Analysing biomarkers in oral fluid from pigs: influence of collection strategy and age of the pig.

BACKGROUND AND OBJECTIVES: Oral fluid (OF) is an easy-to-collect, inexpensive, fast and non-invasive sample to characterize health and welfare status of the pig. However, further standardisation of the collection methods is needed in order to use it regularly in veterinary practice. Cotton ropes are routinely used to collect OF for pathogen detection but they may not be optimal for biomarker analysis due to sample contamination. This study compared two methods (cotton ropes and sponges) to collect porcine OF for biomarker analysis. A panel of 11 biomarkers of stress, inflammation, sepsis, immunity, redox status and general homeostasis was studied. MATERIALS AND METHODS: Eighteen farrow-to-finish pig farms were included in the study. In each farm, three (for sponges) or four pens of pigs (for ropes) were sampled at four age categories: the week after weaning (5 weeks), before (11-12 weeks) and after (12-13 weeks) moving to finisher facility and the week before slaughter (22-25 weeks). In total, 288 OF samples were collected with cotton ropes and 216 with sponges and analysed for the biomarkers: cortisol, alpha-amylase, oxytocin (stress), haptoglobin (inflammation), procalcitonin (sepsis), adenosine deaminase, immunoglobulin G (immune system), ferric reducing antioxidant power (redox status), and creatine kinase, lactate dehydrogenase and total protein (general homeostasis). Samples were also scored visually for dirtiness using a score from 1 (clean) to 5 (very dirty). RESULTS: Rope-collected OF had higher levels of dirtiness (3.7 ± 0.04) compared to sponge-collected OF (2.7 ± 0.15) and had higher values than sponges for cortisol, procalcitonin, oxytocin, haptoglobin, total protein, lactate dehydrogenase and ferric reducing antioxidant power. All biomarkers decreased in value with age. Immunoglobulin G did not perform well for any of the two collection methods. DISCUSSION AND CONCLUSION: The results showed a clear effect of age on the biomarkers in OF collected with both, sponges or ropes. Sponges provided a cleaner sample than cotton ropes for biomarker analysis. Both methods are easy to apply under the commercial conditions in pig farms although sponges may take more time in early weaner stages. From a practical point of view, sampling with sponges achieved the best combination of reduced sampling time and low contamination.

Changes in salivary biomarkers of stress, inflammation, redox status, and muscle damage due to Streptococcus suis infection in pigs.

BACKGROUND: Streptococcus suis (S. suis) is a Gram-positive bacteria that infects pigs causing meningitis, arthritis, pneumonia, or endocarditis. This increases the mortality in pig farms deriving in severe economic losses. The use of saliva as a diagnostic fluid has various advantages compared to blood, especially in pigs. In this study, it was hypothesized that saliva could reflect changes in different biomarkers related to stress, inflammation, redox status, and muscle damage in pigs with S. suis infection and that changes in these biomarkers could be related to the severity of the disease. RESULTS: A total of 56 growing pigs from a farm were selected as infected pigs (n = 28) and healthy pigs (n = 28). Results showed increases in biomarkers related to stress (alpha-amylase and oxytocin), inflammation (haptoglobin, inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4), total protein, S100A8-A9 and S100A12), redox status (advanced oxidation protein producs (AOPP)) and muscle damage (creatine kinase (CK), CK-MB, troponin I, lactate, aspartate aminotransferase, and lactate dehydrogenase). An increase in adenosine deaminase (ADA), procalcitonin, and aldolase in infected animals were also observed, as previously described. The grade of severity of the disease indicated a significant positive correlation with total protein concentrations, aspartate aminotransferase, aldolase, and AOPP. CONCLUSIONS: This report revealed that S. suis infection caused variations in analytes related to stress, inflammation, redox status, and muscle damage in the saliva of pigs and these can be considered potential biomarkers for this disease.

Childhood Leukemia Survival in the US-Mexico Border: Building Sustainable Leukemia Care Using Health Systems Strengthening Models.

