Guide catheter extension use are associated with higher procedural success in chronic total occlusion percutaneous coronary interventions.

BACKGROUND: Guide catheter extensions (GCEs) increase support and facilitate equipment delivery, but aggressive instrumentation may be associated with a higher risk of complications. AIM: Our aim was to assess the impact of GCEs on procedural success and complications in patients submitted to chronic total occlusion (CTO) percutaneous coronary intervention (PCI). METHODS: We analyzed data from the multicenter LATAM CTO Registry. Procedural success was defined as <30% residual stenosis and TIMI 3 distal flow. Major adverse cardiac and cerebrovascular events (MACCE) was defined as the composite of all-cause death, myocardial infarction, target vessel revascularization, and stroke. Propensity score matching (PSM) was used to compare outcomes with and without GCE use. RESULTS: From August 2010 to August 2021, 3049 patients were included. GCEs were used in 438 patients (14.5%). In unadjusted analysis, patients in the GCE group were older and had more comorbidities. The median J-CTO score and its components were higher in the GCE group. After PSM, procedural success was higher with GCE use (87.7% vs. 80.5%, p = 0.007). The incidence of coronary perforation (odds ratio [OR]: 1.46, 95% confidence interval [CI]: 0.78-2.71, p = 0.230), bleeding (OR: 1.99, 95% CI: 0.41-2.41, p = 0.986), in-hospital death (OR: 1.39, 95% CI: 0.54-3.62, p = 0.495) and MACCE (OR: 1.07, 95% CI: 0.52-2.19, p = 0.850) were similar in both groups. CONCLUSION: In a contemporary, multicenter cohort of patients undergoing CTO PCI, GCEs were used in older patients, with more comorbidities and complex anatomy. After PSM, GCE use was associated with higher procedural success, and similar incidence of adverse outcomes.

Pathological findings and their association with diseases of captive psittacine birds native to Mexico.

The captivity and use of native psittacine birds is prohibited in Mexico. However, as these birds are among the groups most affected by illegal trafficking, they are commonly found as companion animals. Nevertheless, it is difficult to obtain information on their health. Therefore, a retrospective study was conducted of the clinical histories and necropsy reports of native psittacines that had been submitted to the Bird Disease Diagnostic and Research Laboratory of the Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, from 2006 to 2017. The lesions were classified according to type and anatomical location and the diseases were classified as infectious or non-infectious. During this period, 252 psittacines were submitted, the most frequent of which were the red-lored parrot (Amazona autumnalis), orange-fronted parakeet (Eupsittula canicularis) and scarlet macaw (Ara macao). The lesions were primarily located in the digestive and respiratory systems. By integrating the clinical histories and post-mortem findings, we concluded that nutritional disorders were the most frequent non-infectious diseases, systemic bacterial infections were the most frequent infectious conditions, the primary parasite was Sarcocystis spp and the most frequent neoplasm was multicentric lymphoma.

Dispharynx nasuta (Nematoda: Acuarioidea: Acuariidae) infestation in exotic red-rumped parrots (Psephotus haematonotus) in captivity in the State of Michoacan, Mexico.

Dispharynx nasuta is a widespread nematode parasite located in the proventriculus. This parasite may cause mortality in free-living birds or in captivity. However, reports of this parasite in psittacines are scarce. In a private aviary, in Zamora, Michoacan, Mexico six red-rumped parrots (Psephotus haematonotus) died in one month, from mid-August to mid-September 2016, and one more specimen died during the examination. Prior to death, the birds presented depression, ruffled feathers, crop atony, and regurgitation. Upon necropsy, ulcers in the proventriculus, and hemorrhagic content associated with the presence of round worms was observed. At histopathology examination, the parasites were identified as Spirurida nematodes, located in the proventricular and ventricular mucosa. Specimens were submitted to the parasitology laboratory of the College of Veterinary Medicine and Husbandry, National Autonomous University of Mexico (UNAM), where the parasites were identified as D. nasuta. This is the first report of D. nasuta in P. haematonotus and the first report of D. nasuta in Mexico.

