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Oxidation of LDL in the arterial extracellular matrix is a key event in the development of atherosclerosis and autoantibodies against oxidised LDL antigens reflect disease severity and the risk of developing acute cardiovascular events. Since type 2 diabetes is associated with increased oxidative stress, we tested the hypothesis that autoantibodies against oxidised LDL antigens are biomarkers for vascular complications in diabetes.We studied 497 patients with type 2 diabetes without clinical signs of coronary heart disease. Oxidised LDL autoantibodies were determined by ELISA detecting IgG and IgM specific for native and malondialdehyde (MDA)-modified apolipoprotein B-100 peptides p45 and p210. The severity of coronary disease was assessed as the coronary artery calcium score.Patients affected by retinopathy had significantly higher levels of IgG against MDA-p45 and MDA-p210. In contrast, high levels of autoantibodies against the corresponding native peptides were associated with less coronary calcification and a lower risk of progression of coronary disease.Our observations suggest that LDL oxidation is involved in the pathogenesis of diabetic retinopathy and that autoantibodies against apolipoprotein B peptides may act as biomarkers for both micro- and macrovascular complications in diabetes.

Introdução: A Anemia Aplásica (AA) é uma doença rara, sem predominância degênero, com maior incidência na infância e após os 60 anos, especialmente napopulação asiática. A AA pode ser classificada pelo grau de acometimento(Moderada, Severa, Muito Severa) e pela etiologia, afetando o prognóstico e aconduta terapêutica. A diferenciação é baseada em hemogramas, testes genéticos eimunológicos, além dos diagnósticos diferenciais como citopenia refratária esíndromes mielodisplásicas hipoplásicas que também devem ser excluídos porexames clínicos e laboratoriais. O Transplante de Células-Tronco Hematopoiéticas (TCHT) é um tratamento eficaz, especialmente em crianças, com taxas de sucessode 90% pela literatura internacional quando há doador aparentado idêntico.Transplantes haploidênticos ou de doador Não Aparentado (NAP) são consideradosem casos mais graves. No Brasil, as taxas de eficácia e sobrevida do TCHT pediátrico para AA são inferiores às observadas nos Estados Unidos e na Europa. Objetivos: Avaliar a sobrevida de pacientes pediátricos com AA após TCHT,analisando fatores como a origem do enxerto e a compatibilidade do doador. Materiais e métodos: Estudo realizado em um centro de saúde terciário, de janeirode 2021 a junho de 2024. Incluiu pacientes com AA até 21 anos submetidos aTCHT. Pacientes com outras doenças hematológicas, maiores de 21 anos ou quenão passaram por TCHT foram excluídos. Analisou-se a sobrevida, a origem doenxerto e a compatibilidade do doador, com metodologia estatística descritiva.Resultados: Foram identificados 11 pacientes com AA submetidos a TCHT, comquatro apresentando falha de pega, resultando em 15 TCHT no total. Um foiautólogo e 14 alogênicos (7 haploidênticos, 4 NAP, 2 aparentados). 9 enxertosforam de medula óssea e 6 de sangue periférico. 3 pacientes faleceram (28%), doisapós o segundo TCHT, com todos os óbitos relacionados ao transplante erealizados com sangue periférico e doadores haploidênticos. Discussão: O estudoconfirma a eficácia do TCHT no tratamento de AA em pediatria e a maiormortalidade desse tratamento na população brasileira. A escolha da fonte decélulas-tronco e a compatibilidade do doador são críticas para a sobrevida. Melhores resultados foram observados com medula óssea em comparação comsangue periférico, alinhando-se com a literatura. A falha de pega em quatropacientes e os óbitos sugerem necessidade de seleção cuidadosa de doadores emanejo pós-transplante rigoroso. Os óbitos podem apontar que transplantes comsangue periférico podem apresentar maior risco; entretanto, essa observação podeser uma limitação do estudo devido à pequena amostragem, já que todos os óbitosocorreram após o segundo TCHT, os quais todos foram realizados com sangueperiférico. Portanto, há necessidade de mais estudos para entender melhor osfatores que contribuem para a maior taxa de óbitos entre pacientes brasileirossubmetidos ao TCHT para tratamento de AA. Conclusão: A escolha da origem doenxerto e da compatibilidade do doador é fundamental para o sucesso do TCHT empacientes pediátricos com AA. Enxertos de medula óssea e doadores com maiorcompatibilidade resultam em melhores desfechos. As taxas de mortalidade sugerema necessidade de estratégias de tratamento individualizadas e gestão rigorosa dosriscos. Futuras pesquisas devem focar na otimização do tratamento para melhorar asobrevida e a qualidade de vida dos pacientes brasileiros.

