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Biologics have improved the treatment of rheumatic diseases, resulting in better outcomes. However, their high cost limits access for many patients in both North America and Latin America. Following patent expiration for biologicals, the availability of biosimilars, which typically are less expensive due to lower development costs, provides additional treatment options for patients with rheumatic diseases. The availability of biosimilars in North American and Latin American countries is evolving, with differing regulations and clinical indications. The objective of the study was to present the consensus statement on biosimilars in rheumatology developed by Pan American League of Associations for Rheumatology (PANLAR). Using a modified Delphi process approach, the following topics were addressed: regulation, efficacy and safety, extrapolation of indications, interchangeability, automatic substitution, pharmacovigilance, risk management, naming, traceability, registries, economic aspects, and biomimics. Consensus was achieved when there was agreement among 80% or more of the panel members. Three Delphi rounds were conducted to reach consensus. Questionnaires were sent electronically to panel members and comments about each question were solicited. Eight recommendations were formulated regarding regulation, pharmacovigilance, risk management, naming, traceability, registries, economic aspects, and biomimics. The recommendations highlighted that, after receiving regulatory approval, pharmacovigilance is a fundamental strategy to ensure safety of all medications. Registries should be employed to monitor use of biosimilars and to identify potential adverse effects. The price of biosimilars should be significantly lower than that of reference products to enhance patient access. Biomimics are not biosimilars and, if they are to be marketed, they must first be evaluated and approved according to established regulatory pathways for novel biopharmaceuticals. • Biologics have improved the treatment of rheumatic diseases. • Their high cost limits access for many patients in both North America and Latin America. • Biosimilars typically are less expensive, providing additional treatment options for patients with rheumatic diseases. • PANLAR presents its consensus on biosimilars in rheumatology

This evidence- and consensus-based guideline on the treatment of psoriasis vulgaris was developed following the EuroGuiDerm Guideline and Consensus Statement Development Manual. The second part of the guideline provides guidance for specific clinical and comorbid situations such as treating psoriasis vulgaris patient with concomitant psoriatic arthritis, concomitant inflammatory bowel disease, a history of malignancies or a history of depression or suicidal ideation. It further holds recommendations for concomitant diabetes, viral hepatitis, disease affecting the heart or the kidneys as well as concomitant neurological disease. Advice on how to screen for tuberculosis and recommendations on how to manage patients with a positive tuberculosis test result are given. It further covers treatment for pregnant women or patients with a wish for a child in the near future. Information on vaccination, immunogenicity and systemic treatment during the COVID-19 pandemic is also provided.

Contains fulltext : 229416.pdf (Publisher’s version ) (Open Access) This evidence- and consensus-based guideline on the treatment of psoriasis vulgaris was developed following the EuroGuiDerm Guideline and Consensus Statement Development Manual. The first part of the guideline includes general information on the scope and purpose, health questions covered, target users and strength/limitations of the guideline. Suggestions for disease severity grading and treatment goals are provided. It presents the general treatment recommendations as well as detailed management and monitoring recommendations for the individual drugs. The treatment options discussed in this guideline are as follows: acitretin, ciclosporin, fumarates, methotrexate, adalimumab, apremilast, brodalumab, certolizumab pegol, etanercept, guselkumab, infliximab, ixekizumab, risankizumab, secukinumab, tildrakizumab and ustekinumab.

Summary This study was performed in Ross 308 chickens aged 1–21 days and aimed to evaluate whether the addition of 25-hydroxycholecalciferol (25(OH)D3) to broiler chicken diets affects their growth performance and immunity. A completely random 2 × 2 factorial arrangement was used with two levels of vitamin D3 and the absence or presence of 25(OH)D3, corresponding to four treatments based on sorghum + soya bean diets: (i) 200 IU of vitamin D3/kg of feed (Diet 1) (NRC, 1994), (ii) Diet 1 + 69 μg of 25(OH)D3/kg of feed (Diet 2), (iii) 5,000 IU of vitamin D3/kg of feed (Diet 3) and (iv) Diet 3 + 69 μg of 25(OH)D3/kg of feed (Diet 4). Each treatment was conducted with six replicates of 10 chickens each. Water and feed was supplied ad libitum. The results showed significantly increased growth and tibia ash (p

The objective of the study was to compare the effects of caudal epidural bupivacaine and dexmedetomidine (DEX) combination, with bupivacaine or DEX plain for perineal analgesia in mares. Six healthy saddle mares weighing 330–370 kg and aged 10–15 years were used in this study. Each mare was assigned to receive three treatments: 0.04 mg/kg 0.25% bupivacaine (BP), 2 μg/kg DEX (DX), or 0.02 mg/kg bupivacaine and 1 μg/kg DEX (BPDX). The order of treatments was randomized. All drugs were injected into the caudal epidural space (Co1-Co2) through a 16-G Tuohy epidural needle. After the epidural injections, heart rate, respiratory rate, arterial blood pressures (systolic, diastolic, and mean), and rectal temperature were measured at 5, 10, 15, 30, 60, 90, and 120 minutes, and after this time, every 60 minutes until the end of the experiments. A subjective score system was used to assess analgesia, behavioral and motor blockade at the same time points. The BPDX treatment produced analgesic action with twice the duration (200 minutes) of the BP treatment (97 minutes), but with an analgesic duration shorter than the DX treatment (240 minutes) in the regions of the tail, perineum, and upper hind limbs in mares. All treatments showed mild motor blockade. No behavioral changes were observed in any of the animals. There was hemodynamic stability without significant changes in respiratory rate for all treatments. Epidural analgesia using DEX alone or the combination of DEX and bupivacaine may be an option for painful obstetric and gynecological procedures in mares.

Background:Follow-up cohorts (observational studies) were initiated consecutively or simultaneously to the development of randomised controlled trials (RCTs) in JIA patients(1,2). They help to identify many complications observed only in clinical practice related to off label use, coadministration of treatments, drug misuse, and occurrence of rare or unexpected event. In addition, observational studies include a higher number of patients with a longer duration of follow-up compared to randomised trials. Hence, they have a higher power to capture the occurrence of serious adverse events (SAE) in daily clinical practice3.Objectives:To estimate the incidence of serious adverse events (SAEs) including serious infections, malignancies, and death in patients with juvenile idiopathic arthritis (JIA) treated with biological agents (BAs) in daily clinical practice, using meta-analysis techniques.Methods:We systematically searched, up to May 2019, Medline and Embase databases for observational studies performed in JIA disease under BAs treatment. Outcomes were SAEs, serious infections, malignancies and all-cause mortality. Complementary, the incidence of SAEs in randomised controlled trials (RCTs) with withdrawal and parallel designs was performed by meta-analysis.Results:A total of 31 observational studies were included (6811 patients totalizing 17530 patients-years [PY] of follow-up). The incidence rate of SAEs was similar in observational cohorts and withdrawal RCTs (4.46 events per 100 PY, 95% CI 2.85- 6.38, I2= 95%) and 3.71 events per 100 PY (95%CI 0.0-13.34), I2= 56%, respectively). The incidence of SAE was lower in parallel RCT. The incidence rate of serious infections, malignancies and death in observational cohorts was estimated at 0.74 events per 100 PY (95%CI 0.32-1.30, I2=83%), 0.10 events per 100 PY (95% CI 0.06-0.16, I2=0%) and 0.09 events per 100 PY (95% CI 0.05-0.14, I2=0%), respectively. Infections were the known cause of death in 8 of the 14 deaths. In meta-regression and subgroup analysis, variation of serious infections rates were partially explained by follow-up time (R2= 30.3%, p= 0.0008), JIA categories (all JIA versus polyarticular versus systemic JIA categories, p= 0.001) and cohort quality (Newcastle-Ottawa score ≥ to 6 versus ≤ to 5 stars, p= 0.0025).Conclusion:Our results suggest that the incidence rate of SAEs related to BAs in JIA disease is similar to those observed in randomised withdrawal trials. The overall incidence remained low. However, unsatisfactory description of SAEs prevents analysis of hospitalisation causes. Infection and, to a lesser extent, cancer and death, explain only part of burden of BAs.References:[1]Berard RA, Laxer RM. Learning the hard way: clinical trials in juvenile idiopathic arthritis. Ann Rheum Dis. 2018;77(1):1–2.[2]Swart J, Giancane G, Horneff G, Magnusson B, Hofer M, Alexeeva Е, et al. Pharmacovigilance in juvenile idiopathic arthritis patients treated with biologic or synthetic drugs: combined data of more than 15,000 patients from Pharmachild and national registries. Arthritis Res Ther. 2018 27;20(1):285.[3]Monti S, Grosso V, Todoerti M, Caporali R. Randomized controlled trials and real-world data: differences and similarities to untangle literature data. Rheumatol Oxf Engl. 2018 01;57(57 Suppl 7):vii54–Disclosure of Interests:None declared