Immunotherapy With Egg Yolk Eimeria sp.-Specific Immunoglobulins in SPF Leghorn Chicks Elicits Successful Protection Against Eimeria tenella Infection.

Avian coccidiosis is the first to most economically important parasite disease affecting poultry industries worldwide. Current prevention measures are largely based upon prophylactic chemotherapy supplemented by the application of live attenuated or wild-type parasite vaccines. However, the rising appearance of drug resistance, consumer's concern for antibiotics use in poultry production and higher manufacturing cost of live vaccines has driven to adopt new technologies aimed at increasing animal health and production efficiency. Supplementing chickens with egg yolk Eimeria sp.-specific immunoglobulins can be a viable alternative to avoid severe outbreaks of the disease. Twelve-week-old SPF White Leghorn chickens were experimentally infected with a large dose of E. tenella. During the prepatent period, the birds were supplemented by oral gavage with 60 or 120 mg/bird of hyperimmune egg yolk Eimeria species-specific immunoglobulins Y (Supracox®, SC) on a daily basis. The animals were euthanized 7 days post-infection (PI) and their passive immune protection was evaluated. Birds treated with 120 mg/bird of SC showed more viability, increased body weight gain (BWG), a normal hematocrit level (HCT), reduced oocyst output per gram of feces (OPG) or cecal tissue (OPGC), and fewer cecal lesions compared to the untreated infected (UI) control group. Birds supplemented with 60 mg/bird of SC did not show any significant difference on BWG, HCT, OPG, OPGC, and cecal lesion score when compared with the UI group. An ELISA test of the SC showed a weak cross-reactivity of IgY toward two asexual zoite stages of E. tenella. Western blot analysis of the sporozoite with SC showed few antigens barely recognized, while more stained bands were detected in the merozoite (≈82, ≈60, ≈54, ≈40, ≈38, ≈27.5, and ≈13 kDa). Oral immunotherapy using egg yolk polyclonal IgYs against Eimeria sp. represents an effective and natural resource against severe E. tenella infection favoring the gradual withdrawal of the anticoccidial drugs and antibiotics.

Opuntia ficus-indica Extract and Isorhamnetin-3-O-Glucosyl-Rhamnoside Diminish Tumor Growth of Colon Cancer Cells Xenografted in Immune-Suppressed Mice through the Activation of Apoptosis Intrinsic Pathway.

This study aimed to evaluate the effects of Opuntia ficus-indica extract (OFI-E) and its glycoside isorhamnetin-3-O-glucosyl-rhamnoside (IGR) on the growth of human colorectal adenocarcinoma cells and in a xenografted-immunosuppressed mice model. The IC50 values of OFI-E and IGR on colon cancer cells (HT-29 RFP) were determinate, as well as their effects on the cell cycle and apoptosis induction. OFI-E and IGR produced an increased in apoptosis induction, ROS production and a G0/G1 cell cycle arrest. In xenografted-inmunosupressed mice, OFI-E and IGR reduced the tumor growth rate, myeloperoxidase activity and total cholesterol levels. OFI-E and IGR reduced the tumor growth through the overexpression of cleaved Caspase-9, Hdac11, and Bai1 proteins. OFI-E reduced the expression of bcl-2. Results demonstrated the chemopreventive effects of OFI-E, and its purified compound IGR, showing their potential as an alternative in the treatment of colorectal cancer.

Female ejaculation: An update on anatomy, history, and controversies.

Female ejaculation is a contentious topic. From a review of the literature, history indicates that it is not a modern concept; some females were aware of it in times past without understanding the role of the fluid or composition of the ejaculate. Over time, scholars experimented, mainly with anatomical studies, in an attempt to identify the source of the ejaculate and explore its physiological and anatomical benefits for the female sexual experience. Despite these studies, views about female ejaculation remain controversial and inconsistent, with no clear conclusion as to its function. This review discusses the history of studies of female ejaculation and presents various hypotheses from an anatomical and physiological perspective. After reviewing 44 publications from 1889 to 2019, it became apparent that clinical and anatomical studies conducted during recent decades provide substantial evidence in support of the female ejaculatory phenomenon. Anatomical studies have shown that the ejaculate originates in the paraurethral (Skene's) glands, but its composition has been debated. Female ejaculate differs from urine in its creatinine and urea concentrations. The fluid also contains prostate specific antigen (PSA) and could have antibacterial properties that serve to protect the urethra. While the specific function of female ejaculation remains a topic of debate, there is sufficient evidence to support the existence of the phenomenon.