Introduction: Allogeneic Stem Cell Transplant (ASCT) is a curative strategy for varioushematological diseases. Following ASCT, chimerism analysis becomes instrumental inassessing engraftment success, guiding immunosuppression strategies, anddetermining the necessity of interventions such as Donor Lymphocyte Infusion (DLI). Inleukemia cases, chimerism evaluation is a crucial tool for predicting and monitoringdisease relapse. However, the routine chimerism and its efficacy in the pediatricpopulation remain subjects of ongoing debate, necessitating further studies. Objective: Assess the correlation between chimerism levels, timing of testing, and their predictivevalue for disease relapse and survival among pediatric patients with leukemiapost-ASCT. Materials and methods: This retrospective study enrolled pediatric patientsdiagnosed with leukemia who underwent ASCT. Patients with other hematologicaldiseases and those who received autologous stem cell transplants were excluded fromthe analysis. This retrospective study was conducted in a tertiary care center, and thedata was collected from 2021 until January 2024. Demographic characteristics, survival,and clinical/laboratory findings, including chimerism analysis of 1-year after ASCT viaimmunophenotyping using flow cytometry. The Chimerism analysis was performed at sixcheckpoints (D+30, D+60, D+100, six months, nine months, and 12 months). Thestatistical analysis was made using descriptive statistics. Results: Of the 90 patients cohort, 85 (94%) patients underwent a single transplant, while 5 (6%) patients had twobone marrow transplants. Among the patients who achieved successful engraftment, 19 (21%) experienced disease relapse, with 9 relapses occurring within the first yearpost-ASCT. Notably, all relapses between D+100 and 12-months post transplantoccurred in patients with complete chimerism. The 9-month checkpoint exhibited thelowest patient attendance, with only 44% of patients having complete chimerism dataand 36 missing results. Conversely, the D+60 checkpoint had the fewest missing tests,with only 13 absences. The initial three checkpoints (D+30, D+60, D+100) demonstrated higher attendance rates than later. The majority of deaths occurred duringthe D+100 period, with 6 out of 9 relapses occurring during this timeframe. It was alsothe period with the higher percentage of mixed chimerism, having 22% in the D+30, 21% in the D+60, and 17% in the D+100. Conclusion: Overall, these findings suggestthat chimerism analysis, particularly at critical time points such as D+100, is valuable tohelp predict disease relapse in pediatric patients post-transplant and their survival.However, challenges in patient attendance at certain checkpoints indicate the need forimproved monitoring strategies to optimize patient outcomes.

Objetivos: A leucemia mieloide aguda (LMA) incide sobre a faixa etária pediátrica com elevada morbimortalidade e taxas de cura subotimas. O protocolo do Grupo de Estudos de Leucemia Mieloide Aguda Infantil (GELMAI) propõe a redução da toxicidade do tratamento e de seus desfechos adversos durante a fase de indução. Este trabalho visa à análise dos efeitos colaterais apresentados pelos pacientes submetidos ao protocolo GELMAI, com ênfase na incidência de infecções e na toxicidade hepática, cardíaca e hematológica. Métodos: Trata-se de um estudo transversal do tipo série de casos dos pacientes menores de 18 anos, portadores de LMA, com diagnóstico confirmado entre julho de 2021 e julho de 2023, em tratamento pelo protocolo GELMAI no HUOC. Excluídos os pacientes com LMA associada a Síndrome de Down e com leucemia promielocítica. Resultados: Foram incluídos no estudo 7 pacientes, com idade média de 14 anos ao diagnóstico. Foi evidenciada leucocitose em 57% dos pacientes, e o mielograma demonstrou em média 53% de infiltração blástica. O subtipo FAB mais prevalente foi o M2 (4/7), destes, t(8;21) estava presente em todos. Foram reportadas demais alterações citogenéticas: o cariótipo complexo, o rearranjo MLL e a trissomia do 8. A sobrevida global correspondeu a 5/7. Em relação às infecções, mais incidentes entre as 2ª e 3ªintensificações. As septicemias representam 43,7% do total de eventos infecciosos, bem como a causa de óbito de 1 paciente da amostra, cujo agente etiológico não foi definido. As infecções bacterianas assumiram maior prevalência (40% dos casos), comparando-se às infecções fúngicas (13,3%). Na amostra estudada, 1 paciente apresentou insuficiência cardíaca no momento do diagnóstico, apresentando Troponina I com valor de 1207,9 pg/mL; fração de ejeção de 38%, disfunção sistólica e diastólica moderadas de ventrículo esquerdo. Este paciente evoluiu com melhora clínica durante dois ciclos de indução do GELMAI. Quanto à hepatotoxicidade, ocorreu em 1 paciente, com hiperbilirrubinemia total acima de 10 vezes o valor de referência do serviço (1,2 mg/dL). Esse evento ocorreu durante a fase da 2ªintensificação, em uso de Citarabina e Etoposide, drogas com potencial hepatotóxico e, foi descartada etiologia viral. Discussão: Observou-se predominância de pacientes adolescentes, o que coincide com o segundo pico de incidência da LMA, atrelado a prognóstico mais reservado, devido a maiores índices de toxicidade ao tratamento. Os parâmetros citogenéticos e moleculares encontrados representaram amostra predominante de prognóstico inicial favorável. A prevalência do subtipo FAB-M2, que tem diferenciação maior em relação aos demais, justifica a média inferior ao descrito na literatura de infiltração blástica ao mielograma. Os eventos infecciosos e as toxicidades encontradas ocorreram dentro do previsto. Não houve mortalidade durante a etapa de indução, a qual atingia 45% com demais protocolos aplicados no HUOC. Conclusão: Desfechos adversos e toxicidades constituem preditores de menores taxas de sobrevida e de mortalidade precoce, sobretudo nos países em desenvolvimento. Desta maneira, o estudo visa colaborar à elaboração de uma rede de vigilância que previna as principais complicações advindas do tratamento da LMA pelo GELMAI, através da identificação dos fatores de risco e complicações.

