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Abstract Background Porcine circovirus type 2 (PCV2) has been associated with several disease complexes, including reproductive failure. The aim of this study was to identify the subtypes of PCV2 that are associated with reproductive failure in pigs from the State of São Paulo, Brazil and to investigate co-infections with other infectious organisms. Findings Samples of 168 aborted foetuses or mummified foetuses from five farrow-to-finish swine farms known to be infected with PCV2 and located in the State of São Paulo were tested for PCV2 by polymerase chain reaction (PCR). Positive samples were additionally tested for porcine parvovirus (PPV), Leptospira spp. and Brucella spp. by PCR. PCV2 was detected in 18 of the samples (10.7%). PPV, Brucella spp. and Leptospira spp were found in 2, 10 and 0 cases, respectively. Eleven PCV2 strains were sequenced and determined to be either genotype 2a (n = 1) or 2b (n = 10). Conclusions The findings indicate that the frequency of PCV2 infections in aborted porcine foetuses from the State of São Paulo is rather low (10.7%) and that co-infection with other pathogens is common and may be involved in PCV2 associated reproductive failure. No repeatable, characteristic amino acid motifs for regions of the PCV2 capsid protein seemed to be associated with abortion in sows.

Aiming to study the performance, carcass characteristics, nutrient digestibility, blood parameters, salivary cortisol levels, and economic viability of pigs administered aspartic protease, a total of 135 pigs were housed in pens in a randomized block design, divided into five treatments with nine replications. The experimental diets were positive control (PC), basic diet with a 5.0% reduction in protein and amino acid requirements; negative control (NC) with a 7.5% reduction in protein and amino acid requirements; NC + 100 g/mT of aspartic protease (NC100); NC + 150 g/mT of aspartic protease (NC150); and NC + 200 g/mT of neutral serine protease (NC200). The inclusion of protease, independently of the source and amount, increased the average daily weight gain (P < 0.05) of animals compared with the control treatments (PC and NC), improved feed conversion (P < 0.05) in early stages, and improved diet digestibility (P < 0.05) compared with the PC. Treatment with NC150 and NC200 resulted in greater carcass weights (P < 0.05) than treatment with the PC. NC100 led to a greater carcass yield than PC (P < 0.05), and NC150 resulted in a greater loin eye area than PC (P < 0.05). No differences (P > 0.05) in the blood parameters or salivary cortisol levels were found. Regarding economic viability, proteases increased the profitability, with NC150 leading to the best results. Thus, the use of aspartic proteases is recommended to improve performance and further facilitate pork production., (© 2024. The Author(s).)

Liquid-liquid phase separation is a phenomenon within biology whereby proteins can separate into dense and more dilute phases with distinct properties. Three antibodies that undergo liquid-liquid phase separation were characterized in the protein-rich and protein-poor phases. In comparison to the protein-poor phase, the protein-rich phase demonstrates more blue-shift tryptophan emissions and red-shifted amide I absorbances. Large changes involving conformational isomerization around disulfide bonds were observed using Raman spectroscopy. Amide I and protein fluorescence differences between the phases persisted to temperatures above the critical temperature but ceased at the temperature at which aggregation occurred. In addition, large changes occurred in the structural organization of water molecules within the protein-rich phase for all three antibodies. It is hypothesized that as the proteins have the same chemical potential in both phases, the protein viscosity is higher in the protein-rich phase resulting in slowed diffusion dependent protein aggregation in this phase. For all three antibodies we performed accelerated stability studies and found that the protein-rich phase aggregated at the same rate or slower than the protein-poor phase., Competing Interests: Declaration of Interests RE and CKK are employees of AstraZeneca and may own stock or stock options. The remaining authors have no interests to disclose related to this work., (Copyright © 2022 American Pharmacists Association. Published by Elsevier Inc. All rights reserved.)