First report of Boydaia falconis (Trombidiformes: Ereynetidae: Speleognathinae) in a Merlin (Falco columbarius) (Falconiformes: Falconidae) in Veracruz, Mexico.

The report presented herein documents the finding of mites in the nares of a Merlin (Falco columbarius) (Linnaeus, 1758; Falconiformes: Falconidae) during its capture for identification and ringing at the conservation reserve area in the municipality of Cansaburro, state of Veracruz, Mexico.The mites were collected from the nostril of the bird and identified as Boydaia falconis (Fain, 1956; Trombidiformes: Ereynetidae: Speleognathinae). There are few records of nasal mites in Faconiforms in North America. This is the first report of Boydaia falconis in falconiform hosts from Mexico. Further study is required on these mites to aid in our understanding of the biology, ecology and symbiotic relationships of speleognathine nasal mites.

Chronic total occlusion percutaneous coronary intervention in Latin America.

OBJECTIVES: To report clinical, angiographic characteristics, outcomes, and predictors of unsuccessful procedures in patients who underwent chronic total occlusion (CTO) percutaneous coronary interventions (PCI) in Latin America. BACKGROUND: CTO PCI has been increasingly performed worldwide, but there is a lack of information in this region. METHODS: An international multicenter registry was developed to collect data on CTO PCI performed in centers in Latin America. Patient, angiographic, procedural and outcome data were evaluated. Predictors of unsuccessful procedures were assessed by multivariable analysis. RESULTS: We have included data related to 1,040 CTO PCIs performed in seven countries in Latin America (Argentina, Brazil, Chile, Colombia, Ecuador, Mexico, and Puerto Rico). The mean age was 64 ± 10 years, and CTO PCI was performed mainly for angina control (81%) or treatment of a large ischemic area (30%). Overall technical success rate was 82.5%, and it was achieved with antegrade wire escalation in 81%, antegrade dissection/re-entry in 8% and with retrograde techniques in 11% of the successful procedures. Multivariable analysis identified moderate/severe calcification, a blunt proximal cap and a previous attempt as independent predictors of unsuccessful procedures. In-hospital major adverse cardiovascular events (MACE) occurred in 3.1% of the cases, death in 1% and cardiac tamponade in 0.9% CONCLUSIONS: CTO PCI in Latin America has been performed mainly for ischemia relief. Procedures were associated with a success rate above 80% and low incidence of MACE. Predictors of unsuccessful procedures were similar to those previously reported in the literature.

Fat-Soluble Vitamin Supplementation Levels in Diets for Laying Hens from 28 to 44 Weeks of Age

This study aimed to examine the impact of levels of dietary supplementation with fat-soluble vitamins on the production performance and egg quality of laying hens. Three hundred Hy-Line White W-36 laying hens were evaluated from 28 to 44 weeks of age. The birds were allotted to one of six treatments in a randomized block design with 10 replicates with five birds each. Performance and egg quality parameters were evaluated in four 28-day periods. A corn and soybean meal-based basal diet was formulated so as to meet the nutritional requirements of the animals, with the exception of fat-soluble vitamins. The treatments consisted of dietary supplementation with 0%, 33.3%, 66.7%, 100.0%, 133.3% or 166.7% of fat-soluble vitamins (100% supplementation consisted of 7500 IU, 2000 IU, 10 IU and 1.8 mg of vitamins A, D3, E and K per kilogram of diet, respectively). Eggshell weight, shell thickness, shell strength, feed intake, egg weight, feed conversion per egg mass and feed conversion per dozen eggs showed a quadratic response (p0.05) to the treatments, whereas egg mass responded linearly. Optimal results were obtained at an average fat-soluble vitamin supplementation level of 109%, which corresponds to 8175 IU of vitamin A, 2180 IU of vitamin D3, 10.9 IU of vitamin E and 1.96 mg of vitamin K per kilogram of diet.(AU)