Introdução: O monitoramento da Doença Residual Mensurável (DRM) tornou-se prática clínica de rotina no tratamento das Leucemias Linfoblásticas Agudas (LLA). A avaliação da DRM tem demostrado fator prognóstico, que permite a atribuição de grupos de risco nos diferentes braços de tratamento, variando desde redução significativa do tratamento a intensificação. Além do uso em pacientes com LLA recaída e pacientes submetidos à transplante de células-tronco, e mais recentemente nos casos de tratamento com células CAR-T. A imunofenotipagem por Citometria de Fluxo Multiparamétrica (CFM) tornou-se uma ferramenta importante no monitoramento na pesquisa da DRM (Doença Residual Mensurável), no monitoramento das células imunes infundidas (CAR) e na avaliação da resposta imunológica pós CAR-T. Relato de dois casos: Paciente 1, masculino, 16 anos. Diagnóstico em 2017 de LLA B. Transplante de medula em 09/2021. Apresentou recaída combinada (medula e sistema nervoso central) pós TMO haploidêntico em 10/2022. DRM pré infusão era de 0,38%. Terapia CAR-T em 17/01/2023. Condicionamento: Fludarabina (FLU) 120 mg/m2 e Ciclofosfamida (CY) 1.000 m2. Intercorrências durante o internamento: febre diariamente de 20/01 a 24/01 e Hipotensão (83/35), resolvida com uma fase rápida. Grau I na classificação de CRS (síndrome de liberação de citocinas). Não apresentou sinais de neurotoxicidade. Alta no D+10. Necessitou de duas transfusões de plaquetas e uma infusão de imunoglobulina. DRM 30 dias pós CART – negativa. Segue em acompanhamento ambulatorial com DRM negativa. Paciente 2. LLA-B BCR:ABL positivo. Em 2018 troca Imatinibe por Desatinibe devido toxicidade hematológica. E no mesmo ano recaída em SNC e MO. TMO Haploidêntico em 01/2019. Recaída em SNC em 2022. Tratamento com MADIT e radioterapia (24 Gy em 12 frações). DRM pré infusão era negativa. Realizou terapia CAR-T em 02/2023. Condicionamento: Flu 120 mg/m2 e Cy 1.000 m2. Não teve intercorrências durante o internamento (10 dias). Transfusões: zero. Não apresentou sinais de CRS e neurotoxicidade. Citometria de Fluxo: Foi realizada a técnica de lise total e painel de anticorpos monoclonais padrão Euroflow. Citômetro de Fluxo FACS Canto II (BD). Os anticorpos monoclonais utilizados: Painel de anticorpos monoclonais utilizado: CD3, CD10, CD19, CD20, CD22, CD24, CD34, CD38, CD45, CD66c/CD123, CD73/CD304, CD81, CD79a (citoplasma). As DRMs foram negativas nas amostras analisadas nos dias D+30 e D+60 e D+90. Conclusão: O acompanhamento clínico ao diagnóstico e durante a avaliação contínua nas diferentes fases da doença são fundamentais para o êxito no desfecho. A Citometria de Fluxo Multiparamétrica (CFM) é uma ferramenta importante não só no diagnóstico da LLA mas também na avaliação da DRM, através de técnicas e painéis padronizados e o acompanhamento da DRM tem demonstrado a eficácia do tratamento.

Objective: This study compares the dose intensity of induction regimens by evaluating outcomes such as early deaths, death in the first remission, and survival. The results of this study will contribute to the development of therapeutic protocols with a reasonable balance in the intensity of induction, allowing remission to be achieved with less toxicity, adapting and establishing the treatment for acute childhood myeloid leukemia in developing countries. Methods: Pubmed and Scopus were used for systematic searches performed in January 2021. Descriptors involving specific or synonymic words such as Acute myeloid leukemia, pediatric, induction, and dose-intensity combined with Boolean operators AND and OR were used, without limits of time-frame or language. All relevant studies were read in full, and the data extraction was performed for those that fulfilled the inclusion criteria (patients aged between 0–21 years old, diagnosed with AML, describing results of induction regimens where one of the groups – intervention or comparison –included different dosing for the same chemotherapy drug). Were excluded the studies publishing exclusively Down's Syndrome patients and acute promyelocytic leukemia diagnosis, single-arm studies (with no comparator group), and studies where the intervention group had a different chemotherapy regimen than the comparator group. Results: The initial search retrieved 1185 papers. In the second round of screening of the remaining 207 articles, full-text papers were assessed for eligibility. From the nine papers included with dose modification during the induction phase of chemotherapy treatment, six referred to changes on cytarabine (ARA-C), two on idarubicin (IDR), and two on daunorubicin (DNR) (one of these studies simultaneously changed ARA-C and DNR). Results showed no significant difference between standard vs. high doses for CR and death during induction. Relapse rates were higher in the standard-dose than in the high-dose arms (1.44; 95% IC: 1.22–1.69). Despite the higher risk of relapse in the standard-dose group, event-free survival analysis shows no significant difference between the arms of each study. Discussion: The outcome of AML treatment in children and adolescents has improved over the last decades. Overall survival rates are now superior to 70% due to implementing strategies such as intensification of therapy, refinements in supportive care, and more precise risk classification. However, treatment-related deaths are exceptionally high in places with limited resources. Even regimens with standard-dose chemotherapy are associated with life-threatening complications due to profound myelosuppression and organ dysfunction. Establishing the best combination and sequence of treatment components, including hematopoietic stem cell transplantation, remains an unmet need. Conclusion: There is no significant difference between standard vs. high doses of chemotherapy during induction for selected outcomes such as complete remission and event-free survival. Reducing the dose intensity of standard AML remission induction regimens should be investigated in regions with limited resources.