Background Rheumatoid Arthritis is a chronic rheumatic disease characterised by polyarticular inflammation. The rheumatoid factor is one of the most known prognostic markers, not only its presence, but also the levels. It also presents different isotypes (IgG, IgM, IgA), which can affect the course of the disease. Objectives To analyse the presence of different rheumatoid factor (RF) isotypes in Paraguayan patients with rheumatoid arthritis (RA) and to study their association with clinical and analytical characteristics. Methods Descriptive, cross-sectional study. A large number of clinical and serological variables were recorded. The anti-CCP 3.1 and Rheumatoid factor (RF) isotypes IgA, IgG, and IgM were measured in serum samples by ELISA (enzyme-linked immunosorbent assay) (NV Results 103 patients with RA were included, 86.4% were female, with a median age of onset of 44.7±13.6 years, and the mean disease duration was 7.13±7.03 years. The olygoarticular onset was the most frequent (46.6%). 13.7% were smokers. Extra-articular manifestations were present in 13.5%. The most frequent treatment was methotrexate (84.3%). Erosions were observed in 43.2% of patients. 28% were in remission of the disease measured by the DAS28 index. The average of HAQ was 0.47%±0.58. 91.3% had anti-CCP positive, the mean anti-CCP levels were 290.5±152.8 U/mL. RF isotypes was observed in 75.7%, 53.4% and 38.8% for IgM, IgA and IgG respectively. Mean levels were as follow, IgA 85.62±56.6 U/mL, IgM 96.7±30.9 U/mL, IgG 70.98±72.42 U/mL. 32% of the patients had 2 isotypes of RF, while 25.2% had the 3 isotypes. The 57.3% had ≥2 isotypes of RF. We did not find significant differences when comparing gender, age, disease duration, form of onset, extra-articular manifestations, smoking status, erosions, disease activity, HAQ, treatment, between the different RF isotypes, and levels, except in the presence of anti-CCP with the RF-IgM isotype (p Conclusions This is the first study of RF isotypes in Paraguayan patients with RA. The most frequent isotype of RF was IgM. More than 50% of patients had 2 or more RF isotypes. The majority of patients with positive RF had high levels of different isotypes, being the highest IgM. Disclosure of Interest None declared

Background The PANLAR-EOA (early-onset-arthritis) project includes panamericanrheumatologists to determine regional characteristics of patients with early onset arthritis. Objectives To describe the cohort of Paraguayan patients included in PANLAR-EOA project. Methods Longitudinal, prospective, multicentricstudy. Patients were included according to the PANLAR-EOA project and registered in REPANARC(www.panlareoa.org) database. At baseline and annual visits, a large number of demographic, clinical and analytical variables were recorded. Quantitative variables were characterised by their means and standard deviations, while the qualitative variables were characterised according to the percentage of patients. The comparison of epidemiological and clinical variables was performed using the chi-squared test and the Wilcoxon test respectively for qualitative and quantitative variables, respectively. Results 136 patients with early onset arthritis were included, out of which 88 completed the 12 months follow up and 58 the 24 months one. In these, 86%were female with a median age of 43.9±13.2 years. The most frequent race was mestiza in 80.1%. According to GRAFFAR index, middle class was the predominant social stratum (9.8±3.1). The average number of years of schooling was 12.8±3.8. Polyarticular onset was registeredin 61% patients. During follow-up, 43.1% had positive rheumatoid factor and 56.5% positive anti-CCP. The diagnostic delay was 3.9±3.0 months. Initially, 63.2% (86/136) were diagnosed with rheumatoid arthritis (RA) and 36.8% (50/136) with undifferentiated arthritis (UA). The most frequent treatment was methotrexate (85.3%, 90.9%, and 89.3% at baseline, 1 and 2 years of follow-up respectively). During follow-up, a significant diagnostic change was observed in patients with UA (p=0.004, OR=2.9 [95%CI, 1.4–6.5]). The variables associated with RA diagnosis werepresence of anti-CCP (p=0.000, OR=15.8 [95%CI,5.4–51.1]), rheumatoid factor (p=0.000, OR=9.2 [95%CI,3.4–27.0]), smoking (p=0.032, OR=8.8 [95%CI,1.1–404.7]), high body mass index (p=0.041, OR=1.94 [95%CI,−0.2–4.1]) and high activity measured by the DAS28 index (p=0.01). After one year of follow-up there was a significant decrease in disease activity according to DAS 28 (p=2.2e-09[95% CI, −1.5,–0.9]), SDAI(p=1.2e-11[95% CI, 18.2,–11.2]) and HAQ (p=7.2e-08[95% CI,−0.7,–0.4]). Similar results were found at the 2nd year of follow-up, DAS28 (p=8.8e-06[95% CI,−1.6,–0.7]), SDAI (p=2.1 e- 07 [95% CI,−20.0,−10.3]) and HAQ (p=3.4e-08[95% CI,−0.9,–0.5]). Conclusions In this cohort of early onset arthritis, diagnostic delay was lower than that observed in other series and the rate of change from diagnosis of UA to RA was statistically significant during the first year of follow-up. A good control of the inflammatory activity of the disease was observed, with a significant improvement of all the variables analysed during its evolution. Disclosure of Interest None declared

Macroporous polypropylene (MPP) membranes incorporating poly[sodium (styrene sulphonate)], P(SSNa), and poly[(ar-vinylbenzyl) trimethylammonium chloride],P(ClVBTA) were modified via “in situ” radical polymerization. Modified and unmodified MPP were characterized using SEM and charge density properties. Donnan dialysis was used to transport chromium ions. The results show that the degree of modification varied between 2.5% and 4.0%, and the water uptake percentage varied between 15% and 20%. Experimental data for chromium ion transport (Cr(III) and Cr(VI)) were fitted to a mathematical model, and a correlation coefficient very close to 1 was obtained. The main parameters of this mathematical fit are k and a. These parameters report the concentration of chromium ions that converge in the Donnan equilibrium (k) and the response rate of the modified membrane (a).

The two co-authors of the mentioned above article were incorrect. The correct are authors should have been "P. A. Beltrán" instead of "P. A. B. Roa" and "J. F. Diaz-Coto" instead of "L. Diaz Soto".

The antimicrobial activity of different extracts of Alternanthera caracasana. HBK against 11 bacterial strains and 1 yeast strain was evaluated. The ethyl acetate extract showed antimicrobial activity against Staphylococcus aureus., Staphylococcus epidermidis., Bacillus subtilis., Sarcina lutea., and one strain of Vibrio cholerae.. There was no antimicrobial activity against Candida albicans.. As a bioactive compound, 7-methoxycoumarin was identified.

Background Systemic Lupus Erythematosus (SLE) is a systemic inflammatory disease associated with genetic, environmental, hormonal and immunological factors. Vitamin D levels are nowadays considered as one possible factor associated with disease activity. Therefore, previous studies have analyzed vitamin D to the severity of SLE. Objectives To assess the Vitamin D status in paraguayan SLE patients and its association with disease activity. Methods An observational Trial has been performed on individuals diagnosed with SLE. Epidemiological, clinical and biochemical data have been recorded for each patient to study the association between vitamin D concentrations, the phospho-calcium metabolism parameters and disease activity. Quantitative determination of Vitamin D was perform using chemoluminescence ARCHITEC assay. Vitamin D status was interpreted as follows: deficiency ≤20 ng/ml and insufficiency 21–29 ng/ml. The statistical association tests were performed using linear (SLEDAI activity index) and logistic (Inactive/Mild vs Moderate/Severe) regressions. The epidemiological, clinical and biochemical variables were used as explanatory variables in these models.This study is a preliminary analysis of a trial supported by CONACYT (Paraguay) to investigate the role of vitamin D in patients diagnosed with SLE. Results We included 77 SLE patients, of whom 94.8% (73/77) were female. The average age of patients at the time of the study was 30.7±10.3 years. All patients received calcium supplements associated with vitamin D. The average vitamin D concentration was 32.2±12.10 ng /ml. 29.9% (23/77) of patients had vitamin D insufficiency and 13.0% had vitamin D deficiency. 94.8% (73/77) of the population had normal serum calcium and the total population had a normal phosphoremia. As for the dosage of PTH, it was found that 27.3% (21/77) have high values of PTH. 20.8% (16/77) of the patients had positive anti-DNA. Low C3 complement was observed in 30/77 (39%) and low C4 in 50/77 (64.9%) patients. The mean value of SLEDAI at the time of the study was 2.32±2.83. When we study the distribution of vitamin D concentration according to the disease activity (SLEDAI) a clear pattern is observed linking lower vitamin D concentrations with higher disease activity (Figure 1). Patients with lower vitamin D concentrations are more likely to have higher disease activity (OR 0.93, 95% CI 0.88–0.99;P-Value=0.059. The means and standard deviations of vitamin D depending on the SLEDAI activity index are provided in Table 1. Conclusions In this preliminary study of Paraguayan SLE patients, Vitamin D deficiency was frequent despite treatment with supplements. In addition, a clear association between SLEDAI and Vitamin D values was observed. The final analysis in a larger patient cohort will have to confirm these findings and clarify relation with disease activity. References Eloi M, Horvath DV, Ortega JC, Prado MS, Andrade LE, Szejnfeld VL, de Moura Castro CH. 25-Hydroxivitamin D Serum Concentration, Not Free and Bioavailable Vitamin D, Is Associated with Disease Activity in Systemic Lupus Erythematosus Patients. PLoS One. 2017 Jan 13;12(1). Disclosure of Interest None declared