PURPOSE: Pediatric leukemia outcomes are poor in most low- and middle-income countries (LMICs) and exacerbated by health care systems ill equipped to manage cancer. Effective leukemia management in LMICs involves curating epidemiologic data; providing health care workforce specialty training; developing evidence-based treatments and supportive care programs; safeguarding access to medications and equipment; providing patient and family psychosocial, financial, and nutritional support; partnering with nongovernmental organizations, and ensuring treatment adherence. METHODS: In 2013, through a partnership between North-American and Mexican institutions, we used the WHO Framework for Action, a health systems strengthening model to implement a leukemia care sustainable program aimed at improving acute lymphoblastic leukemia (ALL) outcomes at a public hospital in Mexico. We prospectively assessed clinical features, risk classification, and survival outcomes in children with ALL at Hospital General-Tijuana from 2008 to 2012 (preimplementation) and from 2013 to 2017 (postimplementation). We also evaluated program sustainability indicators. RESULTS: Our approach led to a fully-staffed leukemia service, sustainable training programs, evidence-based and data-driven projects to improve clinical outcomes, and funding for medications, supplies, and personnel through local partnerships. Preimplementation and postimplementation 5-year overall survival for the entire cohort of children with ALL, children with standard-risk ALL, and children with high-risk ALL improved from 59% to 65% (P = .023), 73% to 100% (P < .001), and 48% to 55% (P = .031), respectively. All sustainability indicators improved between 2013 and 2017. CONCLUSION: Using the health systems strengthening WHO Framework for Action model, we improved leukemia care and survival in a public hospital in Mexico across the US-Mexico border. We provide a model for the development of similar programs in LMICs to sustainably improve leukemia and other cancer outcomes.

Dexamethasone intravitreal implant (Ozurdex) in diabetic macular edema: real-world data versus clinical trials outcomes.

AIM: To investigate the safety and efficacy of intravitreal dexamethasone implants (Ozurdex®/DEX) in patients with diabetic macular edema (DME) either naïve or non-naïve to anti-VEGF therapies who switched to DEX implant independent of response to anti-vascular endothelial growth factors (anti-VEGFs). METHODS: This was an audit retrospective review of medical records of patients with DME who switched to the DEX intravitreal implant. Patients were divided into 2 groups: patients naïve to antiangiogenic therapy and patients who were previously treated with anti-VEGFs. Data regarding demographics, changes in mean best-corrected visual acuity (BCVA), central macular thickness (CMT), and intraocular pressure (IOP) was collected over 6mo. The demographic data mean changes in BCVA, CMT, and IOP were compared. Six-month follow-up data of 47 patients (57 eyes), who either switched to DEX implant irrespective of response to previous treatments or were treatment naïve before receiving DEX implant, was collected. RESULTS: Improvement in mean BCVA was observed from 1-4mo after injection with a decreased effect at month 6 as expected, with better outcomes in naïve compared to non-naïve patients. A statistically relevant decrease in mean CMT was observed during the follow-up period. An increase in mean IOP was observed in the first 2mo after DEX therapy. The mean number of injections of the overall population during the 6mo was 1.3. A subgroup analysis showed no relevant difference between phakic versus pseudophakic patients relative to measured outcomes. There was no cataract progression during the follow-up period and no adverse events reported. CONCLUSION: This real-life setting study shows that intravitreal DEX implant is effective and safe. The timings of greater therapeutic impact are concordant with previous studies and suggest that earlier treatment with corticosteroids may have an additional benefit in naïve patients.

Macular edema secondary to retinal vein occlusion in a real-life setting: a multicenter, nationwide, 3-year follow-up study.

PURPOSE: To evaluate the presence of macular edema secondary to retinal vein occlusion (RVO)-both central retinal vein occlusion (CRVO) and branch retinal vein occlusion (BRVO)-3 years after diagnosis in patients who underwent intravitreal therapy and to identify potential prognostic factors and biomarkers of persistent macular edema. METHODS: National multicenter, observational, exploratory, retrospective cohort study of 104 consecutive patients with macular edema secondary to RVO diagnosed from January 2014 to December 2015 with minimum 3-year follow-up time. Data analyzed included best-corrected visual acuity (BCVA), clinical and demographic data, and spectral domain optical coherence tomography parameters. RESULTS: At final observation, median baseline central retinal thickness significantly improved from baseline 538 to 290 µm (p < 0.001) and complete macular edema resolution was achieved in 51.0% of patients (56.3% and 42.5% in BRVO and CRVO patients, respectively). BCVA also improved (p < 0.01). Logistic regression analysis revealed a relationship between recurrence of macular edema and disorganization of retinal inner layers (DRIL) at baseline (odds ratio = 2.88; p = 0.013). CONCLUSION: Good long-term anatomical and functional outcomes are achieved with intravitreal treatments in RVO patients. Anatomical success and visual gains seen in the first year were maintained throughout the entire follow-up, though DRIL is a major risk factor for recurrence.