Fat-Soluble Vitamin Supplementation Levels in Diets for Laying Hens from 28 to 44 Weeks of Age

This study aimed to examine the impact of levels of dietary supplementation with fat-soluble vitamins on the production performance and egg quality of laying hens. Three hundred Hy-Line White W-36 laying hens were evaluated from 28 to 44 weeks of age. The birds were allotted to one of six treatments in a randomized block design with 10 replicates with five birds each. Performance and egg quality parameters were evaluated in four 28-day periods. A corn and soybean meal-based basal diet was formulated so as to meet the nutritional requirements of the animals, with the exception of fat-soluble vitamins. The treatments consisted of dietary supplementation with 0%, 33.3%, 66.7%, 100.0%, 133.3% or 166.7% of fat-soluble vitamins (100% supplementation consisted of 7500 IU, 2000 IU, 10 IU and 1.8 mg of vitamins A, D3, E and K per kilogram of diet, respectively). Eggshell weight, shell thickness, shell strength, feed intake, egg weight, feed conversion per egg mass and feed conversion per dozen eggs showed a quadratic response (p0.05) to the treatments, whereas egg mass responded linearly. Optimal results were obtained at an average fat-soluble vitamin supplementation level of 109%, which corresponds to 8175 IU of vitamin A, 2180 IU of vitamin D3, 10.9 IU of vitamin E and 1.96 mg of vitamin K per kilogram of diet.

[(7-chloroquinolin-4-yl)amino]acetophenones and their copper(II) derivatives: Synthesis, characterization, computational studies and antimalarial activity.

The synthesis of the compounds [(7-chloroquinolin-4-yl)amino]acetophenones (4, 5) and their copper(II) complexes (4a, 5a) is reported. The compounds were characterized using a wide range of spectroscopic and spectrometric techniques, such as FTIR, UV-vis, NMR, EPR, ESI-CID-MS2. The spectral results suggested that the ligand acted as chelating species coordinating the metal through the endocyclic nitrogen of the quinoline ring in both complexes, with general formulae expressed in two ways, according to the phase in which they are: [Cu(L)2Cl2] for solid phase and [Cu(L)2][2Cl] for liquid phase. The EPR study of the Cu (II) complexes indicated a probable distorted tetrahedral coordination geometry. This result was confirmed by the calculated optimized structures at the DFT/B3LYP method with the 6-31G (d,p) basis set. The characterization of the fragmentation pattern of protonated free ligands was extended here to fragments as low as m/z 43, while for coordination complexes it extends to fragments at m/z 80 and m/z 111. The antimalarial activity of the compounds was determined through three different tests: inhibitory activity against in vitro growth of Plasmodium falciparum (W2), inhibition of hemozoin formation (ß-hematin) and in vitro inhibitory activity against recombinant falcipain-2, where compound 5 showed considerable activity. However, the activity of free ligands against P. falciparum was increased by complexing with the Cu (II) metal ion. The values of the HOMO-LUMO energy gap of 3.847 eV (4a) and 3.932 eV (5a) were interpreted with high chemical activity and thus, could influence on biological activity. In both compounds, the total electron density surface mapped with electrostatic potential clearly revealed the presence of high negative charge on the Cu atom. Also, this study reported the molecular docking of free ligands (4, 5) using software package ArgusLab 4.0.1. The results revealed the importance of water molecules as interaction bridges through hydrogen bonds between free ligands and ß-hematin; at the same time, the hypothesis that π-π interaction between quinoline derivatives and the electronic system of hematin governs the formation of adducts was confirmed.