RESUMO: O objetivo do estudo foi avaliar as variáveis hemogasométricas de cordeiros nascidos a termo e prematuros do nascimento às 48 horas de vida. Foram constituídos quatros grupos experimentais: PN (cordeiros nascidos de parto normal, n=15, média de 146 dias de gestação); PNDEX (cordeiros nascidos de parto normal, cujas mães receberam 16 mg de dexametasona aos 141 de gestação, n=8, média de 143 dias de gestação); PRE (cordeiros prematuros nascidos de cesarianas aos 138 dias de gestação, n=10) e PREDEX (cordeiros prematuros nascidos de cesarianas aos 138 dias de gestação, cujas mães receberam 16 mg de dexametasona dois dias antes do parto, n=9). Imediatamente após o nascimento, os cordeiros de todos os grupos apresentaram quadro de acidose respiratória (pH baixo e pCO2 elevada), com maior ênfase nos animais prematuros. A concentração de HCO3 - diminuiu entre 15 e 60 minutos de vida, principalmente nos grupos PRE e PREDEX, com posterior aumento no M24h. Os valores de diferença de base foram menores nos cordeiros prematuros, os quais apresentaram respiração abdominal, intensa dispneia e grande quantidade de líquido pulmonar. A estabilização do equilíbrio ácidobase ocorreu em todos os animais ao longo das primeiras 24 horas de vida. A dexametasona teve influência positiva sobre a condição clínica dos animais prematuros, resultando em adequada ventilação e perfusão tecidual, o que garantiu maior taxa de sobrevivência. ABSTRACT: The aim of the study was to evaluate blood gas parameters of full-term and premature lambs from birth to 48 hours of life. Four experimental groups were formed: NDG (normal delivery group - lambs vaginally delivered, n=15, average of 146-day gestation); NDEXG (normal delivery with dexamethasone group - lambs vaginally delivered whose mothers received 16 mg of dexamethasone at 141 days of gestation, n=8, average of 143-day gestation); PRE (premature lambs born by cesarean section at 138 days of gestation, n=10) and PREDEX (premature lambs born by cesarean section at 138 days gestation, whose mothers received 16mg of dexamethasone two days before, n=9). Immediately after birth, lambs from all groups showed respiratory acidosis (low pH and high pCO2), most obviously in premature animals. The concentration of HCO3 - was lower between 15 and 60 minutes of life, especially in PRE and PREDEX groups with subsequent increase in M24h. The values of base excess were lower in premature lambs, which showed abdominal breathing, severe dyspnea and lots of lung fluid. The stabilization of acid-base balance occurred in all animals during the first 24 hours of life. Dexamethasone had a positive effect on the clinical condition of the premature lambs, resulting in adequate ventilation and tissue perfusion, which guaranteed higher survival rate.

Made available in DSpace on 2018-12-11T17:24:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-01-01 Made available in DSpace on 2014-12-03T13:08:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-06-01Bitstream added on 2014-12-03T13:24:41Z : No. of bitstreams: 1 S0103-84782014000600025.pdf: 135978 bytes, checksum: b8668a6c4489ae5ca6a3ec71952cf8a9 (MD5) Item merged in doublecheck by Luana Priscila Costa (luana@reitoria.unesp.br) on 2019-04-26T18:53:50Z Item was identical to item(s): 179488, 110581 at handle(s): http://hdl.handle.net/11449/177208, http://hdl.handle.net/11449/111481 O objetivo deste trabalho foi avaliar a influência do glicocorticoide sobre parâmetros vitais de cordeiros nascidos a termo e prematuros, do nascimento às 48 horas de vida. Foram constituídos quatros grupos: PN (cordeiros nascidos de parto normal, n=15, média de 146 dias); PNDEX (cordeiros nascidos de parto normal, cujas mães receberam 16mg de dexametasona aos 141 dias de gestação, n=8, média de 143 dias); PRE (cordeiros prematuros nascidos de cesarianas aos 138 dias de gestação, n=10); e PREDEX (cordeiros prematuros nascidos de cesarianas aos 138 dias de gestação, cujas mães receberam 16mg de dexametasona dois dias antes, n=9). As frequências cardíaca e respiratória variaram ao longo do período, com os maiores valores nos grupos de partos normais. A temperatura retal diminuiu em todos os grupos nos primeiros 60 minutos de vida, sendo os menores valores observados nos cordeiros prematuros, e o escore Apgar foi mais alto nos animais nascidos em tempo gestacional normal. Os cordeiros prematuros apresentaram menor vitalidade e menor taxa de sobrevivência, entretanto, menor taxa de mortalidade foi observada nos prematuros sob influência da dexametasona. The aim of the study was to evaluate the influence of glucocorticoids on vital parameters of full-term and preterm lambs from birth to 48 hours of life. Four experimental groups were formed: NDG (normal delivery group - lambs vaginally delivered, n=15, average of 146-day gestation); NDEXG (normal delivery with dexamethasone group - lambs vaginally delivered whose mothers received 16mg of dexamethasone at 141 days of gestation, n=8, average of 143-day gestation); PRE (premature lambs born by cesarean section at 138 days of gestation, n=10) and PREDEX (premature lambs born by cesarean section at 138 days gestation, whose mothers received 16mg of dexamethasone two days before, n=9). Heart and respiratory rates had variations during the observation period, with the highest mean values in the groups of normal deliveries (NDG and NDEXG). Rectal temperature decreased in all groups in the first 60 minutes of life, with the lowest mean values in premature lambs (PRE and PREDEX) and the Apgar score was higher in animals delivered at normal gestational time. Preterm lambs had lower vitality and chance of survival, however, lower mortality rate was observed in offspring of ewes that received dexamethasone two days before surgery. Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Rua Clóvis Pestana, 793, 16050-680, Araçatuba, SP Universidade Federal de Mato Grosso do Sul (UFMS), Campo Grande, MS Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Araçatuba, SP Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Rua Clóvis Pestana, 793, 16050-680, Araçatuba, SP Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Araçatuba, SP

Background Cancer is a high-impact disease throughout the world. A negative correlation has been established between the development of cancer and the Th2 immune response. Infection by helminth parasites is characterized by the induction of a strong and long-lasting Th2 response. The aim of this work was to evaluate the effect of the immune response induced by the infection with the helminth Hymenolepis nana, on the tumorigenesis induced by dimethylbenz-anthracene (DMBA) in mice. Methodology Four different groups of 14 female BALB/c mice were formed; Group A, dimethyl sulfoxide (DMSO) (vehicle) was administered cutaneously, Group B infected with H. nana , group C, cutaneously DMBA and finally Group D infected with H. nana and cutaneous DMBA. The tumor load was determined in those animals that developed cancerous lesions. In all groups were determined: serum concentration of IgE, IFNγ, IL-10, IL-5 and malondialdehyde (MDA). The inflammatory infiltrate was analyzed from skin samples and the expression of the main eosinophilic protein and myeloperoxidase was determined. Results The group previously infected with H. nana had a reduced amount of tumors with smaller size, in comparison to the group that received only DMBA; this reduction was associated with lower levels of IFNγ and IL-10, while levels of IgE, IL-5 and MDA were higher. Further, the number of eosinophils and neutrophils was statistically higher in the animals that were previously infected with the helminth and developed less tumors. Conclusion The immune response induced by H. nana infection is associated with the reduction of tumors probably due to the activity of eosinophils and neutrophils.