The objective of this work was to evaluate the effect of a phytogenic compound for pigs in the growing and finishing phases as a possible substitute for ractopamine. A total of 140 pigs with an average initial weight of 48.8 kg ± 5.9 kg were used, distributed in a randomized block design, in a 3 × 2 factorial scheme (control diet (CONT), diet with inclusion of 2.5 kg per ton of a phytogenic compound (PC), and diet with 10 ppm of ractopamine (RAC), and two sexes: gilts and barrows), distributed in eight pens per treatment. The performance parameters were measured, and, at the end of the experimental period, the animals were slaughtered for carcass characteristics and pork quality analysis. The animals consuming RAC showed a better feed conversion, 4% improvement in relation to the group with the PC (p < 0.05). For daily weight gain, the animals supplemented with the PC showed 4.46% lower gain compared to RAC, and 3% greater gain compared to the CONT (p < 0.05). The animals that consumed the PC showed 5.6% lower shear force of the pork (p < 0.05) in relation to the CONT group and 29% lower in relation to the RAC group. The TBARS value presented a significant difference (p < 0.05), the CONTT group was 29% higher than the RAC, and the PC was 15.5% higher than the RAC. For chroma, the pork of the RAC group was 14% lower than the CONTT group and 10.3% less than the PC. There was no significant difference for the carcass parameters. It was concluded that the pigs in the ractopamine group presented the best performance; however, the phytogenic compound can be used against ractopamine’s restriction because it improves daily weight gain and promotes a softer and less pale meat when compared with ractopamine., Competing Interests: The authors declare no conflict of interest.

Objectives: To compare different approaches for the expression of an anti-PCSK9 biosimilar monoclonal antibody (mAb) in CHO cells using IRES-mediated tricistronic plasmid vectors combining different signal peptides, IRES elements and selection markers., Results: Transient transfection indicated a similar level of secreted mAb 48 h post-transfection for all constructs. However, transfections carried out with circular plasmids showed a higher expression than with linearized plasmids. After two months under selection pressure, only part of the transfected pools recovered. The cultures co-transfected using two antibiotics as selection markers for double selection did not recover. Growth, metabolism and mAb production profiles of the only part of the transfected pools recovered resulting stable pools were compared and the stable pool transfected with circular L1-LC-IRES-H7-HC-IRES-NEO plasmid was chosen for further studies, due to higher cell growth and mAb production. Critical quality attributes of the protein A-purified mAb such as purity, homogeneity, binding affinity to PCSK9, and amino acid sequence were assessed confirming the success of the approach adopted in this study., Conclusions: The expression platform proposed showed to be efficient to produce a high-quality anti-PCSK9 mAb in stable CHO cell pools and provides benchmarks for fast production of different mAbs for characterization, formulation studies and pre-clinical investigation.

In 2016, the Zambian government made cancer control a national priority and released a National Cancer Control Strategic Plan for 2016 to 2021, which focuses on malignancies of the breast, cervix, and prostate, and retinoblastoma. The plan calls for a collective reduction in the cancer burden by 50%. In support of this vision, Susan G. Komen sponsored a consultative meeting in Lusaka, Zambia, in September 2017 to bring together the country's main breast cancer stakeholders and identify opportunities to improve breast cancer control. The recommendations generated during the discussions are presented. There was general agreement that the first step toward breast cancer mortality reduction should consist of implementation of early detection service platforms focused on women who are symptomatic. Participants also agreed that the management of all components of the national breast cancer control program should be integrated and led by the Ministry of Health. As much as possible, early detection and treatment services presently offered by the Cervical Cancer Prevention Program of Zambia and Cancer Diseases Hospital should be leveraged. Efforts are under way through multiple stakeholders to implement the following recommendations: development of national guidelines for the early diagnosis of breast cancer, training of breast surgeons, implementation of early detection and surgical treatment service platforms at the district-hospital level, and epidemiologic research, including the improvement of electronic recording mechanisms.

Metastatic breast cancer (mBC) is an area with high unmet need across the world. Despite significant progress to meet patient need, current breast cancer (BC) policies fail to recognize the unique challenges of patients who are further along the cancer patient journey. This analysis aims to understand BC/mBC policy development at a global level and identify opportunities for further development. A comprehensive analysis of National Cancer Control Plans (NCCPs) policies and programs was conducted across 16 countries, which represent a diverse range of healthcare systems, economies and geographic regions. Examples of promising practices, implemented or initiated by civil society, are provided to demonstrate successful methods to address the identified policy gaps. The analysis finds that disparities in BC policy development exist across and within countries. Progress in BC policy is fragmented and skewed towards the early part of the patient journey e.g. awareness and stakeholder education, with key gaps remaining in diagnosis and patient identification. In addition, access to innovative mBC treatments, ongoing support and palliative care remain a challenge, while care coordination is limited due to inefficient referrals. Although government and policymaker action is fundamental, collaboration between different stakeholders is imperative to address unmet needs of BC/mBC patients alike. Policy initiatives and promising practices that demonstrate successful multi-stakeholder engagement can be replicated or used to inform further advocacy and policy development with the aim to address patient unmet needs., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)

High-throughput screening (HTS) systems have emerged as important tools to provide fast and low cost evaluation of several conditions at once since it requires small quantities of material and sample volumes. These characteristics are extremely valuable for experiments with large number of variables enabling the application of design of experiments (DoE) strategies or simple experimental planning approaches. Once, the capacity of HTS systems to mimic chromatographic purification steps was established, several studies were performed successfully including scale down purification. Here, we propose a method for studying different purification conditions that can be used for any recombinant protein, including complex and glycosylated proteins, using low binding filter microplates.