Prevalence of Ehrlichia canis (Rickettsiales: Ehrlichieae) DNA in Tissues From Rhipicephalus sanguineus (Acari: Ixodidae) Ticks in Areas Endemic for Canine Monocytic Ehrlichiosis in Brazil.

Canine monocytic ehrlichiosis (CME) is a disease caused by the obligate intracellular bacterium Ehrlichia canis. Tropical lineages of Rhipicephalus sanguineus ticks play an essential role in the transmission of this pathogen. The aim of the present study was to evaluate the prevalence of E. canis DNA in tissue from R. sanguineus ticks in areas endemic for CME in Brazil and quantify levels of E. canis DNA in dissected tissues from these samples. A total of 720 ticks were collected from 72 dogs (36 dogs from the city Araçatuba in São Paulo state and 36 from Campo Grande in the state of Mato Grosso do Sul). Ticks were dissected to collect the guts, ovaries and salivary gland. A quantitative polymerase chain reaction (qPCR) targeting the disulphide bond formation (dsb) protein gene was performed to quantify the level of E. canis infection. The E. canis dsb-qPCR assay was positive for 31.9, 10, and 15.2% of the gut, ovary, and salivary glands, respectively. The average gut, ovary, and salivary gland bacterial load estimated by qPCR was 1.21 × 103, 2.60 × 103, and 4.92 × 103 gene copies/µl, respectively. This is the first report of E. canis DNA in ovaries of R. sanguineus ticks parasitizing dogs in these CME-endemic areas. These observations raise the possibility of E. canis trans-ovarial transmission.

Antimalarial activity of single-dose DSM265, a novel plasmodium dihydroorotate dehydrogenase inhibitor, in patients with uncomplicated Plasmodium falciparum or Plasmodium vivax malaria infection: a proof-of-concept, open-label, phase 2a study.

BACKGROUND: DSM265 is a novel, long-duration inhibitor of plasmodium dihydroorotate dehydrogenase (DHODH) with excellent selectivity over human DHODH and activity against blood and liver stages of Plasmodium falciparum. This study aimed to assess the efficacy of DSM265 in patients with P falciparum or Plasmodium vivax malaria infection. METHODS: This proof-of-concept, open-label, phase 2a study was conducted at the Asociación Civil Selva Amazónica in Iquitos, Peru. Patients aged 18-70 years, weighing 45-90 kg, who had clinical malaria (P falciparum or P vivax monoinfection) and fever within the previous 24 h were eligible. Exclusion criteria were clinical or laboratory signs of severe malaria, inability to take oral medicine, and use of other antimalarial treatment in the preceding 14 days. Patients were divided into cohorts of those with P falciparum (cohort a) or P vivax (cohort b) infection. Two initial cohorts received single oral doses of 400 mg DSM265. Patients were followed up for efficacy for 28 days and safety for 35 days. Further cohorts received escalated or de-escalated doses of DSM265, after safety and efficacy assessment of the initial dose. The primary endpoints were the proportion of patients achieving PCR-adjusted adequate clinical and parasitological response (ACPR) by day 14 for patients infected with P falciparum and the proportion of patients achieving a crude cure by day 14 for those infected with P vivax. Cohort success, the criteria for dose escalation, was defined as ACPR (P falciparum) or crude cure (P vivax) in at least 80% of patients in the cohort. The primary analysis was done in the intention-to-treat population (ITT) and the per-protocol population, and safety analyses were done in all patients who received the study drug. This study is registered at ClinicalTrials.gov (NCT02123290). FINDINGS: Between Jan 12, 2015, and Dec 2, 2015, 45 Peruvian patients (24 with P falciparum [cohort a] and 21 with P vivax [cohort b] infection) were sequentially enrolled. For patients with P falciparum malaria in the per-protocol population, all 11 (100%) in the 400 mg group and eight (80%) of ten in the 250 mg group achieved ACPR on day 14. In the ITT analysis, 11 (85%) of 13 in the 400 mg group and eight (73%) of 11 in the 250 mg group achieved ACPR at day 14. For the patients with P vivax malaria, the primary endpoint was not met. In the per-protocol analysis, none of four patients who had 400 mg, three (50%) of six who had 600 mg, and one (25%) of four who had 800 mg DSM265 achieved crude cure at day 14. In the ITT analysis, none of five in the 400 mg group, three (33%) of nine in the 600 mg group, and one (14%) of seven in the 800 mg group achieved crude cure at day 14. During the 28-day extended observation of P falciparum patients, a resistance-associated mutation in the gene encoding the DSM265 target DHODH was observed in two of four recurring patients. DSM265 was well tolerated. The most common adverse events were pyrexia (20 [44%] of 45) and headache (18 [40%] of 45), which are both common symptoms of malaria, and no patients had any treatment-related serious adverse events or adverse events leading to study discontinuation. INTERPRETATION: After a single dose of DSM265, P falciparum parasitaemia was rapidly cleared, whereas against P vivax, DSM265 showed less effective clearance kinetics. Its long duration of action provides the potential to prevent recurrence of P falciparum after treatment with a single dose, which should be further assessed in future combination studies. FUNDING: The Global Health Innovative Technology Fund, the Bill & Melinda Gates Foundation, the National Institutes of Health (R01 AI103058), the Wellcome Trust, and the UK Department of International Development.

