was constant (P > 0.05) until day 15 of storage, regardless of the temperature used, when creatine was not added. After 30 days of storage, an effect of time and/or temperature was observed on creatinine recovery (P < 0.05). Urine samples can be stored at any temperature for up to 15 days after collection to estimate the creatinine concentration. Samples that need storage times longer than 15 days should be frozen (at -20 °C and -40 °C) to avoid creatinine concentration variation.

Impact of the insertion of the clinical pharmacist in the Allogeneic Hematopoietic Stem Cells Transplantation team.

INTRODUCTION: Allogeneic Hematopoietic Stem Cells Transplantation (allo-HSCT) is capable of curing patients with neoplastic or non-neoplastic hematologic disorders or of prolonging their survival. This study assessed if the insertion of the clinical pharmacist in the allo-HSCT team modified the outcomes: transplantation-related mortality, grafting failure, incidence of Graft versus Host Disease, hospitalization time, time for grafting, number of readmissions, number of drug-related problems (DRPs), adherence and knowledge about pharmacotherapy. METHODS: Interventional study with historical control carried out in an allo-HSCT unit, in which the intervention group (IG) included 33 individuals who received pharmacotherapy follow-up. Control Group (CG) consisted of 28 individuals. RESULTS: A total of 250 DRPs were identified, 59 team's doubts were clarified, and 309 interventions were conducted in the IG. The DRPs mainly arose from safety (51.60%) and effectiveness (38.40%) problems. A mean of 9.36 (SD = 6.97) interventions per patient was obtained, mainly including dose reductions (19.09%), adjustments in administration time (18.12%), educational activities (15.21%) and drug removal (10.68%). Clinical significance of the interventions was considered high (75.7% extremely significant, very significant or significant), as well as their acceptability (89.7% accepted). Each patient attended a mean of 4.68 pharmaceutical consultations (SD = 1.91) after hospital discharge, presenting increase in knowledge (p = 0.0001) and in adherence (p = 0.0115). There was no evidence of differences between the groups in the other outcomes analyzed. CONCLUSIONS: The pharmacotherapy follow-up allowed detecting several DRPs and performing interventions of high clinical relevance and acceptability, in addition to improving adherence and individualizing the pharmacotherapy.

Can pharmacotherapeutic follow-up after allogeneic hematopoietic stem cell transplantation improve medication compliance?

BACKGROUND: Allogeneic Hematopoietic Stem Cell Transplantation (allo-HSCT) is currently one of the most effective therapies in onco-hematology. For the treatment of the disease and prevention of such complications, a complex pharmacotherapeutic regimen is employed. Non-compliance is prevalent among adolescents and young adults with chronic hematological diseases, being reported by up to 50% of the patients. OBJECTIVE: To evaluate the results of pharmacotherapeutic follow-up on medication compliance and on the knowledge about pharmacotherapy of patients who underwent allo-HSCT. METHODS: A single-arm, open-label and non-randomized intervention study developed in an allo-HSCT outpatient clinic. The participants attended pharmaceutical consultations and had their knowledge about pharmacotherapy and medication compliance measured by MedTake and Brief Medication Questionnaire (BMQ), respectively. RESULTS: A total of 27 patients attended pharmaceutical consultations (4.81 consultations/patient; SD = 1.80). There was an improvement in medication compliance and in knowledge between the first and last consultations (p < 0.05). In the final consultation, 70.37% of the patients showed compliance, with a knowledge rate of 98.35% (SD = 3.63). Non-compliant individuals presented a greater tendency to hospital readmissions. There was no relationship between medication compliance and sociodemographic variables, graft-versus-host disease, and knowledge about pharmacotherapy. CONCLUSIONS: Pharmacotherapeutic follow-up contributed to improving medication compliance. Knowledge about pharmacotherapy alone does not translate into behaviors, which corroborates the complexity of the biopsychosocial factors associated with medication compliance.

Contribution of Pharmaceutical Care to Person-centered Health Care and the Safety of Pharmacotherapy for Hospitalized Older Individuals in Brazil: An Investigative Single-arm Intervention Trial.