The pressures to efficiently produce complex biopharmaceuticals at reduced costs are driving the development of novel techniques, such as in downstream processing with straight-through processing (STP). This method involves directly and sequentially purifying a particular target with minimal holding steps. This work developed and compared six different 3-step STP strategies, combining membrane adsorbers, monoliths, and resins, to purify a large, complex, and labile glycoprotein from Chinese hamster ovary cell culture supernatant. The best performing pathway was cation exchange chromatography to hydrophobic interaction chromatography to affinity chromatography with an overall product recovery of up to 88% across the process and significant clearance of DNA and protein impurities. This work establishes a platform and considerations for the development of STP of biopharmaceutical products and highlights its suitability for integration with single-use technologies and continuous production methods. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:931-940, 2017., (© 2017 American Institute of Chemical Engineers.)

Introduction: Proteinuria after kidney transplantation (Tx) has variable incidence and is associated with cardiovascular risk and graft survival., Objective: To evaluate the prevalence of proteinuria after kidney Tx and its associated factors., Methods: The prevalence of PTN was evaluated according to definition ≥ 500 mg/24 hours. Patients were divided into 3 groups: group A, < 500 mg, B, 500-1000 mg and C, > 1000 mg. We tested the association between PTN and: age/gender of the donor and recipient, type of donor, delayed graft function, acute rejection, hypertension and creatinine. The variables with a p value < 0.20 in the bivariate analysis were included in a multivariate logistic regression analysis., Results: 173 recipients were evaluated, mean age 39 years, 57.2% male and 60.7% deceased donor. The prevalence of PTN after kidney Tx was 24.3%. The distribution of patients according to PTN was 75.7% for group A, 15.6% for group B and 8.7% for group C. The following factors were associated with higher risk of PTN: male recipients, living donor and hypertension. Creatinine at month 12 moths post-Tx was higher among patients with proteinuria. 60% of patients with PTN ≥ 500 mg/24 hours were treated with ACEI/ARB., Conclusion: The prevalence of PTN after kidney Tx varied between 24.3%, according to the definition used. The male gender of the recipient, living donor and hypertension were associated with the occurrence of PTN after kidney Tx. Blockade of the renin-angiotensin system must be prescribed to more patients.

The present study investigated the predictors of an increased number of visits from individuals with some of the diagnoses noted in chapter F14 of ICD-10, from calls to the emergency psychiatric unit of a general hospital in São Paulo state, Brazil, in the period 2011-2012. Poisson regression models were carried out for the outcome variable, accounting for number of subsequent visits to the psychiatric emergency unit. For the analysis of this outcome we took into account the exposure time of each individual in the study. Our findings point to a population at risk for frequent psychiatric emergency service visits: individuals over 25 years. This population should be targeted for interventions on entry into public healthcare due to increased psychiatric morbidity and greater clinical morbidity already confirmed by previous studies. We discussed the need of these individuals for special attention during the clinical or psychiatric emergency consultation which, unfortunately, may be the access point for the public health system. None of the other variables were related to the outcome of interest, such as those related to the level of individual entry into the care network before and after treatment, and other variables related to medical acts during the visit.

The intracellular parasite Toxoplasma gondii can penetrate any warm-blooded animal cell. Conserved molecular assemblies of host cell plasma membranes should be involved in the parasite-host cell recognition. Lipid rafts are well-conserved membrane microdomains that contain high concentrations of cholesterol, sphingolipids, glycosylphosphatidylinositol, GPI-anchored proteins, and dually acylated proteins such as members of the Src family of tyrosine kinases. Disturbing lipid rafts of mouse peritoneal macrophages and epithelial cells of the lineage LLC-MK2 with methyl-beta cyclodextrin (M β CD) and filipin, which interfere with cholesterol or lidocaine, significantly inhibited internalization of T. gondii in both cell types, although adhesion remained unaffected in macrophages and decreased only in LLC-MK2 cells. Scanning and transmission electron microscopy confirmed these observations. Results are discussed in terms of the original role of macrophages as professional phagocytes versus the LLC-MK2 cell lineage originated from kidney epithelial cells.