The effects of phytochemicals occurred in fractions and extracts of fruits of “Maqui-berry” (Aristotelia chilensis), on the expression of cyclooxygenase-2 (COX-2), inducible-nitric oxide synthases (iNOS) and the production of proinflammatory mediators were investigated in lipopolysaccharide (LPS)-activated murine macrophage RAW-264 cells, as well as their antioxidant activities. The MeOH extract (A), acetone/methanol extract (B), fractions F3, F4, subfractions (SF4-SF6, SF7, SF8-SF10, SF11-SF15, SF16-SF20), quercetin, gallic acid, luteolin, myricetin, mixtures M1, M2 and M3 exhibited potent anti-inflammatory and antioxidant activities. The results indicated that anthocyanins, flavonoids and its mixtures suppressed the LPS induced production of nitric oxide (NO), through the down-regulation of iNOS and COX-2 protein expressions and showed a potent antioxidant activity against SOD, ABTS, TBARS, ORAC, FRAP and DCFH. The inhibition of enzymes and NO production by selected fractions and compounds was dose-dependent with significant effects seen at concentration as low as 1.0–50.0 (ppm) and 5.0–10.0 μM, for samples (extracts, fractions, subfractions and mixtures) and pure compounds, respectively. Thus, the phenolics (anthocyanins, flavonoids, and organic acids) as the fractions and mixtures may provide a potential therapeutic approach for inflammation associated disorders and therefore might be used as antagonizing agents to ameliorate the effects of oxidative stress.

Type 1 diabetes (also known as juvenile diabetes) results from the autoimmune destruction of insulin-producing pancreatic β cells, and survival of individuals with the disease depends mainly on daily insulin injections or the use of insulin pumps. Transplantation of donor islets has recently proven effective at restoring insulin production and blood glucose control in type 1 diabetes mellitus patients (1–4). Its widespread application is, however, limited by availability of donor islets, the requirement for long-term use of immunosuppressive drugs to prevent graft rejection, and lower rates of long-term insulin independence (5). Moreover, the reduced islet viability and loss of insulin content that occurs during islet isolation and pre-transplantation culture necessitates the use of large numbers of islets, usually from multiple donors, to achieve insulin independence (6). Identification of factors that can enhance islet survival in vitro and improve islet function post-transplantation will thus help in extending the period of insulin independence in type 1 diabetes patients and reduce the number of donors required for successful islet transplantation. Glial cell line-derived neurotrophic factor (GDNF) is a factor produced by glial cells that plays an important role in the development of the enteric nervous system. GDNF has previously been shown to promote the survival of β-cells in vitro, and to enhance β-cell mass and improve glucose control in mice when transgenically over-expressed under the control of the glial fibrillary acidic protein promoter (2, 7). In this study we investigated the ability of GDNF to improve human islet post-transplant survival and function. Using in vitro and in vivo methods we assessed the effects of culturing human islets in the presence of GDNF on β-cell survival, in vitro insulin secretion, and post transplantation glycemic control. We performed these studies to assess the potential for use of GDNF to enhance human islet viability before transplant in type 1 diabetes patients.

Outcomes in transplantation have been limited by suboptimal long-term graft survival and toxicities associated with current immunosuppressive approaches. T cell costimulation blockade has shown promise as an alternative strategy to avoid the side effects of conventional immunosuppressive therapies, but targeting CD28-mediated costimulation alone has proven insufficient to prevent graft rejection in primates. Donor-specific memory T (TM) cells have been implicated in costimulation blockade-resistant transplant rejection, due to their enhanced effector function and decreased reliance on costimulatory signaling. Thus, we have tested a potential strategy to overcome TM cell-driven rejection by targeting molecules preferentially expressed on these cells, such as the adhesion molecule lymphocyte function-associated antigen 1 (LFA-1). Here, we show that short-term treatment (i.e., induction therapy) with the LFA-1-specific antibody TS-1/22 in combination with either basiliximab (an IL-2Rα-specific mAb) and sirolimus (a mammalian target of rapamycin inhibitor) or belatacept (a high-affinity variant of the CD28 costimulation-blocker CTLA4Ig) prolonged islet allograft survival in nonhuman primates relative to control treatments. Moreover, TS-1/22 masked LFA-1 on TM cells in vivo and inhibited the generation of alloproliferative and cytokine-producing effector T cells that expressed high levels of LFA-1 in vitro. These results support the use of LFA-1-specific induction therapy to neutralize costimulation blockade-resistant populations of T cells and further evaluation of LFA-1-specific therapeutics for use in transplantation.

Penstemon gentianoides (Kunth) Poir. and Penstemon campanulatus (Cav.) Willd. (Plantaginaceae) are important medicinal plants in Mexico used by indigenous people for their anti-inflammatory effects and to also reduce rheumatic pains.In addition to radical scavenging activity, the anti-inflammatory activity of the extracts, fractions and compounds of these plants were investigated and reported here for the first time.The anti-inflammatory activities of MeOH, CH(2)Cl(2), and ethyl acetate extracts and iridoid, flavonoids, and phenylpropanoids from Penstemon gentianoides and P. campanulatus were studied in the TPA-induced mouse ear edema model. In addition, antioxidant activity against DPPH, crocin and β-carotene were investigated.All extracts were tested and a selection of known compounds significantly (p0.05) inhibited mouse ear edema. The results showed that CH(2)Cl(2) extracts of roots and stems from P. gentianoides and ethyl acetate extracts of leaves from P. gentianoides and P. campanulatus, as well as luteolin, diosmetin, penstemide and verbascoside produced the most positive results. Of all substances tested, the CH(2)Cl(2) extract of P. gentianoides roots was the most powerful inhibitor (ED(50)=0.07 mg/ear), with activity comparable to that of indomethacin. These extracts, compounds purified, as well as known compounds, inhibited oxidation of β-carotene and crocin.These findings showed that the iridoid monoterpenes, flavonoids and phenylpropanoids present in these plants species may all contribute to the observed anti-inflammatory activity. Additionally, the observed antioxidant activity is correlated with the anti-inflammatory activity of these plants and the phytochemicals derived from them.

PD-1 immune checkpoint blockade occasionally results in durable clinical responses in advanced metastatic cancers. However, mechanism-based predictors of response to this immunotherapy remain incompletely characterized. We performed comprehensive genomic profiling on a tumor and germline sample from a patient with refractory lung adenocarcinoma who achieved marked long-term clinical benefit from anti–PD-L1 therapy. We discovered activating somatic and germline amino acid variants in JAK3 that promoted PD-L1 induction in lung cancer cells and in the tumor immune microenvironment. These findings suggest that genomic alterations that deregulate cytokine receptor signal transduction could contribute to PD-L1 activation and engagement of the PD-1 immune checkpoint in lung cancer. Cancer Immunol Res; 3(8); 855–63. ©2015 AACR.

The antimicrobial activity of different extracts of Tagetes lucida. Cav. (Asteraceae) against 11 bacterial strains and one yeast strain (Candida albicans.) was evaluated. The ethyl acetate extract showed antibacterial activity against Shigella boydii., Staphylococcus aureus., Staphylococcus epidermidis., Pseudomonas aeruginosa., Bacillus subtilis., Sarcina lutea., and four strains of Vibrio cholerae.. The bioactive compound 5,7,4′-trimethoxyflavone was identified.

The success of the Edmonton Protocol for islet transplantation has provided new hope in the treatment of type 1 diabetes. This study reports on the assessment of 83 human islet grafts transplanted using the Edmonton Protocol since 1999. Cellular composition, as assessed by immunohistochemistry, showed a lower islet purity (∼40%) than has been reported in previous studies using dithizone staining to quantitate islet equivalents. Furthermore, grafts were found to contain substantial populations of exocrine and ductal tissue. Total cellular insulin transplanted was 8,097.6 ± 3,164.4 μg/patient, and was significantly lower in bottom gradient layer grafts than top gradient layer or whole/combined grafts (P < 0.0005). A static incubation test for islet function gave a stimulation index of 3–4, although this measure did not correlate with posttransplant metabolic outcome. Furthermore, we confirmed a previously reported trend in which donor age affects islet yield and purity. It is important to note that a significant positive correlation was observed between the number of islet progenitor (ductal-epithelial) cells transplanted and long-term metabolic success as assessed an by intravenous glucose tolerance test at ∼2 years posttransplant. In summary, careful assessment of islet graft composition is needed in a clinical transplantation program to accurately estimate islet purity and assess the contribution of other cell types present, such as islet progenitor cells.