Improving Pediatric Neuro-Oncology Survival Disparities in the United States-Mexico Border Region: A Cross-Border Initiative Between San Diego, California, and Tijuana, Mexico.

PURPOSE: Treatment of children with CNS tumors (CNSTs) demands a complex, interdisciplinary approach that is rarely available in low- and middle-income countries. We established the Cross-Border Neuro-Oncology Program (CBNP) between Rady Children's Hospital, San Diego (RCHSD), and Hospital General, Tijuana (HGT), Mexico, to provide access to neuro-oncology care, including neurosurgic services, for children with CNSTs diagnosed at HGT. Our purpose was to assess the feasibility of the CBNP across the United States-Mexico border and improve survival for children with CNSTs at HGT by implementing the CBNP. PATIENTS AND METHODS: We prospectively assessed clinicopathologic profiles, the extent of resection, progression-free survival, and overall survival (OS) in children with CNSTs at HGT from 2010 to 2017. RESULTS: Sixty patients with CNSTs participated in the CBNP during the study period. The most common diagnoses were low-grade glioma (24.5%) and medulloblastoma (22.4%). Of patients who were eligible for surgery, 49 underwent resection at RCHSD and returned to HGT for collaborative management. Gross total resection was achieved in 78% of cases at RCHSD compared with 0% at HGT (P < .001) and was a predictor of 5-year OS (hazard ratio, 0.250; 95% CI, 0.067 to 0.934; P = .024). Five-year OS improved from 0% before 2010 to 52% in 2017. CONCLUSION: The CBNP facilitated access to complex neuro-oncology care for underserved children in Mexico through binational exchanges of resources and expertise. Survival for patients in the CBNP dramatically improved. Gross total resection at RCHSD was associated with higher OS, highlighting the critical role of experienced neurosurgeons in the treatment of CNSTs. The CBNP model offers an attractive alternative for children with CNSTs in low- and middle-income countries who require complex neuro-oncology care, particularly those in close proximity to institutions in high-income countries with extensive neuro-oncology expertise.

Ranibizumab for the Treatment of Diabetic Macular Oedema in the Real-World Clinical Setting in Portugal: A Multicentre Study.

PURPOSE: The purpose of this study was to evaluate the 2-year outcome of ranibizumab for diabetic macular oedema (DME) in the real-life clinical practice of five ophthalmology departments of the National Health Service (NHS) in Portugal. METHODS: This is a retrospective multicentre study. The clinical records on consecutive patients with DME from clinical practice treated with 0.5 mg intravitreal ranibizumab and followed up for 24 months were reviewed. Efficacy outcomes comprised the change in best corrected visual acuity (BCVA) and central macular thickness (CMT) evaluated by SD-OCT. Multivariate regression analysis was performed to explore predictors of BCVA. RESULTS: A total of 122 eyes of 93 patients were included. The median BCVA change by 24 months was +5.0 letters (IQR 12.0) (p < 0.001) and the CMT change was -89.0 µm (IQR 165.0) (p < 0.001). By 24 months, 21.4% of the eyes had gained ≥15 letters and 8.6% had lost ≥15 letters. The median number of injections given during follow-up was 5.0 (IQR 4.0). A greater baseline CMT and a more disrupted status of the external limiting membrane were predictive of worse BCVA at 24 months (p ≤ 0.015). CONCLUSION: DME treatment with ranibizumab in the Portuguese NHS is associated with anatomic and functional improvement by 2 years; however, our results are below those reported in major clinical trials, and undertreatment is probably the cause.

Addressing regional disparities in pediatric oncology: Results of a collaborative initiative across the Mexican-North American border.

BACKGROUND: Cancer is emerging as a major cause of childhood mortality in low- and middle-income countries. In Mexico, cancer is the number one cause of death in children aged 5-14. Until recently, many children with cancer from Baja California, Mexico, went untreated. We reasoned that an initiative inspired by the St. Jude Children's Research Hospital (SJCRH) "twinning" model could successfully be applied to the San Diego-Tijuana border region. In 2008, a twinning project was initiated by Rady Children's Hospital, SJCRH, and the General Hospital Tijuana (GHT). Our aim was to establish a pediatric oncology unit in a culturally sensitive manner, adapted to the local healthcare system. PROCEDURE: An initial assessment revealed that despite existence of basic hospital infrastructure at the GHT, the essential elements of a pediatric cancer unit were lacking, including dedicated space, trained staff, and uniform treatment. A 5-year action plan was designed to offer training, support the staff financially, and improve the infrastructure. RESULTS: After 7 years, accomplishments include the opening of a new inpatient unit with updated technology, fully trained staff, and a dedicated, interdisciplinary team. Over 700 children have benefited from accurate diagnosis and treatment. CONCLUSIONS: Initiatives that implement long-term partnerships between institutions along the Mexican-North American border can be highly effective in establishing successful pediatric cancer control programs. The geographic proximity facilitated accelerated training and close monitoring of project development. Similar initiatives across other disciplines may benefit additional patients and synergize with pediatric oncology programs to reduce health disparities in underserved areas.