BACKGROUND: Adverse drug events (ADE) and medication errors (ME) provide large numbers of victims. Older people are more susceptible to these events, due to the continuing search for several chronic degenerative disease treatments. The Third Global Patient Safety Challenge announced the objective of reducing unnecessary polypharmacy, encouraging deprescription, and aiming to ensure the prescription of medications in an appropriate manner, based on the best evidence and taking into account the individual factors of people. OBJECTIVE: To evaluate whether Pharmaceutical Care (PC), when inserted in a geriatric ward and the context of person-centered health care, cooperates with the safety of pharmacotherapy in older individuals in Brazil. METHODS: This is an investigative, single-arm, preliminary study. INCLUSION CRITERIA: individuals aged ≥60 years and admitted to the geriatric ward between August 2019 to January 2020. The PC (with the practice of pharmacotherapeutic follow-up, medication reconciliation, and pharmacotherapy review) was made available to identify ADE and ME, as well as the associated factors and clinical outcomes, were analyzed. RESULTS: 60 participants were included. It was found that, on hospital admission, 93.3% of them were polymedicated and 86.7% had a history of using potentially inappropriate medications (PIM). ADE and ME were detected in 43 individuals (71.7%) and, in total, 115 incidents were identified, with drugs that act on the nervous system associated with them (31.9%). Acceptance of the PC's recommendations reached the rate of 85.2%. Polypharmacy (p=0.03) and the presence of multiple diseases (p=0.03) had an effect on the presentation of ADE and ME. The number of medications in use decreased in the comparison between admission and hospital discharge (p<0.0001). CONCLUSION: This investigative study indicated that ADE and ME are linked to the polypharmacy in use at the beginning of hospitalization. On the other hand, we showed that the PC (inserted in the multidisciplinary team) contributed to the deprescribing of medications at hospital discharge. Therefore, the PC can provide improvements in this scenario.

The SISBIOTA-Diptera Brazilian Network: A long-term survey of Diptera from unexplored Brazilian Western Arc of Amazon, Cerrado, and Pantanal

ABSTRACT The SISBIOTA-BRASIL was a three-year multimillion-dollar research program of the Brazilian government to document plants and animals in endangered/understudied areas and biomes in Brazil. Distributional patterns and the historical events that generated them are extensively unknown regarding Brazilian fauna and flora. This deficiency hinders the development of conservation policies and the understanding of evolutionary processes. Conservation decisions depend on precise knowledge of the taxonomy and geographic distribution of species. Given such a premise, we proposed to research the diversity of Diptera of the Brazilian western arc of Amazon, Cerrado, and Pantanal in the states of Mato Grosso, Mato Grosso do Sul, and Rondônia. Three important biomes of the South American continent characterize these Brazilian states: Amazon forest, Cerrado (Brazilian Savannah), and Pantanal. Besides their ecological relevance, these biomes historically lack intensive entomological surveys. Therefore, they are much underrepresented in the Brazilian natural history collections and in the scientific literature, which is further aggravated by the fact that these areas are being exponentially and rapidly converted to commercial lands. Our project involved over 90 collaborators from 24 different Brazilian institutions and one from Colombia among researchers, postdocs, graduate and undergraduate students, and technicians. We processed and analyzed nearly 300,000 specimens from ~60 families of Diptera collected with a large variety of methods in the sampled areas. Here, we provide a detailed overview of the genera and species diversity of 41 families treated. Our results point to a total of 2,130 species and 514 genera compiled and identified for the three states altogether, with an increase of 41% and 29% in the numbers of species and genera known for the three states combined, respectively. Overall, the 10 most species-rich families were Tachinidae, Cecidomyiidae, Tabanidae, Psychodidae, Sarcophagidae, Stratiomyidae, Bombyliidae, Syrphidae, Tephritidae, and Asilidae. The 10 most diverse in the number of genera were Tachinidae, Stratiomyidae, Asilidae, Mycetophilidae, Syrphidae, Tabanidae, Muscidae, Dolichopodidae, Sarcophagidae, and Chloropidae. So far, 111 scientific papers were published regarding taxonomic, phylogenetic, and biogeographical aspects of the studied families, with the description of 101 new species and three new genera. We expect that additional publications will result from this investigation because several specimens are now curated and being researched by specialists.