Identification of Leishmania spp. promastigotes in the intestines, ovaries, and salivary glands of Rhipicephalus sanguineus actively infesting dogs.

Sand flies are recognized as the major vector of canine visceral leishmaniasis. However, in some areas of Brazil where sand flies do not occur, this disease is found in humans and dogs. There has been speculation that ticks might play a role in transmission of canine visceral leishmaniasis and the DNA of Leishmania spp. has been reported in whole ticks. We investigated the presence of Leishmania spp. promastigotes in the intestines, ovaries, and salivary glands of Rhipicephalus sanguineus ticks collected from tick-infested dogs in two cities of Brazil. We used 66 dogs that tested positive and 33 that tested negative for Leishmania spp. according to direct cytological examination assays. Ten ticks were collected from each dog and dissected to collect the intestines, ovaries, and salivary glands for immunohistochemistry (IHC) and diagnostic real-time polymerase chain reaction (RT-PCR). IHC results showed Leishmania spp. in 98, 14, and 8 % of the intestines, ovaries, and salivary glands, respectively. Real-time PCR showed that 89, 41, and 33 % of the tick intestine, ovary, and salivary glands, respectively, were positive for Leishmania spp. The verification of promastigotes of Leishmania spp. by two independent techniques in ticks collected from these urban region dogs showed that there is need for clarification of the role of ticks in the transmission of canine visceral leishmaniasis in Brazil.

Identification of Leishmania spp. promastigotes in the intestines, ovaries, and salivary glands of Rhipicephalus sanguineus actively infesting dogs

Sand flies are recognized as the major vector of canine visceral leishmaniasis. However, in some areas of Brazil where sand flies do not occur, this disease is found in humans and dogs. There has been speculation that ticks might play a role in transmission of canine visceral leishmaniasis and the DNA of Leishmania spp. has been reported in whole ticks. We investigated the presence of Leishmania spp. promastigotes in the intestines, ovaries, and salivary glands of Rhipicephalus sanguineus ticks collected from tick-infested dogs in two cities of Brazil. We used 66 dogs that tested positive and 33 that tested negative for Leishmania spp. according to direct cytological examination assays. Ten ticks were collected from each dog and dissected to collect the intestines, ovaries, and salivary glands for immunohistochemistry (IHC) and diagnostic real-time polymerase chain reaction (RT-PCR). IHC results showed Leishmania spp. in 98, 14, and 8 % of the intestines, ovaries, and salivary glands, respectively. Real-time PCR showed that 89, 41, and 33 % of the tick intestine, ovary, and salivary glands, respectively, were positive for Leishmania spp. The verification of promastigotes of Leishmania spp. by two independent techniques in ticks collected from these urban region dogs showed that there is need for clarification of the role of ticks in the transmission of canine visceral leishmaniasis in Brazil.