Diagnostic performance of fusion of myocardial perfusion imaging (MPI) and computed tomography coronary angiography.

BACKGROUND: We evaluated the incremental diagnostic value of fusion images of coronary computed tomography angiography (CTA) and myocardial perfusion imaging (MPI) over MPI alone or MPI and CTA side-by-side to identify obstructive coronary artery disease (CAD > 50% stenosis) using invasive coronary angiography (ICA) as the gold standard. METHODS: 50 subjects (36 men; 56 +/- 11 years old) underwent rest-stress MPI and CTA within 12-26 days of each other. CTAs were performed with multi-detector CT-scanners (31 on 64-slice; and 19 on 16-slice). 37 patients underwent ICA while 13 subjects did not because of low (<5%) pre-test likelihood (LLK) of disease. Three blinded readers scored the images in sequential sessions using (1) MPI alone (2) MPI and CTA side-by-side, (3) fused CTA/MPI images. RESULTS: One or more critical stenoses during ICA were found in 28 patients and non-critical stenoses were found in 9 patients. MPI, side-by-side MPI-CTA, and fused CTA/MPI showed the same normalcy rate (NR:13/13) in LLK subjects. The fusion technique performed better than MPI and MPI and CTA side-by-side for the presence of CAD in any vessel (overall area under the curve (AUC) for fused images: 0.89; P = .005 vs MPI, P = .04 vs side-by-side MPI-CTA) and for localization of CAD to the left anterior descending coronary artery (AUC: 0.82, P < .001 vs MPI; P = .007 vs side-by-side MPI-CTA). There was a non-significant trend for better detection of multi-vessel disease with fusion. CONCLUSIONS: Using ICA as the gold standard, fusion imaging provided incremental diagnostic information compared to MPI alone or side-by-side MPI-CTA for the diagnosis of obstructive CAD and for localization of CAD to the left anterior descending coronary artery.

Prognostic performance of quantitative PET tools for stratification of patients with ischemic cardiomyopathy undergoing myocardial viability assessment.

OBJECTIVES: This study was performed to determine the prognostic performance of quantitative PET tools in the stratification of patients with ischemic cardiomyopathy undergoing myocardial viability assessment. METHODS: We applied four different quantitative tools to 104 consecutive patients with coronary artery disease and previous myocardial infarction who had undergone rest Rb/gated F-fluorodeoxyglucose (FDG) PET, to assess myocardial viability for potential revascularization. One of these tools was based on the FDG study alone and the other three tools assessed the extent of match/mismatch defects using FDG in comparison with a perfusion reference database. The four quantitative tools used in this research to define viability were (i) FDG alone, which calculates the percentage of left ventricular myocardium (LVM) that is above the 50% of the maximum LVM FDG counts, (ii) low flow match/mismatch, which determines the area with a 5% increase in normalized FDG counts in relation to defined resting perfusion defects as compared with a reference database, (iii) all regions match/mismatch, which computes the area with a 10% increase in normalized FDG counts in relation to the left ventricle resting perfusion distribution, and (iv) percentage max FDG match/mismatch, which defines the area with FDG uptake greater than 60% of the maximum LVM FDG counts within defined perfusion defects as determined by the reference database. The primary endpoint for this analysis was cardiac death. RESULTS: During the follow-up period (22+/-14 months), 19 patients (18%) died; in 17 of these the cause of death was cardiac. Using univariate analysis, none of the methods were predictive of cardiac death. Receiver operating characteristic analysis defined the optimal thresholds for the extent of myocardial viability for the four tools in the prediction of cardiac death: FDG alone=20%, low flow match/mismatch=15%, all regions match/mismatch=35%, and percentage max FDG match/mismatch=20%. A censored survival analysis using a Kaplan-Meier method showed a statistically significant difference between patients with cardiac death and those with no cardiac death using only the low flow match/mismatch (hazard ratio=0.29, P=0.01) and percentage max FDG match/mismatch criteria (hazard ratio=0.23, P=0.005) tools. CONCLUSION: The low flow match/mismatch and percentage max FDG match/mismatch quantitative PET tools are useful for prognostic stratification of patients with ischemic cardiomyopathy undergoing myocardial viability assessment.