Selection dynamics of HCV genotype 3 resistance-associated substitutions under direct-acting antiviral therapy pressure.

The chronic hepatitis C (CHC) treatment is currently based on the use of direct-acting antivirals (DAAs), and patients infected with hepatitis C virus genotype 3 (GT3) have emerged as a more difficult-to-cure population. The NS5A inhibitor daclatasvir (DCV) and sofosbuvir (SOF), an NS5B viral polymerase inhibitor, are among the drugs that compose more effective and safer treatment regimens. The virus genetic variability is related to resistance-associated substitutions (RASs) that adversely impact DAAs effectiveness. The aims of this study were to analyze the association of NS5A and NS5B RASs and other clinical factors with DAAs regimens effectiveness in patients with GT3 CHC infection. This was a prospective cohort study performed in a Brazilian university hospital. Individuals older than 18 years with GT3 CHC treated with SOF + DCV ± ribavirin (RBV) or SOF + peginterferon (PEG) + RBV were included. Blood samples were collected at baseline and post-treatment. A total of 121 patients were included. Sustained virological response rates were 87.6% for the SOF + DCV ± RBV group and 80.0% for the SOF + PEG + RBV arm. Cirrhosis, prior treatment with interferon/PEG + RBV, and baseline NS5A RAS were associated with higher risk of treatment failure. The NS5A analysis suggested that A30K, Y93H, and RAS at site 62 were related to failure. Interestingly, a likely compensatory effect was shown between A30K and A62T. Emergence of Y93H was always associated with RAS at position 62. The RASs dynamics comprehension is an important tool to indicate more effective treatment for GT3 patients.

[Central nervous system adverse events potentially associated with drugs used for COVID-19: scoping reviewEventos adversos en el sistema nervioso central potencialmente relacionados con los medicamentos para tratar la COVID-19: revisión exploratoria]. / Eventos adversos no sistema nervoso central potencialmente relacionados aos medicamentos utilizados na COVID-19: revisão de escopo.

Objective: To identify central nervous system (CNS) adverse events potentially associated with prophylaxis or drug treatment for COVID-19, and to describe the characteristic of the individuals affected. Methods: A scoping review was performed using a search strategy to retrieve articles from PubMed, EMBASE, SciELO, Scopus, CINAHL and BVS databases. Studies reporting on individuals receiving prophylactic or curative drugs for COVID-19 with at least one CNS adverse event were included. Articles reporting on CNS adverse events associated with medication for other health conditions were excluded. Results: The search retrieved 1 547 articles, eight of which met the inclusion criteria. Seven studies had an observational design. A total of 3 035 individuals were assessed, of whom 1 701 were health care professionals and 1 978 were women. Curative treatment with hydroxychloroquine, chloroquine, lopinavir/ritonavir, and azithromycin was the most frequent (n = 5). The most common adverse events were headache, dizziness, mood disturbances, and drowsiness. Suicide was the most frequent severe event. Six adverse events were unexpected for hydroxychloroquine, chloroquine, and doxycycline. Conclusion: Potential CNS adverse events were unspecific and in general potentially associated with the use of hydroxychloroquine (monotherapy or associated with antibiotics). The data confirm the unfavorable risk/benefit profile of these drugs for the prevention and management of signs and symptoms of SARS-CoV-2 infection.