Human pancreas preservation for islet transplantation holds additional challenges and considerations compared with whole pancreas transplantation. The purpose of this study was to clarify the limitations of the University of Wisconsin (UW) solution and the potentials of the two-layer method (TLM) for pancreas preservation before human islet isolation.We retrospectively evaluated human islet isolation records between January 2001 and February 2003. One hundred forty-two human pancreata were procured from cadaveric donors and preserved by means of the UW solution (n=112) or TLM (n=30). Human islet isolations were performed using a standard protocol and assessed by islet recovery and in vitro function of islets.Eight to ten hours of cold ischemia in the UW solution is a critical point for successful islet isolations. It is difficult to recover a sufficient number of viable islets for transplantation from human pancreata with more than 10 hours of cold storage in the UW solution. The overall islet recovery in the TLM group was significantly higher than in the UW group. With 10 to 16 hours of cold storage, the success rates of islet isolations remained at 62% in the TLM group but decreased to 22% in the UW group. Transplanted islets in the TLM group worked well in the recipients.There are time limitations for using the UW solution for pancreas preservation before human islet isolation. The TLM is a potential method to prolong the optimal cold storage time for successful islet isolations.

BACKGROUND A major obstacle to successful small bowel transplantation is that of bacterial infection. The aim of this study was to preserve the small bowel mucosal barrier by using oxygenated luminal perfusion with a proven amino acid (AA)-based solution. METHODS Rat small bowel (n=4) was flushed vascularly with modified University of Wisconsin solution and flushed luminally as follows: group 1, none (control); group 2, AA solution; group 3, 1-hr perfusion then storage with AA; group 4, continuous perfusion with AA. Energetics, malondialdehyde (MDA), glutathione (reduced), and histology were assessed over 24 hr at 4 degrees C. RESULTS Within 4 hr, adenosine triphosphate (ATP) dropped by 25% to 65% in all groups except for group 4, which remained unchanged from fresh tissue values throughout 12 hr. After 12 hr, ATP in groups 1 through 3 had dropped to 0.5 to 0.9 micromol/g, compared with 1.5 micromol/g for group 4. Even after 24 hr, group 4 levels were more than twofold greater than groups 1 through 3. MDA increased transiently in tissues subjected to simple flush (no perfusion), whereas levels in perfused tissues remained elevated throughout the 24-hr period. Glutathione in group 1 dropped by greater than 50% from fresh tissue values but increased over 24 hr in groups 2 and 3 by 50% to 55%. Overall, histologic injury was markedly less in groups 2 through 4; however, after 24 hr, the lowest injury was observed in group 3 (median, grade 2) compared with groups 1 and 4 (grades 7 and 4). CONCLUSIONS Our data indicate that perfusion clearly improves tissue energetics. However, mucosal integrity is markedly superior, with only a brief 1-hr period of perfusion; oxidative and mechanical stress are the factors likely responsible for injury resulting from continuous perfusion.

BACKGROUND This study improved small bowel preservation using University of Wisconsin (UW) solution in conjunction with hypothermic luminal perfusion. METHODS Small bowels from Sprague-Dawley rats (n=4) were flushed vascularly with modified UW solution and flushed luminally: group 1, none (clinical control); group 2, UW solution; group 3, 1-hr oxygenated perfusion then static storage with UW; and group 4, 24-hr continuous oxygenated perfusion with UW. Energetics, lipid peroxidation, and histology were assessed during 24 hr at 4 degrees C. RESULTS After 12 hr, adenosine triphosphate ranged from 0.5 to 0.8 micromol/g in groups 1 to 3 compared with 1.5 micromol/g in group 4. Even after 24 hr, levels in group 4 were more than twofold greater than levels in groups 1 to 3. Energy charge values ([adenosine triphosphate+adenosine diphosphate/2]/total adenylates) decreased from fresh tissue values of 0.69 in all groups except group 4 throughout 24-hr perfusion. Malondialdehyde (MDA; a product of lipid peroxidation) doubled within 4 hr in group 1 and remained high throughout storage. In groups 3 and 4, MDA levels increased as the time of perfusion increased; group 2 showed no elevated MDA levels at any time. After 12 hr, histologic integrity was superior in groups 3 and 4; however after 24 hr, the best Park's grade was observed in group 3 (median grade 4) compared with groups 1 (grade 7) and 4 (grade 6). CONCLUSIONS Our data indicate that perfusion clearly improves tissue energetics; however, mucosal integrity is superior with only a brief 1-hr period of luminal perfusion, despite limited improvements in energetics.

BACKGROUND A two-layer (University of Wisconsin solution/perfluorochemical [UW/PFC]) cold-storage method delivers sufficient oxygen to the pancreas during preservation and restores the ischemically damaged pancreas. In this study, we determined whether the additional preservation by the two-layer method could improve islet recovery from human pancreases with prolonged cold storage in UW. METHODS Human pancreases were procured from cadaveric organ donors and preserved by the two-layer method (UW/PFC) for 2.9+/-0.7 hours (mean+/-SEM) at 4 degrees C after 11.8+/-1.5 hours of cold storage in UW (UW/PFC group, n=7), or by cold UW alone for 11.3+/-0.3 hours (UW group, n=14). The selected pancreases met the criteria of having at least 10 hours of cold storage in UW. All were processed by using a standard protocol of Liberase perfusion with Pefabloc by way of the duct, gentle mechanical dissociation, and Ficoll gradient purification. Transplanted islets were selected with the criteria of the Edmonton protocol (>5,000 islet equivalents [IE]/kg recipient body weight). RESULTS The islet recovery was significantly increased in the UW/PFC group compared with the UW group (349.2+/-44.1 x 10 and 214.0+/-31.0 x 10 IE, respectively

In vitro proliferation of isolated pancreaticislets has become an area of great interest given the scarcity of clinicalisletdonors and the islet mass requirements for clinical islet transplantation.Smallintestinal submucosa (SIS), a naturally occurring extracellular matrix, hasbeeninvestigated to promote wound healing, tissue remodeling and cell growth. Thisstudy evaluated recovery and function of isolated canine pancreatic isletsfollowing in vitro tissue culture. Pancreatic islets wereisolated from mongrel dogs using standard surgical procurement followed byintraductal collagenase distension, mechanical dissociation and EuroFicollpurification. Groups of purified islets were cultured in a humidifiedatmosphereof 95% air and 5% CO2 for 48 hours in standard islet cultureconditions of CMRL 1066 tissue culture media (Gibco) which had beensupplementedwith 25μM HEPES, penicillin/streptomycin and either 10% heat inactivatedfetal calf serum (FCS, Gibco) or solubilized SIS solution (Cook Biotech, Inc.,West Lafayette, IN). The mean recovery of islets following the culture periodwas determined by sizing duplicate counts of a known volume and viability wasassessed by static incubation with low glucose (2.8 mM), highglucose (20 mM) and high glucose solution supplemented with 50μm IBMX solution. Remaining islets were embeddedhistologically.From a consecutive series of six culture experiments, a significantly higher (p 150 μm increasedfrom 24% to 31% in the SIS-treated group, whereas the same proportion decreasedto 18% from 22% in the control (FCS-treated) group. Histological evaluation offixed tissue samples collected following the culture period identified insulinand glucagon-secreting cells in the SIS and FCS treated groups, however ahigherfrequency of insulin positive cells were detected consistently in the SIStreated group. A proliferation marker (PCNA) identified positive cells withinboth groups as well. This study suggests that co-culture of freshly isolatedcanine islets in medium supplemented with solubilized SIS can improve thepost-culture recovery and in vitro islet function. Futureinvestigations will focus on the cellular interactions of SIS, bothinvitro and in vivo.

We evaluate the mode of action of verbascoside obtained from Buddleja cordata against Staphylococcus aureus by killing kinetics and incorporation of precursors methods. Verbascoside induced lethal effect on S. aureus, by affecting protein synthesis and inhibiting leucine incorporation.