Normal values and prospective validation of transient ischaemic dilation index in 82Rb PET myocardial perfusion imaging.

BACKGROUND: The use of Rb positron emission tomography (PET) for the diagnosis of coronary artery disease (CAD) has increased in recent years but the role of some of the traditional parameters used in SPECT for the diagnosis of CAD, such as transient ischaemic dilation index (TID) of the left ventricle, have not been validated in PET studies. METHODS AND RESULTS: We studied 95 patients who had undergone rest/pharmacological stress Rb PET scans. Thirty of these patients (18 female and 12 male) who had less than 5% likelihood of CAD (LLK) based on sequential Bayesian analysis, were used to determine the normal limits of TID index in this protocol. The remaining 65 patients (33 female and 32 male) underwent coronary angiography within 15 days of the cardiac PET scan. This second group of patients was used to validate the TID normal limits determined in the first group. In LLK patients mean TID index was 1.01+/-0.07 and there were no significant differences between genders. The TID index upper normal limit was 1.15 and was calculated as mean+2 SD. Using this cut-off point, TID index had high specificity and PPV in the diagnosis of single vessel CAD (100% and 100% respectively) and multiple vessel CAD (93% and 85%, respectively). CONCLUSION: Our results indicate that elevated TID index is a specific, although not sensitive marker of single and multiple vessel CAD in pharmacologically stressed Rb PET myocardial perfusion studies.

Metabolic syndrome and ischemic heart disease in gout.

BACKGROUND: For decades, gout has been associated with several metabolic abnormalities and with ischemic heart disease (IHD). OBJECTIVE: Our aim was to determine the prevalence of metabolic syndrome by Adult Treatment Panel III criteria (ATP III) and ischemic heart disease (IHD) by electrocardiogram (EKG) and/or single photon emission computed tomography (SPECT) in patients with gout. METHODS: We included 64 consecutive outpatients with primary gout, but no history of IHD, attending our clinic for the first time. Demographic and clinical data were recorded and resting electrocardiogram, lipid profile, fasting insulin, and SPECT with Tc sestamibi were performed. Metabolic syndrome was defined according to ATP III criteria (> or =3 of the following data: 1) hyperglycemia (fasting glucose > or =110 mg/dL) or previous diagnosis of diabetes mellitus, 2) hypertension (> or =130/85 mm Hg) or previous diagnosis, 3) high-density lipoprotein (HDL) <40 mg/dL (men) or <50 mg/dL (women), 4) triglycerides > or =150 mg/dL, and 5) obesity. RESULTS: IHD was diagnosed in 10 patients (16%); 2 had EKG changes compatible with previous silent myocardial necrosis and the other 8 had abnormal SPECT. The prevalence of metabolic syndrome was 82%, all patients had at least 1 metabolic abnormality, but all the patients with IHD had metabolic syndrome (3 criteria according with ATP III). Patients with IHD differed from those without IHD in the percentage of HDL levels <40 mg/dL (100% vs. 82%; P = 0.05) as well as glucose and insulin levels in the fasting state (129.3 +/- 6.1 mg/dL vs. 92.7 +/- 16.7 mg/dL; P = 0.000; and 21.1 +/- 6.0 vs. 17.5 +/- 8.6 UI/mL; P = 0.03) and low-density lipoproteins (143.9 +/- 21.3 mg/dL vs. 118.2 +/- 47.7 mg/dL; P = 0.014). In contrast, serum creatinine and urea (1.02 +/- 0.13 mg/dL vs. 1.5 +/- 1.5 mg/dL; P = 0.024; and 33.9 +/- 9.3 mg/dL vs. 48.7 +/- 46.1 mg/dL; P = 0.039) and creatinine clearance <50 mL/min (10% vs. 37%; P = 0.06) were higher in patients without IHD. CONCLUSIONS: In this work, metabolic syndrome was very common among patients with gout. Sixteen percent of the patients, although previously asymptomatic, had IHD, they all had metabolic syndrome. Gouty patients frequently first seek medical care from a rheumatologist. The rheumatologist can have an important role in detecting metabolic syndrome and risk factors for cardiovascular disease.