Objetivo: Identificar los eventos adversos en el sistema nervioso central (SNC) potencialmente relacionados con el uso de medicamentos empleados para profilaxis o tratamiento de la COVID-19, y caracterizar a las personas afectadas. Métodos: Se realizó una revisión exploratoria a partir de una estrategia de búsqueda en las bases de datos PubMed, EMBASE, SciELO, Scopus, Cummulative Index to Nursing and Allied Health Literature (CINAHL) y la Biblioteca Virtual de Salud (BVS). Se incluyeron estudios de personas que emplearon medicamentos con fines profilácticos o curativos para la COVID-19 y presentaron al menos un evento adverso en el SNC. Se excluyeron los artículos en los cuales se notificaron eventos adversos en el SNC potencialmente relacionados con medicamentos para tratar otros problemas de salud. Resultados: Se recuperaron 1 547 artículos, de los cuales ocho cumplieron con los criterios de admisibilidad. Siete estudios tuvieron un diseño observacional. Se analizaron 3 035 personas, de las cuales 1 701 eran profesionales de salud y 1 978, mujeres. El tratamiento más utilizado fue el curativo (n = 5), con hidroxicloroquina, cloroquina, lopinavir/ritonavir y azitromicina. Los eventos adversos comúnmente citados fueron dolor de cabeza, mareos, trastornos del estado de ánimo y somnolencia. El suicidio fue el evento grave más frecuente. Seis eventos inesperados (mioclonías, temblor, trastorno de la marcha, disgeusia, hiperhidrosis y desasosiego) guardaron relación con el empleo de hidroxicloroquina, cloroquina y doxiciclina. Conclusión: Los eventos adversos del SNC fueron inespecíficos y, en general, posiblemente estuvieron relacionados con el uso de hidroxicloroquina (sola o combinada) para el tratamiento curativo de la COVID-19. Los datos corroboran la relación desfavorable de riesgo/beneficio de esos medicamentos en la prevención y el manejo de los signos y síntomas de la infección por el SARS-CoV-2.

Depletion of Na+/H+ Exchanger Isoform 1 Increases the Host Cell Resistance to Trypanosoma cruzi Invasion.

Na+/H+ exchanger isoform 1 (NHE1), a member of a large family of integral membrane proteins, plays a role in regulating the cortical actin cytoskeleton. Trypanosoma cruzi, the agent of Chagas disease, depends on F-actin rearrangement and lysosome mobilization to invade host cells. To determine the involvement of NHE1 in T. cruzi metacyclic trypomastigote (MT) internalization, the effect of treatment in cells with NHE1 inhibitor amiloride or of NHE1 depletion was examined in human epithelial cells. MT invasion decreased in amiloride-treated and NHE1-depleted cells. The phosphorylation profile of diverse protein kinases, whose activation is associated with remodeling of actin fibers, was analyzed in amiloride-treated and NHE1-depleted cells. In amiloride-treated cells, the phosphorylation levels of protein kinase C (PKC), focal adhesion kinase (FAK) and Akt were similar to those of untreated cells, whereas those of extracellular signal-regulated protein kinases (ERK1/2) increased. In NHE1-deficient cells, with marked alteration in the actin cytoskeleton architecture and in lysosome distribution, the levels of phospho-PKC and phospho-FAK decreased, whereas those of phospho-Akt and phospho-ERK1/2 increased. These data indicate that NHE1 plays a role in MT invasion, by maintaining the activation status of diverse protein kinases in check and preventing the inappropriate F-actin arrangement that affects lysosome distribution.

Gp35/50 mucin molecules of Trypanosoma cruzi metacyclic forms that mediate host cell invasion interact with annexin A2.