Background Systemic Lupus Erythematosus (SLE) and rheumatoid arthritis (RA) are autoimmune diseases that mainly affect people in the reproductive stage of their life; it is interesting to investigate the impact of both diseases have in patient9s quality of life. Objectives The main objective of this study was to evaluate the quality of life of patients diagnosed with SLE and RA using the SF36 questionnaire and the factors that influence it Methods Analytical, observational cross-sectional study of a cohort of SLE and RA patients. A large number of clinical and epidemiological variables were recorded (i.e. years of evolution, the age at diagnosis, disease activity, gender, marital status, etc). Quality of life was determined by SF36 questionnaire. Statistical analysis was performed with SPSS19.0 program. Qualitative variables were expressed as percentages and frequencies while quantitative were informed as mean and standard deviation. Association was analyzed with X2 test and Student T test. Pearson9s test was used for correlation analysis. Results 112 patients were studied, 50.9% (57/112) patients had SLE and 49.1% (55/112) RA. 69.6% (78/112) of patients came from the capital city and metropolitan area. 87.5% (90/112) were female, 57.14% (64/112) were living in couple and 49.1% (55/100) had basic education. 43.8% (49/97) perceive less or equal to the minimum wage monthly income. 42% (47/98) were living with 3 or fewer inhabitants in the house and 58.9% (66/108) had ≥40 years old at disease onset. The mean value of physical (PCS) component of SF36 questionnaire in SLE and RA was 45.13±12.5 and 47.7±12.5 respectively and the difference between them was significantly (p=0.01). On the other hand, the mean value of mental (MCS) component was 47.7±12.5 and 46.4±12 respectively but no significant difference was found (p=0.58). In relation to the 8 SF36 components we found significant difference between them, principally in the components that measures body pain (p=0.0001) and health perception (p=0.047). In SLE patients, a significant negative correlation it was found between PCS and HAQ (p=0.00018) and SLEDAI (p=0.0020). No significantly correlation with MCS was found. Regarding the SLICC Score, a statistically negative correlation was found with MCS (p=0.036). In RA patients, a significant negative correlation was found between PCS and HAQ (p=0.0001), no association with the DAS28 was found. But, a negative correlation between the MCS and DAS28 (p=0.03) was found. No significant correlation was observed with HAQ. Conclusions In this cohort revealed that physical and mental component of SF36 are influenced by the disease activity and functional impairment. Acknowledgement Sr Horacio Medina Disclosure of Interest None declared

Immunosuppressive therapies that block the CD40/CD154 costimulatory pathway have proven to be uniquely effective in preclinical xenotransplant models. Given the challenges facing clinical translation of CD40/CD154 pathway blockade, we examined the efficacy and tolerability of CD40/CD154 pathway-sparing immunomodulatory strategies in a pig-to-nonhuman primate islet xenotransplant model. Rhesus macaques were rendered diabetic with streptozocin and given an intraportal infusion of ≈ 50 000 islet equivalents/kg wild-type neonatal porcine islets. Base immunosuppression for all recipients included maintenance therapy with belatacept and mycophenolate mofetil plus induction with basiliximab and LFA-1 blockade. Cohort 1 recipients (n = 3) were treated with the base regimen alone; cohort 2 recipients (n = 5) were additionally treated with tacrolimus induction and cohort 3 recipients (n = 5) were treated with alefacept in place of basiliximab, and more intense LFA-1 blockade. Three of five recipients in both cohorts 2 and 3 achieved sustained insulin-independent normoglycemia (median rejection-free survivals 60 and 111 days, respectively), compared to zero of three recipients in cohort 1. These data show that CD40/CD154 pathway-sparing regimens can promote xenoislet survival. Further optimization of these strategies is warranted to aid the clinical translation of islet xenotransplantation.

Islet transplantation to treat type 1 diabetes has been limited in part by toxicities of current immunosuppression and recipient humoral sensitization. Blockade of the CD28/CD80/86 and CD40/CD154 pathways has shown promise to remedy both these limitations, but translation has been hampered by difficulties in translating CD154-directed therapies. Prior CD40-directed regimens have led to prolonged islet survival, but fail to prevent humoral allosensitization. We therefore evaluated the addition of CTLA4Ig to a CD40 blockade-based regimen in nonhuman primate (NHP) alloislet transplantation. Diabetic rhesus macaques were transplanted allogeneic islets using the CD40-specific antibody 3A8, basiliximab induction, and sirolimus with or without CTLA4Ig maintenance therapy. Allograft survival was determined by fasting blood glucose levels and flow cytometric techniques were used to test for donor-specific antibody (DSA) formation. CTLA4Ig plus 3A8, basiliximab and sirolimus was well tolerated and induced long-term islet allograft survival. The addition of CTLA4Ig prevented DSA formation, but did not facilitate withdrawal of the 3A8-based regimen. Thus, CTLA4Ig combines with a CD40-specific regimen to prevent DSA formation in NHPs, and offers a potentially translatable calcineurin inhibitor-free protocol inclusive of a single investigational agent for use in clinical islet transplantation without relying upon CD154 blockade.

Costimulation blockade of the CD40/CD154 pathway has been effective at preventing allograft rejection in numerous transplantation models. This strategy has largely depended on mAbs directed against CD154, limiting the potential for translation due to its association with thromboembolic events. Though targeting CD40 as an alternative to CD154 has been successful at preventing allograft rejection in preclinical models, there have been no reports on the effects of CD40-specific agents in human transplant recipients. This delay in clinical translation may in part be explained by the presence of cellular depletion with many CD40-specific mAbs. As such, the optimal biologic properties of CD40-directed immunotherapy remain to be determined. In this report, we have characterized 3A8, a human CD40-specific mAb and evaluated its efficacy in a rhesus macaque model of islet cell transplantation. Despite partially agonistic properties and the inability to block CD40 binding of soluble CD154 (sCD154) in vitro, 3A8-based therapy markedly prolonged islet allograft survival without depleting B cells. Our results indicate that the allograft-protective effects of CD40-directed costimulation blockade do not require sCD154 blockade, complete antagonism or cellular depletion, and serve to support and guide the continued development of CD40-specific agents for clinical translation.

Significant deficiencies in understanding of xenospecific immunity have impeded the success of preclinical trials in xenoislet transplantation. Although galactose-α1,3-galactose, the gal epitope, has emerged as the principal target of rejection in pig-to-primate models of solid organ transplant, the importance of gal-specific immunity in islet xenotransplant models has yet to be clearly demonstrated. Here, we directly compare the immunogenicity, survival and function of neonatal porcine islets (NPIs) from gal-expressing wild-type (WT) or gal-deficient galactosyl transferase knockout (GTKO) donors. Paired diabetic rhesus macaques were transplanted with either WT (n = 5) or GTKO (n = 5) NPIs. Recipient blood glucose, transaminase and serum xenoantibody levels were used to monitor response to transplant. Four of five GTKO versus one of five WT recipients achieved insulin-independent normoglycemia; transplantation of WT islets resulted in significantly greater transaminitis. The WT NPIs were more susceptible to antibody and complement binding and destruction in vitro. Our results confirm that gal is an important variable in xenoislet transplantation. The GTKO NPI recipients have improved rates of normoglycemia, likely due to decreased susceptibility of xenografts to innate immunity mediated by complement and preformed xenoantibody. Therefore, the use of GTKO donors is an important step toward improved consistency and interpretability of results in future xenoislet studies.

Human islet isolation outcomes were compared between two purification methods; 32 pancreases were processed by conventional Biocoll purification method (SM, standard method) and 132 pancreases by a refined University of Illinois at Chicago UW/Biocoll method (UIC-UB). There was no difference in donor characteristics between the two study groups. The prepurification equivalent islet number was similar between the groups (359,425+/-40,794 equivalent islet number in SM vs. 370,682+/-17,579 in UIC-UB). SM purified islets were mostly collected in only 2 of 12 fractions (68.9% and 36.3% purity). With the UIC-UB, highly purified islets were collected in 6 of 12 separate fractions (fractions 3-8 with purity of 84.8%, 82.5%, 72.0%, 59.3%, 46.8%, and 36.2%). UIC-UB yielded significantly greater islet yield compared with SM (368,419+/-18,245 vs. 260,908+/-37,835, P=0.017). Islet recovery rate was superior in UIC-UB (84.9% vs. 64.5%, P=0.04). Our study demonstrates a superior recovery of highly pure human pancreatic islets after purification using the refined method of UIC-UB gradient.