A relationship was developed which accurately calculates the effective dielectric constant ℇeff as a function of width-to-height (b/t) ratios for printed structures such as bow-tie antennas on electrically thin substrates. The impedance and impedance–frequency relation of bow-tie antennas on electrically thin substrates were experimentally determined. Radiation patterns of bow-tie antennas and collinear bow-tie antenna arrays on electrically thin substrates were also measured, and the results are presented. © 1998 John Wiley & Sons, Inc. Microwave Opt Technol Lett 17: 359–368, 1998. [ABSTRACT FROM AUTHOR]

El Tor strains of Vibrio cholerae are capable of producing a haemolysin which they actively secrete into the growth medium. This requires translation to produce the protein at the surface of the cytoplasmic membrane and translocation across this membrane, the periplasmic space and the outer membrane. The mechanism by which this occurs is poorly understood. In addition to the structural gene for the haemolysin (hlyA), we have cloned a second adjacent gene, hlyB. By site-directed mutagenesis, specific hlyB mutants have been constructed. These mutants are defective in the secretion of HlyA in the early to mid-exponential phase of growth and the haemolysin becomes trapped within the cell and is only released in stationary phase. Nucleotide sequence analysis and cell fractionations reveal HlyB to be a 60.3 kD putative outer membrane-associated protein. [ABSTRACT FROM AUTHOR]

The El Tor haemolysin, product of hlyA, is exported from Vibrio cholerae as a Mr 80000 protein which can be subsequently cleaved to give two proteins of Mr 65000 and 15000. Nucleotide sequence analysis has demonstrated that hlyA encodes a protein of Mr 82250 with a potential 18-amino-acid signal sequence. The non-haemolytic classical strain 569B has been shown to have a structural gene defect rather than a defect in secretion. By non-reciprocal recombination it was possible to transfer this defect onto a plasmid and show that a truncated hlyA product of Mr 27000 is made in Escherichia coli K-12 minicells. Nucleotide sequence analysis demonstrates an 11-base-pair deletion which would result in a Mr 26940 protein probably loosely associated with the membrane. [ABSTRACT FROM AUTHOR]

The present study aimed at assessing the effects of methionine and arginine on the immune response of broiler chickens submitted to immunological stimuli. Three methionine concentrations (0.31, 0.51, and 0.66% from 1 to 21 days of age; 0.29, 0.49, and 0.64% from 22 to 42 days of age) and 2 arginine concentrations (1.33 and 1.83%; 1.14 and 1.64% for the same life periods) were tested. Birds were divided into two groups for immunological stimuli (3x2x2 arrangement). Vaccines against Marek's disease, fowl pox, infectious bronchitis, Freund's Complete Adjuvant, Sheep red blood cells (SRBC), and avian tuberculin were administered to one group as immunological stimuli; the other group did not receive any stimulus. The experiment was carried out with 432 one-day-old male Ross broilers, distributed into 12 treatments with 6 replicates of 6 birds each. Performance data were weekly collected. Anti-SRBC antibodies were collected by hemagglutination test and cell immune response (CIR) was measured by tubercularization reaction in one wattle 24 hours after administration of the second tuberculin injection at 42 days of age. The weight difference between the two wattles of each bird (one injected with tuberculin and the other not) was the measure of CIR. Arginine levels did not influence either bird performance or immune response. Methionine concentrations higher or lower than usually adopted in broiler production (0.51 and 0.49%) equally failed to influence the birds' immune humoral response, but the best CIR was observed at the intermediate methionine level. Vaccines administered on the first day of age impaired bird performance up to the 21st day of age.

Abstract Background Endothelium dysfunction is believed to play a role in the development of cardiovascular disease. The aim of the present study was to evaluate the suitability of organ culture as a model for endothelium dysfunction. Methods The isometric tension was recorded in isolated segments of the rat mesenteric artery branch, before and after organ culture for 20 h. Vasodilatation was expressed as % of preconstriction with U46619. The acetylcholine (ACh) induced nitric oxide (NO) mediated dilatation was studied in the presence of 10 μM indomethacin, 50 nM charybdotoxin and 1 μM apamin. Endothelium-derived hyperpolarising factor (EDHF) was studied in the presence of 0.1 mM L-NOARG and indomethacin. Prostaglandins were studied in the presence of L-NOARG, charybdotoxin and apamin. Results The ACh-induced NO and prostaglandin-mediated dilatations decreased significantly during organ culture (NO: 84% in control and 36% in cultured; prostaglandins: 48% in control and 16% in cultured). Notably, the total ACh-dilatation was not changed. This might be explained by the finding that EDHF alone stimulated a full dilatation even after organ culture (83% in control and 80% in cultured). EDHF may thereby compensate for the loss in NO and prostaglandin-mediated dilatation. Dilatations induced by forskolin or sodium nitroprusside did not change after organ culture, indicating intact smooth muscle cell function. Conclusions Organ culture induces a loss in NO and prostaglandin-mediated dilatation, which is compensated for by EDHF. This shift in mediator profile resembles that in endothelium dysfunction. Organ culture provides an easily accessible model where the molecular changes that take place, when endothelium dysfunction is developed, can be examined over time.