Host cell invasion is a critical step for infection by Trypanosoma cruzi, the agent of Chagas disease. In natural infection, T. cruzi metacyclic trypomastigote (MT) forms establish the first interaction with host cells. The gp35/50 mucin molecules expressed in MT have been implicated in cell invasion process, but the mechanisms involved are not well understood. We performed a series of experiments to elucidate the mode of gp35/50-mediated MT internalization. Comparing two parasite strains from genetically divergent groups, G strain (TcI) and CL strain (TcVI), expressing variant forms of mucins, we demonstrated that G strain mucins participate in MT invasion. Only G strain-derived mucins bound to HeLa cells in a receptor-dependent manner and significantly inhibited G strain MT invasion. CL strain MT internalization was not affected by mucins from either strain. HeLa cell invasion by G strain MT was associated with actin recruitment and did not rely on lysosome mobilization. To examine the involvement of annexin A2, which plays a role in actin dynamic, annexin A2-depleted HeLa cells were generated. Annexin A2-deficient cell lines were significantly more resistant than wild type controls to G strain MT invasion. In a co-immunoprecipitation assay, to check whether annexin A2 might be the receptor for mucins, protein A/G magnetic beads crosslinked with monoclonal antibody to G strain mucins were incubated with detergent extracts of MT and HeLa cells. Binding of gp35/50 mucins to annexin A2 was detected. Both G strain MT and purified mucins induced focal adhesion kinase activation in HeLa cells. By confocal immunofluorescence microscopy, colocalization of invading G strain MT with clathrin was visualized. Inhibition of clathrin-coated vesicle formation reduced parasite internalization. Taken together, our data indicate that gp35/50-mediated MT invasion is accomplished through interaction with host cell annexin A2 and clathrin-dependent endocytosis.

A Standard Procedure for In Vitro Digestion Using Rumen Fermenters: A Collaborative Study.

Our objective was to propose and evaluate a standard procedure for the evaluation of in vitro dry matter digestibility for ruminant feeds, using artificial fermenters. A collaborative study was performed with seven feed analysis laboratories and four feeds (Tifton 85 hay, corn silage, soybean hulls, and soybean meal). Two types of artificial fermenters were evaluated (DaisyII Ankom and TE-150 Tecnal). Each laboratory received 80 sealed filter bags with samples (20 per feed), eight blank filter bags, a plastic bag with buffer solution reagents, and instructions describing how to conduct a 48 h in vitro assay using an artificial fermenter and how to collect bovine ruminal inoculum. On average, the contribution of laboratory effect to the total random variance was 24%, being less than the contribution of equipment (42%) and error (34%). The repeatability ranged from 3.34 to 5.79%, across feeds. The reproducibility ranged from 5.93 to 8.94% across feeds, which implied Horwitz ratios ranging from 2.94 to 4.10. Due to the specific characteristics of the analytical entity evaluated here, which is defined by the method itself, the proposed method was considered reproducible. The results highlighted that, if the method is followed exactly, its results are precise and present adequate levels of repeatability and reproducibility.

Protein requirements of hair sheep raised in warm areas: a multi-study approach.

The protein requirements of hair sheep are a key factor in increasing profitability and decreasing the environmental impacts of nitrogen excretion. The objective of this study was to evaluate the protein requirements of hair sheep. A database with 382 individual records (269 intact and 113 castrated males) comprising information from 11 studies was used. The studies provided different levels of metabolisable protein intake (MPI) and of N retention, allowing the development of equations to predict the net protein (NP) and the metabolisable protein (MP) requirements. The efficiency of MP use for gain (kpg) was calculated using the equation of daily protein retained against daily MPI above maintenance. The efficiency of MP use for maintenance (kpm) was computed as the ratio between the NP for maintenance (NPm) and the MP for maintenance (MPm). The NPm (1.32 g/kg0.75 EBW) did not differ between sex. The kpm was 0.34, and the kpg was 0.25. The MPm estimated was 3.21 g/kg0.75 BW. Sex affects the protein requirements for gain (NPg). The protein requirements of hair sheep differ from those recommended by feeding systems for sheep. The equations provided herein may improve the optimisation of protein nutrition of sheep, thereby minimising the environmental impacts of sheep production.

Protocol for the implementation of pharmaceutical care in geriatrics: strategy for safety in health care.