Rhipicephalus (Boophilus) microplus aquaporin as an effective vaccine antigen to protect against cattle tick infestations.

BACKGROUND: Vaccination as a control method against the cattle tick, Rhipicephalus (Boophilus) microplus has been practiced since the introduction of two products in the mid-1990s. There is a need for a vaccine that could provide effective control of R. microplus in a more consistent fashion than existing products. During our transcriptome studies of R. microplus, several gene coding regions were discovered to encode proteins with significant amino acid similarity to aquaporins. METHODS: A cDNA encoding an aquaporin from the cattle tick, Rhipicephalus microplus, was isolated from transcriptomic studies conducted on gut tissues dissected from fully engorged adult female R. microplus. RESULTS: Bioinformatic analysis indicates this aquaporin, designated RmAQP1, shows greatest amino acid similarity to the human aquaporin 7 family. Members of this family of water-conducting channels can also facilitate the transport of glycerol in addition to water. The efficacy of this aquaporin as an antigen against the cattle tick was explored in cattle vaccine trials conducted in Brazil. A cDNA encoding a significant portion of RmAQP1 was expressed as a recombinant protein in Pichia pastoris, purified under native conditions using a polyhistidine C-terminus tag and nickel affinity chromatography, emulsified with Montanide adjuvant, and cattle vaccinated intramuscularly. The recombinant protein provided 75% and 68% efficacy in two cattle pen trials conducted in Campo Grande, Brazil on groups of 6 one year old Holstein calves. CONCLUSION: The effectiveness of this vaccine in reducing the numbers of adult female ticks shows this aquaporin antigen holds promise as an active ingredient in cattle vaccines targeted against infestations of R. microplus.

Preparation of extruded polyethylene/chitosan blends compatibilized with polyethylene-graft-maleic anhydride.

Novel films of polyethylene and chitosan were obtained using extrusion. These polymers have interesting properties, and processing them with methods that are of high use in the industry, such as the extrusion method, can have a significant effect on the potential applications of these materials. The individual materials were thermally characterized; after this, extruded films of low density polyethylene and chitosan mixtures were prepared with the addition of polyethylene-graft-maleic anhydride as a compatibilizer for the blends, and glycerol, as a plasticizer for chitosan. The use of compatibilizer and plasticizer agents improved the processability and compatibility of the mixtures, as well as their mechanical properties, as revealed by mechanical property measurements and scanning electron microscopy. It was possible to prepare blends with a maximum chitosan content of 20 wt%. The material stiffness increased with the increase of chitosan in the sample. FTIR studies revealed the existence of an interaction between the compatibilizer and chitosan.

Restoration of the normal Clara cell phenotype after chronic allergic inflammation.

Bronchiolar Clara cells play a critical role in lung homoeostasis. The main goal of this study was to evaluate the effects of chronic allergy on these cells and the efficacy of budesonide (BUD) and montelukast (MK) in restoring their typical phenotypes after ovalbumin-induced chronic allergy in mice. Chronic allergy induced extensive bronchiolar Alcian blue-periodic acid-Schiff (AB/PAS)-positive metaplasia. In addition, cells accumulated numerous big electron-lucent granules negative for Clara cell main secretory protein (CC16), and consequently, CC16 was significantly reduced in bronchoalveolar lavage. A concomitant reduction in SP-D and CYP2E1 content was observed. The phenotypic changes induced by allergy were pharmacologically reversed by both treatments; MK was more efficient than BUD in doing so. MK decreased AB/PAS reactivity to control levels whereas they remained persistently elevated after BUD. Moreover, most non-ciliated cells recovered their normal morphology after MK, whereas for BUD normal cells coexisted with 'transitional' cells that contained remnant mucous granules and stained strongly for CC16 and SP-D. Glucocorticoids were also less able to reduce inflammatory infiltration and maintained higher percentage of neutrophils, which may have contributed to prolonged mucin expression. These results show that chronic allergy-induced mucous metaplasia of Clara cells affects their defensive mechanisms. However, anti-inflammatory treatments were able to re-establish the normal phenotype of Clara cell, with MK being more efficient at restoring a normal profile than BUD. This study highlights the role of epithelial cells in lung injuries and their contribution to anti-inflammatory therapies.