Background: Care transitions are a challenging and crucial point for many high-risk patients; errors in medication use can result in preventable hospital readmissions, which worsen patient outcomes and result in decreased reimbursement and increased expenses for health systems. Transitions of care (TOC) is an opportunity where pharmacists in the outpatient setting can prevent medication errors and decrease hospital readmissions., Objective: The primary objective of this study was to evaluate the impact of pharmacist-conducted comprehensive medication reviews (CMRs) on 30-day hospital readmission rates. Secondary objectives included medication therapy problems (MTPs) identified, recommendations or interventions made, and impact on cost savings., Methods: Patients discharged from an Intermountain Health hospital with a high readmission risk score, value-based insurance plan, and primary care provider (PCP) at one of 21 primary care clinics were identified using an internal report. Patients were contacted by a pharmacist after discharge for a CMR; pharmacists then relayed any MTPs and therapeutic recommendations to the PCP before the patient's follow-up appointment. Data were retrospectively collected and analyzed., Results: A total of 2717 discharges occurred throughout Intermountain Health and affiliate clinics from October 5, 2020, to January 31, 2021; 30-day readmissions rates in the pharmacist intervention group versus the comparator group were 24/191 (12.57%) versus 511/2526, (20.23%), respectively, yielding a statistically significant difference (P < 0.05). The absolute risk of readmission was reduced by 7.66%, with a number needed to treat of 14. Approximately 14 readmissions were prevented in the pharmacist intervention group, resulting in an estimated cost savings of $212,800 and return on investment of $16.19:1., Conclusion: Pharmacist intervention in the outpatient setting in high-risk TOC patients resulted in decreased 30-day readmission rates and increased cost savings. Further investigation is warranted to identify patient factors where pharmacist intervention is most beneficial., (Copyright © 2023 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.)

Background: A common neurological condition worldwide is Reward Deficiency Syndrome (RDS) leading to both substance and non-substance addictive behaviors, that must be combatted by integrating both central nervous system and peripheral nervous system biological approaches. Integrity of hemoglobin is a crucial determining factor for the overall health functions. Nutrient repletion therapy should be a fundamental strategy to restore the healthy properties of blood. A unique patent-pending iron-free VMP35 formulation was engineered by our laboratory to restore iron-dependent hemoglobin in anemic cells using a proprietary Prodosome ® absorption technology. This formulation, containing an array of nano-emulsified botanical ingredients rich in bioflavonoids, strengthens the structural integrity of connective tissues, and potentiates immune competence, cellular aerobic metabolism, and enhances efficient regulation of inflammatory events. We discuss the intricate aspects of strong vs. fragile immunity and consequential inflammatory responses to convey a deeper understanding of the varied and overly complex sequela of immunological behaviors and events. The effect of the VMP35 is mediated through highly absorbable nutritional/nutrigenomic repletion enabling improvements in the systemic set of functional behaviors. In fact, the iron-free VMP35 facilitates a "Systems Biology Approach" which restores hemoglobin status, reverses anaerobic hypoxia, improves competent immune responsivity, and regulates appropriate and controlled activation of general and neuro-inflammatory sequela. Under these pathogenic circumstances, iron-deficiency anemia has been misconceptualized, and a new nosological term, Chronic Anemia Syndrome, is proposed. The comparative therapeutic rationale of Reductionist vs. Systems Biology approaches is also explained in detail., Methods: The efficacy of the novel therapeutic iron-free VMP35 liquid nutraceutical is detailed in restoring iron-dependent hemoglobin to RBCs and boosting cellular morphology, viability, and immune competence, thereby reducing the need for prolonging inflammatory sequela., Results: This was demonstrated in a previous IRB approved multi-subject human study. In addition, two recent case studies report dramatic restorative benefits of nutrient repletion therapy of the VMP35 on subjects having experienced near-fatal events, which confirmed the findings explained in this manuscript., Conclusions: This novel iron-free VMP35 modulates an array of homeostatic biological parameters such as enhanced hemoglobinization, aerobic metabolism, viral immuno-competence, and inflammatory regulation. Further research, examining mechanistic and beneficial effects in athletic performance, is in progress. Importantly, during these troubled immune challenging times, modulating an array of homeostatic immunological and inflammatory dysfunctions are tantamount to improved population outcomes., Trial Registration: The Clinical investigation in a total of 38 subjects was conducted under an Institutional Review Board (IRB) from the Path Foundation in New York, NY (#13-009 April 25, 2013). The two case studies were done at Lancaster General Hospital, Lancaster, PA, and Jefferson University Hospital, Philadelphia, PA, USA. Both studies were retrospectively registered., Competing Interests: Competing interests Both Drs. Bruce S. Morrison, DO and Jeffrey Galvin, MD are highly respected board-certified practicing physicians whose clinical experience with the VMP35 confirmed the concept and hypothesis of this article and guided the writing of the manuscript. Bernard W. Downs is employed by Victory Nutrition International, Inc. (VNI), Lederach, PA, Debasis Bagchi, a professor in the Department of Pharmacological & Pharmaceutical Sciences, University of Houston college of Pharmacy and Steve Kushner of ALM R&D, are both scientific consultants to VNI, all of whom contributed to the concept of the manuscript respectively. Dr. Manashi Bagchi is an independent consultant, who compiled the data, wrote, and routed the manuscript to all concerned. Kourtney Randsdorp, and Justin Randsdorp are certified Occupational Therapists who were involved in the recovery treatment of DH in Case Study #1 and provided valuable suggestions and criticisms while drafting the manuscript.