During isolation, islets are exposed to warm ischemia. In this study, intraductal administration of oxygenated polymerized, stroma-free hemoglobin-pyridoxalated (Poly SFH-P) was performed to improve O2 delivery. Rat pancreata subjected to 30-min warm ischemia were perfused intraductally with collagenase in oxygenated Poly SFH-P/RPMI or RPMI (control). PO2 was increased by Poly SFH-P (381.7 +/- 35.3 mmHg vs. 202.3 +/- 28.2, p = 0.01) and pH maintained within physiological range (7.4-7.2 vs. 7.1-6.6, p = 0.009). Islet viability (77% +/- 4.6 vs. 63% +/- 4.7, p = 0.04) was improved and apoptosis lower with Poly SFH-P (caspase-3: 34,714 +/- 2167 vs. 45,985 +/- 1382, respectively, p = 0.01). Poly SFH-P improved islet responsiveness to glucose as determined by increased intracellular Ca2+ levels and improved insulin secretion (SI 5.4 +/- 0.1 vs. 3.1 +/- 0.2, p = 0.03). Mitochondrial integrity was improved in Poly SFH-P-treated islets, which showed higher percentage change in membrane potential after glucose stimulation (14.7% +/- 1.8 vs. 9.8 +/- 1.4, respectively, p0.05). O2 delivery by Poly SFH-P did not increase oxidative stress (GSH 7.1 +/- 2.9 nm/mg protein for Poly SFH-P vs. 6.8 +/- 2.4 control, p = 0.9) or oxidative injury (MDA 1.8 +/- 0.9 nmol/mg protein vs. 6.2 +/- 2.4, p = 0.19). Time to reach normoglycemia in transplanted diabetic nude mice was shorter (1.8 +/- 0.4 vs. 7 +/- 2.5 days, p = 0.02), and glucose tolerance improved in the Poly SFH-P group (AUC 8106 +/- 590 vs. 10,863 +/- 946, p = 0.03). Oxygenated Poly SFH-P improves islet isolation and transplantation outcomes by preserving mitochondrial integrity.

The increasing incidence of diabetes and the need to further understand its cellular basis has resulted in the development of new diagnostic and therapeutic techniques. Nonetheless, the quest to noninvasively ascertain beta-cell mass and function has not been achieved. Manganese (Mn)-enhanced MRI is presented here as a tool to image beta-cell functionality in cell culture and isolated islets. Similar to calcium, extracellular Mn was taken up by glucose-activated beta-cells resulting in 200% increase in MRI contrast enhancement, versus nonactivated cells. Similarly, glucose-activated islets showed an increase in MRI contrast up to 45%. Although glucose-stimulated Ca influx was depressed in the presence of 100 microM Mn, no significant effect was seen at lower Mn concentrations. Moreover, islets exposed to Mn showed normal glucose sensitivity and insulin secretion. These results demonstrate a link between image contrast enhancement and beta-cell activation in vitro, and provide the basis for future noninvasive in vivo imaging of islet functionality and beta-cell mass.

Oxidative stress during islet isolation induces a cascade of events injuring islets and hampering islet engraftment. This study evaluated islet isolation and transplantation outcomes after intra-ductal glutamine administration. Human pancreata deemed unsuitable for pancreas or islet transplantation were treated with either a 5 mM solution of l-glutamine (n = 6) or collagenase enzyme alone (n = 6) through the main pancreatic duct. Islet yield, viability, in vitro function; markers of oxidative stress [malondialdehyde (MDA) and Glutathione (GSH)] and apoptosis were assessed. Islet yields were significantly increased in the glutamine group compared to controls (318, 559 +/- 25, 800 vs. 165, 582 +/- 39, 944 mean +/- SEM, p < 0.01). The amount of apoptotic cells per islet was smaller in the glutamine group than the control. The percentage of nude mice rendered normoglycemic with glutamine-treated islets was higher than the controls (83% n = 10/12 vs. 26% n = 6/23; p < 0.01), and the time to reach normoglycemia was decreased in the glutamine group (1.83 +/- 0.4 vs. 7.3 +/- 3 days; p < 0.01). Glutamine administration increased GSH levels (7.6 +/- 1.7 nmol/mg protein vs. 4.03 +/- 0.5 in control, p < 0.05) and reduced lipid-peroxidation (MDA 2.45 +/- 0.7 nmol/mg of protein vs. 6.54 +/- 1.7 in control; p < 0.05). We conclude that intra-ductal administration of glutamine reduces oxidative injury and apoptosis and improves islet yield and islet graft function after transplantation.

The aim of this study was to improve small bowel (SB) quality during cold storage by combining two proven preservation strategies involving perfluorocarbon (PFC) and a novel luminal amino acid-rich solution. Rodent SB was flushed vascularly with UW solution and flushed luminally as follows: Group 1 (control) – no luminal flush, stored in UW; Group 2 – luminal UW solution, stored in PFC; Group 3 – luminal amino-acid (AA) solution, stored in PFC; and Group 4 – luminal AA solution, stored in AA solution. Energetics, histology and mucosal function/electrophysiology were assessed over 24 h at 4 °C. ATP was consistently greater in Groups 2–4 than in the Control Group. Groups 3 and 4 exhibited significantly greater ATP, ATP/ADP ratios and energy charge levels after 12-h storage than in the other Groups. Histologic injury was generally lower in the AA-treated tissues (Groups 3 and 4); after 24 h, only minor epithelial clefting (Park's median grade 2) was present in Group 4; and consistent transmural infarction (grade 8) was evident in Groups 1 and 2. Combined treatment with luminal amino acid solution and oxygenated storage solution (PFC or AA solution) significantly improves energetics and mucosal function. This strategy may have implications for successful SB preservation in the clinic.

10 Cancer Control spinal cord compression, and electrolyte abnormalities such as hypercalcemia or hypokalemia) to iatrogenic pharmacologic treatment) in nature.5 Patients with cancer, especially those with advanced cancer, often have multiple factors at play, such as opioid analgesic use, reduced food and fluid intake, reduced mobility, advanced age, or malignancy-related conditions (eg, partial bowel obstruction, tumor-related hypercalcemia, and chemotherapyinduced constipation).6 Constipated cancer patients may have poor performance status, which implies decreased mobility,as well as poor nutritional status,and they may be taking medications that contribute to constipation. Besides opioids, which comprise the most common class to induce constipation, many drug classes may result in constipation including chemotherapy agents, anticholinergics (tricyclic antidepressants,phenothiazines), calcium or aluminum-containing antacids, iron preparations, and antiemetics (5-HT3 antagonists) (Table 1). Constipation is probably the most prevalent side effect of opioid use, and it is also the side effect that is least likely for tolerance to develop.3 Thus, prophylactic laxatives should be used when administering such agents on a regular basis.2 Introduction

An amino acid-based solution has been recently developed and has demonstrated significant protective effects during cold storage of small bowel (SB). This study was designed to examine the role of this novel solution in ameliorating intestinal injury in an in vivo model of ischemia-reperfusion (IR). The impact of luminal treatment with an amino acid-based (AA) solution was assessed throughout reperfusion after 60-min warm ischemia (WI) in rodent SB. Energetics (ATP and total adenylates) remained significantly elevated throughout 60-min reperfusion in AA-treated tissue compared with untreated controls. Increases in end-products (ammonia and alanine) and increases in alanine aminotransferase and glutaminase activity implicated greater amino acid metabolism in AA-treated tissues. After reperfusion, malondialdehyde levels were similar between all groups. Glutathione levels were consistently elevated in AA-treated tissues and by 60 min reperfusion values were sixfold greater than control. AA-mediated protection during IR resulted in reduced neutrophil infiltration suggesting a weaker inflammatory response. Barrier function and electrophysiology parameters exhibited a clear pattern of mucosal preservation in AA-treated tissues; histology supported these findings. This study raises the possibility of a role for a luminal nutrient-rich solution during ischemic storage of small bowel in the clinic.

During procurement, isolation, and transplantation, islets are exposed to high levels of oxidative stress triggering a variety of signaling pathways that can ultimately lead to cell death. Glutamine is an important cellular fuel and an essential precursor for the antioxidant glutathione. The aim of this study was to examine the role of intraductal glutamine administration in facilitating recovery of isolated rat islets from pancreases subjected to a clinically relevant period of warm ischemia. Islets were isolated in Sprague-Dawley (SD) rats (n= 18 per group). Pancreata in groups 1 and 2 were procured immediately while groups 3 and 4 were subjected to 30-min warm ischemia. Groups 2 and 4 were treated intraductally with 5 mM glutamine prior to pancreatectomy. Exposure to 30-min warm ischemia significantly reduced islet yield [groups 1 & 2 (nonischemia): 503 ± 29 islets/rat vs. groups 3 & 4 (ischemia): 247 ± 26 islets/rat; p < 0.05]. Intraductal glutamine treatment significantly improved islet yield when pancreata were subjected to 30-min warm ischemia [144 ± 16 islets/rat without glutamine (group 3) vs. 343 ± 36 islets/rat with glutamine (group 4), p < 0.05]. Glutamine also significantly improved islet viability (values were 50 ± 4% in group 4 vs. 27 ± 3% in group 3, p < 0.05). Similarly, glutathione (reduced) levels were significantly elevated in both glutamine-treated groups; however, this increase was greatest in tissues exposed to ischemia (2.76 ± 0.04 nmol/mg protein in group 4 vs. 1.66 ± 0.04 nmol/mg protein in group 3, p < 0.05). Intraductal glutamine administration considerably improves the islet yield, viability, and augments endogenous glutathione levels in pancreata procured after a clinically relevant period of ischemia. Intraductal administration of glutamine at the time of digestive enzyme delivery into the harvested pancreas may represent a simple yet effective tool to improve islet yields in clinical isolations.