The body's adaptive reaction to a stressful event, an allostatic response, involves vigorous physiological engagement with and efficient recovery from stress. Our aim was to investigate the influence of individual predispositions on cardiac responses to and recovery from a standardized psychosocial stress task (Trier Social Stress Task) in peacekeepers. We hypothesized that those individuals with higher trait resilience and those with higher resting vagal control would be more likely to present an allostatic response: a vigorous cardiac response to stress (i.e., reduction in interbeat intervals and heart rate variability (HRV)) coupled with a significant cardiac recovery in the aftermath. Fifty male military personnel with a mean age of 25.4 years (SD ± 5.99) were evaluated after returning from a peacekeeping mission. Electrocardiogram recordings were made throughout the experimental session, which consisted five conditions: basal, speech preparation, speech delivery, arithmetic task, and recovery. Mean interbeat intervals and HRV were calculated for each condition. An Ego-Resilience Scale and resting vagal control assessed individual predispositions. Stress tasks reduced interbeat intervals (tachycardia) and HRV in comparison with basal, with return to basal in the aftermath (p < 0.001, for all comparisons). Resilience and resting vagal control correlated positively with cardiac parameters for both stress reactivity and recovery (r ≥ 0.29; p < 0.05). In conclusion, peacekeepers showing higher trait resilience and those with higher resting vagal control presented a more adaptive allostatic reaction characterized by vigorous cardiac response to stress (i.e., tachycardia and vagal withdrawal) and efficient cardiac recovery after stress cessation.

Mycoplasmal pneumonia of swine causes considerable economic losses in porciculture. Diverse agents, such as environmental stress and infectious microorganisms, increase the possibility of infection, and the damage to pulmonary tissue when the infection is established. It is known that Mycoplasma hyopneumoniae has an important role in this disease, in addition to secondary microbial agents, such as Pasteurella multocida and Actinobacillus pleuropneumoniae. However, the characteristics of this disease in farms are well known. In this work we review the interactions among the different microorganisms involved and the immunological strategies utilized to control this disease. The interaction between Mycoplasma hyopneumoniae and P. multocida in experimental pneumonia was reported by us in conventional pigs. M. hyopneumoniae causes mild pneumonia, whereas P. multocida aggravates the pneumonia initiated by M. hyopneumoniae. The disease has been reproduced to test the efficacy of two immunogens, and can also be used to evaluate some antibiotics. A M. hyopneumoniae bacterine administered intraperitoneally conferred more protection than when it was used with adjuvant, although protection was not complete and colonization by M. hyopneumoniae was not prevented, as is claimed to have been the case with other vaccines.

Corneal phlyctenules and catarrhal infiltrates developed in rabbits immunized with Staphylococcus aureus cell walls (CWs) mixed with complete Freund's adjuvant (CFA) after topical challenge with viable S aureus. The immune responses of these rabbits to CWs and ribitol teichoic acid (RTA) before and after immunization with CW/CFA were studied and the results of skin tests with lymphocyte blastogenesis and hemagglutination tests correlated. After immunization of rabbits with CW/CFA, their lymphocytes showed a greater proliferative response to CWs in the lymphocyte blastogenesis test that correlated with delayed hypersensitivity reactions found in skin tests. Both tests suggested that the lymphocytes of immunized rabbits showed cellular immunity to CWs. Skin tests to RTA suggested an Arthus but not a delayed hypersensitivity reaction to this antigen. The results of skin tests and hemagglutination tests suggested that immunized rabbits expressed humoral and immunity to RTA, the major antigenic determinant of S aureus.

This study aimed at identifying and characterizing the alimentary and nutritional situation of different socio-economic strata of the urban population of Ponte Nova, in Minas Gerais State, Brazil. The sample comprised 161 families stratified according to their earnings in terms of the official minimum wage (salário mínimo--SM), classified in 6 stratum. In addition to socio-economic data, the analysis also takes into consideration the nutritional and caloric adequacy of the ingredients of the basic diet as well as the nutritional state of children up to 6 years old, employing the criteria of Gomez and Seoane-Lathan. The results show that 40% of the sample population earn less than 2 SM, while 12% reach and income level above 10 SM. The population considered has only and indirect relation to production and is employed in the services sector. Access to sanitary services is greater for higher income strata, which also feature better living conditions and health resources. The analysis of the basic diet shows that nutritional adequacy increases with increasing income, reaching more adequate levels only in the highest stratum. The level of perception as to what is "sufficient" in terms of diet supports the results of diet adequacy for each stratum. In so far as the nutritional condition of children is concerned it was observed that the prevalence of malnutrition is consistent with socio-economic condition, decreasing within creased earnings and disappearing altogether above the 10 SM range.