Background: Enzymes are crucial for all aspects of metabolic function. Digestive enzymes from natural sources have been credited with beneficial effects in the digestion and absorption of food. N-SORB is a novel KD120 multienzyme complex (MEC) of metabolically activated enzymes composed of proteases, amylases, lipases, alpha-galactosidase, and glucoamylase from natural sources. These enzymes are encapsulated in a SK713 SLP (non-GMO soy lecithin phospholipid) absorption technology (Prodosome ® ). Objective: This randomized, double-blind placebo-controlled investigation assessed the safety and efficacy of N-SORB KD120 MEC in healthy male and female volunteers on various parameters of the blood, immunity, body composition, physical health, and quality of life following a 90-day intervention. Methods: Forty-six male and female (mean age: 25.8 ± 12.1 years) healthy volunteers were randomly assigned to receive either N-SORB (1 mL, twice daily) or placebo for 90 consecutive days. Complete blood count, as well as blood glucose, liver enzymes, and lipid profile were assessed pre- and post-intervention. Serum cytokine levels were determined by using a Bio-Plex Pro Human Cytokine 8-plex assay and enzyme linked immunosorbent assay (ELISA). Whole body composition analysis was performed by dual-energy x-ray absorptiometry (DEXA) to determine body fat mass, lean mass, and android and gynoid fat. Body weight, blood pressure, and physical health were assessed. Changes in quality of life were examined using the World Health Organization Quality of Life-abbreviated version and sleep quality was assessed using the 24-item Pittsburgh Sleep Quality Index (PSQI) questionnaire. Adverse events were monitored before, during, and after completion of the study. Results: Of the 46 subjects enrolled, a total of 40 subjects successfully completed the study. Compared to placebo, changes in blood cell counts including hematocrit, hemoglobin, mean corpuscular volume, platelets, and lymphocytes provide evidence of some improvement. Quality of life (QOL) parameters showed a small but significant improvement in the N-SORB group. A significant increase was observed in aspartate aminotransferase level in the placebo group at the end of 90 days of treatment; however, no increase was observed in the N-SORB group. No significant changes in blood urea nitrogen, serum creatinine, alkaline phosphatase, alanine aminotransferase, and lipid profile were observed between the placebo and treatment groups before and following intervention. No adverse effects were reported. Conclusions: This randomized, double blind, placebo-controlled clinical study demonstrates that short-term intervention with N-SORB improves the QOL and PSQI in healthy volunteers and did not significantly alter cardiometabolic parameters, lipid profile, or body composition.

Background: Hypercholesterolemic mice lacking factors required for activation of CD4 + T cells are characterized by reduced development of atherosclerosis. Consequently, it has been assumed that atherosclerosis involves loss of tolerance against modified self-antigens generated in response to hypercholesterolemia and that presentation of such antigens on major histocompatibility complex class II (MHCII) leads to activation of proatherogenic Th1 cells. In this study, we wanted to determine the role of antigen presentation on MHCII in atherosclerosis development., Methods: Apolipoprotein E (ApoE -/- ) mice deficient for MHCII (ApoE -/- MHCII -/- ) were used to study the role of MHCII in atherosclerosis development., Results: Compared with ApoE -/- mice, ApoE -/- MHCII -/- mice had reduced levels of CD4 + T cells, immunoglobulin G and M levels, and Th1 and Th2 cytokines in plasma. CD8 + T cells were increased and regulatory T cells were reduced both in spleen and in lesions of ApoE -/- MHCII -/- mice. Decreased plasma levels of inflammatory cytokines in ApoE -/- MHCII -/- mice indicated reduced systemic inflammation. Despite this, ApoE -/- MHCII -/- mice had significantly more atherosclerosis as assessed by en face Oil Red O staining of the aorta (4.7±2.9% versus 1.9±1.3%; P<0.01) and cross-sectional area of subvalvular lesions (7.7±2.2×10 5 µm 2 versus 4.6±2.8×10 5 µm 2 ; P<0.05). Cell transfer and blocking antibody studies suggested that loss of regulatory T cells is the most important cause of aggravated atherosclerosis in ApoE -/- MHCII -/- mice., Conclusions: Our observations demonstrate that antigen presentation on MHCII has important protective functions in atherosclerosis and that this is primarily the result of activation of regulatory T cells. These findings have implications for understanding the possible risks and benefits of immunosuppressive therapy in patients with cardiovascular disease.

Background: The American Society of Hematology reported that according to the National Heart, Lung, and Blood Institute (NHLBI) anemia is the most common blood disorder, which affects more than 3 million Americans, while the Global Burden of Disease 2016 (GBD 2016) reported that iron deficiency anemia (IDA) is the leading cause of anemia, which affects 1.93 billion people worldwide. Anemia is intricately linked to chronic inflammation, chronic kidney disease, gastrointestinal and gynecological malignancies, and autoimmune disorders. Hemorrhagic anemia results in substantial loss of blood, which causes significant alterations in all blood parameters, including reduced iron. The other type of anemia is chronic anemia syndrome (CAS), which is a constellation of disorders and chronic inflammatory events caused by an increasing anaerobic/acidic environment (promoting the growth of anaerobic organisms), inducing a defensive expenditure of alkalinizing buffers in hemoglobin (i.e. histidine), to prevent a dangerous lowering of blood pH. In this process, iron is cleaved from heme groups and transferred out of blood circulation into other organs, like the liver, appearing to be IDA, where excessive accumulation can lead to hemochromatosis, also known as 'iron overload anemia'., Design: A pilot clinical study was conducted in 38 subjects (men = 10; women = 28; age = 22-82 years) to evaluate the rate of absorption and effects on blood of VMP35 multi-nutrient complex (MNC), a non-iron containing liquid nutraceutical supplement. Subjects consumed either placebo or VMP35 (30 mL) over a period of 0, 5, or 30 min., Methods: Changes in peripheral blood smears from 38 subjects were observed using live blood cell imaging (LBCI) with phase contrast microscopy. Adverse events were rigorously monitored., Results: VMP35 caused positive changes in the blood, including morphological, hematological (including restoration of hemoglobin), and rheological changes following 5 min of administration, which were sustained for at least 30 min., Conclusion: Overall, the non-iron containing VMP35 can induce improvements in blood properties and potential benefits for subjects even with compromised digestive systems. No adverse events were reported. Further research studies are in progress to explore the mechanistic insight., Competing Interests: JRC is a highly respected board-certified practicing medical doctor whose clinical experience with the VMP35 confirmed the concept and hypothesis of this article and guided the writing of the manuscript. BWD, DB and SK contributed to the concept of the manuscript and are engaged with Victory Nutrition International, Inc., Lederach, PA, and ALM R&D, Oldsmar, FL, respectively. TA is an integral part of Veritas Health Inc., Woodbridge, ON, who organized the live cell imaging procedures. MB (Dr. Herbs LLC, Concord, CA) is an independent consultant, who compiled the data and routed the manuscript to all concerned.