A two-layer cold storage method (TLM) allows sufficient oxygen delivery to pancreata during preservation and resuscitates the viability of ischemically damaged pancreata in the canine pancreas transplant model. In this study, we applied a short-term preservation of the TLM to human pancreata after prolonged cold ischemia prior to islet isolation, and investigated the mechanisms of resuscitation of the ischemically damaged human pancreas by the TLM. Human pancreata were procured from cadaveric donors and preserved by the TLM for 3.2 ± 0.5 h after 11.1 ± 0.9 h of cold storage in UW (TLM group), or by cold UW alone for 11.0 ± 0.3 h (UW group). Islet isolations of all pancreata were performed using the Edmonton protocol. Islet recovery and in vitro functional viability of isolated islets were significantly increased in the TLM group compared with the UW group. According to the criteria of the Edmonton protocol, 10/14 cases (71%) in the TLM group were transplanted to patients with type I diabetes mellitus compared with only 5/21 cases (24%) in the UW group. In the metabolic assessment of human pancreata, levels of energetic parameters (ATP, total adenylates, and energy charge) were significantly increased, and malondialdehyde (MDA) levels were significantly decreased after the TLM preservation. There was no observable change in the incidence or degree of mitochondrial injury after the TLM preservation. Additional short-term storage by the TLM resuscitates the ischemically damaged human pancreas by regenerating the energetic status and prevents further damage by oxidative stress, ultimately leading to improvements of islet recovery and in vitro function. Use of the TLM following prolonged storage in UW provides an excellent adjunctive protocol for treating human pancreata for the rigors of the islet isolation process.

The prevalence of patients with cancer histories and types of cancers prevailing among a cohort of adults with end-stage hip osteoarthritis was established in order to determine if this group might require some form of enriched pre- and postoperative rehabilitation in view of their adverse medical history. Body weights and selected physical capacity indicators were specifically compared among hip surgical patients with and without cancer histories to specify characteristics that could direct potentially desirable and improved intervention efforts. The medical records of 1,000 hip osteoarthritis surgical candidates were scrutinized, and numbers with and without malignancy histories were recorded. Malignancy typologies and selected body mass and physical capacity indices were recorded. Specific subgroup comparisons among these variables were then made for 40 cancer survivors and an age- and gender-matched subgroup of 40 otherwise healthy osteoarthritis patients, and for selected breast, prostate, and colon cancer survivors. (1) Fourteen percent of the present patient group had a cancer history. (2) The most common malignancy noted was breast cancer, followed by prostate and then colon cancer. (3) Among subjects matched for age and gender, 85% with a cancer history were overweight or obese, compared with 60% of those with no comorbid disease history. (4) Patients with cancer histories were more impaired immediately before, and after, surgery than patients with no cancer history. (5) Patients with breast and colon cancer histories had significantly slower recovery rates after hip surgery than those with a prostate cancer history (p0.05). Thus, breast, prostate, and colon cancer survivors constitute a modest proportion of patients undergoing surgery for painful disabling hip osteoarthritis. As a subgroup, cancer survivors, especially breast cancer survivors, are overweight, and more impaired before and after surgery than adults of the same age without a cancer history undergoing hip surgery.

Aurelia aurita is a scyphozoan, abundant in the Mexican Caribbean during summer. Although usually innocuous, there is evidence of it causing harm to humans. This work investigates the biological activities of crude and fractionated extracts of A. aurita. Live specimens were collected between July and September 1999 from the Mexican Caribbean. The tentacular margin was dissected immediately and frozen at -50oC. A nematocyst suspension was prepared, discharged, and the supernatants lyophilized. Hemolytic assay was performed with lyophilized crude extract on bovine, sheep, and human red blood cells. Erythrocyte sensitivity to the toxin was ranked in descending order: human, sheep, and bovine. Toxic activity on Artemia nauplii was evaluated using the same crude extract for different exposure periods (3, 5, and 10 hours); only 48 and 72 hour old Artemia nauplii showed 50% mortality. Partial toxin purification was completed by sequential liquid chromatography using three gels (Sephadex G-200, DEAE Sephadex A-50, and Sephadex G-100). Intramuscular neuroactivity was detected in the crab Ocypode quadrata for two partially purified fractions. These fractions were found to have molecular weight components of 66 and 45 kDa, respectively.

Purpose: To perform a dosimetric and radiobiological comparison of the composite prostate plans when the boost plan was optimized with and without knowledge of the initial dose distribution. Methods: Ten patients previously treated in our department were selected for this study. The planning target volume (PTV) included the prostate, seminal vesicles and pelvic lymph nodes (PTV1) while the prostate with margins was considered PTV2. All organs at risk (OAR) and target structures were contoured by the prescribing physician. The prescribed doses were 54Gy in 30 fractions to PTV1 and 78Gy to PTV2. All plans were optimized using the RayStation treatment planning system (TPS). Plans to deliver the prescribed dose to PTV1 were optimized first. Two boost plans and the respective composite plans were then created for each patient. First, the boost plan was optimized by treating the prescription of 24Gy to PTV2 as a standalone plan. Second, the boost plan for PTV2 was optimized taking into consideration the dose distribution of the initial plan. For each method the respective composite plans was created. The two final composite plans were compared by in terms of DVHs and isodose line distributions, as well as radiobiologically. Results: In terms of PTV2 coverage the two methods were very similar with negligible differences. However, differences were observed for the overall coverage of PTV1, seminal vesicles and penile bulb. For these structures the mean doses were higher in the case of the independent boost plan. Similar trend was observed for the doses to bladder and rectum but the differences were smaller. The dosimetric differences were patient (OAR) dependent. Conclusion: By including the initial dose distribution during boost plan optimization the final composite plan shoed that doses to most OAR can be reduced. The dosimetric differences are patient dependent and in some cases can be significant.

Background Targeting interleukin-6 receptor signal transduction by tocilizumab (TCZ) has been shown to be an effective treatment for patients with rheumatoid arthritis (RA). Objectives This study aimed to describe safety and efficacy of tocilizumab (TCZ) in patients with rheumatoid arthritis (RA) in routine clinical practice, based on prospectively registered observational data from a third-level hospital. Methods Forty patients with RA who were started on TCZ at the Rheumatology Unit between April 2009 and July 2011 were included. Changes in tender joint count (TJC), swollen joint count (SJC), erythrocyte sedimentation rate (ESR), haemoglobin (Hb), 28-joint Disease Activity Score (DAS28), European League Against Rheumatism (EULAR) response and remission rates with the 2011 American College of Rheumatology (ACR)/EULAR criteria after 4, 12, 24, 48, 72 and 96 weeks (W) were investigated. SPSS 17.0 program was used for statistical analysis. Adverse Events (AE) and withdrawals were also described. Results Thirty-five women and five men were included. Median age was 52 years old (33-79). Median disease duration was 11 (2-39) years. Rheumatoid Factor was positive in 32 patients (80%), anti-citrullinated protein antibody in 34 (85%) and anti-nuclear antibodies were detected in 34 patients (85%). Thirty-four patients (85%) had previously received at least one biological treatment. Decrease of TJC, SJC, ESR and DAS28 was statistically significant in all the weeks. Increase of Hb was statistically significant until W48. By analysing the response data the following results were obtained: Regarding the use of concomitant treatments, the following information compares the first and the last data recorded in each patient: disease-modifying antirheumatic drugs in 60% to 48%; nonsteroidal anti-inflammatory drugs in 80% to 35%; the average dose of corticosteroids (adjusted to prednisone dose) was initially 6.1 mg/day and finally 4.2 mg/day. Regarding safety, 31 patients (77.5%) had at least one AE. The most frequent AE were infections (22 episodes), dyslipidaemia (present in 16 patients) and haematology abnormalities (detected in 14 patients). Three severe AEs were observed (community-acquired pneumonia, fever without source and acute toxic hepatitis). There was no mortality in the group. Fifteen patients (31%) withdrew from treatment for safety-related (25%) or non-safety-related (12.5%) reasons. Conclusions In our serial TCZ had achieved a good-or-moderate EULAR response in patients with RA in routine clinical practice. An early and maintained response was observed. The most frequent AE were infections, dyslipidemia and haematological abnormalities. The main causes of withdrawal were AEs. Disclosure of Interest None Declared