Protective immunity against tick infestation in cattle vaccinated with recombinant trypsin inhibitor of Rhipicephalus microplus.

The cattle tick, Rhipicephalus microplus, is regarded as the most economically important ectoparasite of livestock globally. Control is achieved primarily through the use of acaricides. This approach is hampered by the development of resistance to commercial acaricides among cattle tick populations. Vaccination against R. microplus infestation is another technology that can be integrated for effective cattle tick control. Proteins belonging to the Kunitz-BPTI family are abundant in cattle tick salivary glands, midgut, and ovaries. These organs are attractive targets for the development of a novel cattle tick vaccine. Efficacy assessment against cattle tick infestation in bovines using a vaccine containing the recombinant form of a member of the Kunitz family from R. microplus produced in a yeast expression system is reported for the first time here. The yeast Pichia pastoris was bioengineered to produce the recombinant version of a trypsin inhibitor that is expressed in cattle tick larvae (rRmLTI). Immunization with rRmLTI afforded 32% efficacy against R. microplus. The estimated molecular weight of rRmLTI was 46 kDa. Structural homology to the native form of the larval trypsin inhibitor was documented by recognition of rRmLTI in Western-blots using polyclonal antibodies from mice immunized with cattle tick larval extract or rRmLTI. Bioinformatics analysis of the partial nucleotide and deduced amino acid sequences indicated that the rRmLTI closely resembles BmTI-6, which is a three-headed Kunitz protein present in cattle tick ovary and fat tissue.

Phenotype changes inherited by crossing pyrethroid susceptible and resistant genotypes from the cattle tick Riphicephalus (Boophilus) microplus.

Dialelic crosses and backcrosses of pyrethroid resistant (RR) and susceptible (SS) Rhipicephalus (Boophilus) microplus tick strains were carried out and the substitution (Phe-Ile) within the sodium channel gene was monitored in order to analyze the effects of the genotype on the pyrethroid resistance phenotype as measured by the larval packet test (LPT). Parental strains: susceptible (SS) and resistant (RR); dialelic crosses: RS (♂RR × â™€SS), and SR (♂SS × â™€RR); and backcrosses: RS × SS, RS × RR, SR × SS and SR × RR were infested on 280 kg calves. Resistance type (monogenic or polygenic) and effective dominance were determined based on the discriminant concentration (DC) for cipermethrine (0.5%), deltamethrine (0.09%) and flumethrine (0.01%). Allele specific PCR (AS-PCR) was used for genotyping, looking at a sodium channel mutation (Phe-Ile substitution). The mortality rates and allele frequency of susceptible and pyrethroid resistant reference strains were 0% mortality and 90% RR alleles for resistant strain, and 100% mortality and 0% RR alleles as measured by the larval packet test (LPT) and allele specific PCR (AS-PCR) respectively. Backcrossed strain SR × RR showed an effective dominance (D(ML)) of 0.605 for cypermethrin, 0.639 for deltamethrin and 0.498 for flumethrin, while survival of backcrosses RS × SS, RS × RR and SR × SS showed a significant tendency to recesivity. Backcrossed strain SR × RR (69.4%) also showed a higher RR genotype frequency with regards to RS × SS (25.5%), RS × RR (36.7%) and SR × SS (32.0%), however, susceptible allele was inherited in general as an incomplete dominant trait. Monogenic inheritance hypothesis was tested and the results showed monogenic inheritance for cypermethrin and flumethrin (P < 0.05) but not for deltamethrin (P > 0.05). However, significant correlation was found between RR genotype and the survival rate for all three pyrethroids used (P < 0.05), suggesting that a single substitution on the sodium channel gene can be responsible for resistance to pyrethroids as a class, due to the high frequency for RR genotypes. Combination with different mutations or metabolic resistance mechanisms cannot be excluded.