Metropolitan Council Environmental Services has observed poorer than expected dewatering performance at its Empire Plant. This plant has both anaerobic digestion and enhanced biological phosphorus removal in its treatment train. A research program using pilot-scale anaerobic digesters investigated potential solutions to the plant's poor dewaterability. The dewaterability goal was to increase the cake solids from 12% total solids (TS) to 16% TS or higher. This research investigated 20 different reactor conditions including chemical, feed sludge, and digested sludge treatments. At the pilot scale, unaerated storage of waste activated sludge prior to thickening and addition of ferric chloride to digestion was found to achieve dewatered cake solids of nearly 17% TS with the added benefit of reducing polymer demand. Issues including the amount of chemical required and the resulting volatile solids destruction influence the viability of the process change, so a full-scale pilot and financial analysis is recommended before making permanent process changes.

Context: Large epidemiological studies often collect non-fasting samples, although the reliability of biomarkers may be uncertain., Objective: To explore the reliability and reproducibility of a single measurement of selected biomarkers in a sub-sample of the Malmö Diet and Cancer cohort., Methods: We estimated single- and average-measures intraclass correlation coefficients (ICC) for oxidized (ox)-LDL, interleukin (IL)-1β, IL-6, IL-8 and TNF-α., Results: Single-measures ICC in non-fasting samples of ox-LDL, IL-1β, IL-6, IL-8 and TNF-α were the following: 0.85, 0.71, 0.61, 0.78 and 0.66 for men, and 0.67, 0.81, 0.87, 0.69 and 0.81 for women. Biomarkers at non-fasting and fasting samples were highly correlated (all r > 0.80)., Conclusions: The observed ICC suggest that most of the examined biomarkers (non-fasting blood) would allow meaningful analysis in epidemiological studies.

Background and Purpose: Treatment with IgG against the malondialdehyde (MDA)-modified apolipoprotein B-100 epitope p45 reduces atherosclerosis in experimental models. This study investigated the association between p45 IgG autoantibodies and plaque inflammation in subjects with advanced cardiovascular disease., Methods: Native and MDA-p45 IgG levels were analyzed by ELISA in 349 carotid endarterectomy patients. In a subcohort of 195 subjects, endarterectomy samples were analyzed by immunohistochemistry and ELISA to determine plaque constituents and inflammation. Peripheral blood mononuclear cells were isolated from healthy donors., Results: Patients with preoperative events of neurological ischemia had lower levels of native p45 IgG. Low levels of MDA-p45 IgG were associated with increased risk of postoperative cardiovascular death during a mean follow-up of 54 months. High plasma levels of native p45 IgG were associated with increased plaque content of collagen and smooth muscle cell growth factors, as well as with lower levels of proinflammatory cytokines. Exposure of peripheral blood mononuclear cells from healthy donors to recombinant MDA-p45 IgG in presence of oxidized low-density lipoprotein reduced the expression of tumor necrosis factor-α and stimulated release of smooth muscle cell growth factors., Conclusions: This study confirms previous experimental findings of anti-inflammatory properties of apolipoprotein B-100 p45 antibodies and provides the first clinical evidence of associations between p45 IgG autoantibody levels and atherosclerotic plaque inflammation, plaque repair as well as prevalent and incident cardiovascular events in carotid endarterectomy patients. These findings suggest the possibility that treatment with anti-p45 antibodies may have beneficial effects in advanced cardiovascular disease., (© 2016 American Heart Association, Inc.)