OBJECTIVE: To describe the experience of the implementation of pharmaceutical care in a geriatric hospital unit and to propose an instructional protocol for the practice. METHODS: Experience report that became the practice manual of pharmaceutical care in geriatrics (MaP-CFarmaGeri) of a Brazilian hospital and was structured in three topics (1. Situational diagnosis; 2. Adequacy of the procedure and service provision; 3. Practice exercise). RESULTS: The situational diagnosis comprised the collection of data on the structure of the ward and the epidemiological profile. The pharmaceutical services provided included pharmacotherapeutic follow-up, medication reconciliation and pharmacotherapy review. The certification of the content of this procedure was attested by specialists from a multiprofessional team and the technique served more than 60 patients in practice, with good acceptance by the participants. FINAL CONSIDERATIONS: The MaP-CFarmaGeri proved to be a satisfactory strategy in the implementation of pharmaceutical care in geriatrics and can support this insertion in similar locations.

Antioxidant and anti-inflammatory effects of fractions from ripe fruits of Solanum lycocarpum St. Hil. (Solanaceae) and putative identification of bioactive compounds by GC-MS and LC-DAD-MS.

Brazilian biodiversity includes species of the genus Solanum that have several biological activities, in addition to their relevance to agriculture, economics and popular medicine. The ripe fruits of Solanum lycocapum are an important nutritional food source, since they have levels of vitamin C, total soluble sugars, sucrose, phosphorus, and iron comparable or exceed the levels present in fruits such as pineapples, bananas, oranges, and mangoes. The pulp of the fruit is consumed by the population, and it is also used to produce jellies; to make marmalade, replacing the quince, and it can also be mixed with peaches in the preparation of peach. The objective of this study was to evaluate the anti-inflammatory and antioxidant activities of fractions obtained from the ripe fruits and to identify the constituents with these biological properties. The ripe fruits were collected, dried, crushed, and subjected to extraction by exhaustive percolation, obtaining an ethanol extract that was partitioned with solvents of increasing polarities, obtaining hexane (HEX), ethyl acetate (AC), and hydroethanol (HE) fractions. The AC fraction showed higher antioxidant potential compared to BHT (2,6-di-tert-butyl-4-methylphenol) and similar activity to AA (ascorbic acid) by DPPH (1,1-diphenyl-2-picrylhydrazyl) radical assay, while HEX and HE fractions exhibited of IC50 values similar to BHT. The AC fraction also presented similar activity to BHT by FRAP (Ferric Reducing Antioxidant Power) test. Intraperitoneal treatment with HEX (100 and 300 mg/kg) and HE (100 mg/kg) fractions caused significant inhibition of paw edema induced by carrageenan, 4 and 6 h after the inflammatory stimuli. When analyzed by GC-MS, fatty acids, phytosterols, and triterpenoid were identified in the HEX fraction, whilst 31 compounds were annotated in the AC and HE fractions analyzed by LC-DAD-MS, being phenylpropanoid derivatives, chlorogenic acids, and steroidal glycoalkaloids. The ripe fruits of S. lycocarpum have antioxidant and anti-inflammatory activities, and the detected chemical compounds, especially caffeoylquinic acid derivatives, spermidine, stigmasterol, and ß-sitosterol, may be correlated with these activities. The ripe fruits of this species can be a food alternative rich in bioactive compounds and with benefits for human health.

A Case Report of a Rare Pediatric Brain Tumor: Congenital Glioblastoma.

Congenital brain tumors are extremely rare; even with advances in prenatal imaging tests, it is still uncommon. Congenital glioblastoma (GBM) is a rare childhood tumor. With less than 50 cases described in the literature, it corresponds to less than 2% of tumors in children under two months of life. Moreover, it has a markedly poor prognosis due to the risk of intracranial hemorrhage, especially during surgical resection. This study reports the case of a 20-day-old asymptomatic child who presented with increased head circumference during a pediatric routine check-up. A transfontanellar ultrasound was performed, exhibiting hydrocephalus, large parenchymal hemorrhage, and expansive formation. Magnetic resonance imaging pointed to a massive infiltrative lesion, with heterogeneous enhancement, delimiting central areas of necrosis with hematic material inside, associated with a compressive effect on the adjacent parenchyma. Additional histopathological analysis, immunohistochemistry, and DNA methylation test confirmed the diagnosis of GBM. The patient was submitted to surgical intervention and chemotherapy, achieving a 26-month-old survival by the time this study was written.