Objective: Previous smaller studies have indicated inverse associations between autoantibodies to oxidized low-density lipoprotein epitopes, and cardiovascular disease. The present study investigated associations between autoantibodies against the apolipoprotein B-100 peptides p45 and p210, respectively, and risk of incident cardiovascular disease in a large population-based cohort., Approach and Results: Apolipoprotein B-100 autoantibodies were analyzed by ELISA in a prospective study, including 5393 individuals (aged 46-68 years) belonging to the cardiovascular arm of the Malmö Diet and Cancer study with a follow-up time of >15 years. Subjects that suffered an acute coronary event during follow-up (n=382) had lower levels at baseline of IgM autoantibodies recognizing the native and malondialdehyde-modified apolipoprotein B-100 peptides p45 and p210 and also lower IgG levels recognizing native p210, whereas no association was found with risk for stroke (n=317). Subjects in the highest compared with lowest tertile of IgM-p45MDA (hazard ratio [95% confidence interval]: 0.72 [0.55, 0.94]; P=0.017) and IgG-p210native (hazard ratio [95% confidence interval]: 0.73 [0.56, 0.97]; P=0.029) had lower risk for incident coronary events after adjustment for cardiovascular risk factors in Cox proportional hazard regression models. Moreover, subjects with high levels of IgG-p210native were less likely to have carotid plaques as assessed by ultrasonography at baseline (odds ratio=0.81, 95% confidence interval 0.70-0.95, P=0.008 after adjustment for risk factors)., Conclusions: This large prospective study demonstrates that subjects with high levels of apolipoprotein B-100 autoantibodies have a lower risk of coronary events supporting a protective role of these autoantibodies in cardiovascular disease., (© 2016 American Heart Association, Inc.)

Increased glucose levels are associated with the generation of advanced glycation endproduct (AGE) modifications. Interaction between AGE-modified plaque components and immune cells is believed to have an important role in the development of vascular complications in diabetes. Methylglyoxal (MGO) is one type of reactive aldehyde that gives rise to AGE modification. The present study analyzed whether autoantibodies against MGO-modified epitopes of the low-density lipoprotein apolipoprotein B (apoB) 100 predict cardiovascular events. A library consisting of 302 peptides comprising the complete apoB100 molecule was screened to identify peptides targeted by MGO-specific autoantibodies. Peptide (p) 220 (apoB amino acids 3286-3305) was identified as a major target. Baseline IgM and IgG against MGO-peptide 220 (p220) were measured in 700 individuals from the Malmö Diet and Cancer Cohort. A total of 139 cardiovascular events were registered during the 15-y follow-up period. Controlling for major cardiovascular risk factors demonstrated that subjects in the lowest tertile of MGO-p220 IgM had an increased risk for cardiovascular events (hazard ratio [95% confidence interval]: 2.07 [1.22-3.50]; p(trend) = 0.004). Interestingly, the association between MGO-p220 IgM and cardiovascular events remained and even tended to become stronger when subjects with prevalent diabetes were excluded from the analysis (2.51 [1.37-4.61]; p(trend) = 0.002). MGO-p220 IgM was inversely associated with blood glucose, but not with oxidized low-density lipoprotein. Finally, we demonstrate that anti-MGO-p220 IgM is produced by B1 cells. These data show that subjects with low levels of IgM recognizing MGO-modified p220 in apoB have an increased risk to develop cardiovascular events and that this association is present in nondiabetic subjects., (Copyright © 2015 by The American Association of Immunologists, Inc.)

Background: Model repositories such as BioModels Database provide computational models of biological systems for the scientific community. These models contain rich semantic annotations that link model entities to concepts in well-established bio-ontologies such as Gene Ontology. Consequently, thematically similar models are likely to share similar annotations. Based on this assumption, we argue that semantic annotations are a suitable tool to characterize sets of models. These characteristics improve model classification, allow to identify additional features for model retrieval tasks, and enable the comparison of sets of models., Results: In this paper we discuss four methods for annotation-based feature extraction from model sets. We tested all methods on sets of models in SBML format which were composed from BioModels Database. To characterize each of these sets, we analyzed and extracted concepts from three frequently used ontologies, namely Gene Ontology, ChEBI and SBO. We find that three out of the methods are suitable to determine characteristic features for arbitrary sets of models: The selected features vary depending on the underlying model set, and they are also specific to the chosen model set. We show that the identified features map on concepts that are higher up in the hierarchy of the ontologies than the concepts used for model annotations. Our analysis also reveals that the information content of concepts in ontologies and their usage for model annotation do not correlate., Conclusions: Annotation-based feature extraction enables the comparison of model sets, as opposed to existing methods for model-to-keyword comparison, or model-to-model comparison.

Objectives: Autoimmune responses against oxidized-LDL (oxLDL) have been suggested to modulate inflammation in atherosclerosis. Previous studies showed an association between autoantibodies against the apolipoprotein B (apoB) p210 antigen and a lower risk of cardiovascular (CV) events. In the present study we investigated if autoantibodies against p210 at the time of carotid endarterectomy (CEA) predict risk for future CV events., Methods: Native (nat) and malondialdehyde (MDA)-modified apoB p210 autoantibodies (IgM-p210nat, IgG-p210nat, IgM-p210MDA and IgG-p210MDA) were analyzed by ELISA from plasma samples of 351 patients at the time they underwent CEA. The incidence of postoperative CV events was assessed using national registers., Results: A total of 52 non-fatal and 15 fatal CV events were registered during the follow-up period (35.1 ± 16.7 months). Patients who suffered from a fatal CV event had significantly lower plasma levels of IgG-p210nat and IgG-p210MDA. Kaplan-Meier curves of event-free survival showed increased CV mortality in patients with levels of IgG-p210nat and IgG-p210MDA below the median (Log Rank 7.813, p .005 and 9.105, p .003 respectively). The association between low levels of p210 IgG and fatal post-operative CV events remained significant when adjusting for age, sex, total cholesterol, HDL cholesterol, smoking habits and hypertension in a Cox Proportional Hazard model (hazard ratios (HR) IgG-p210nat below median: HR 6.7 (95% C.I. 1.5-30.6, p .013) and IgG-p210MDA below median: HR 7.8 (95% C.I. 1.7-35.5, p .008)., Conclusions: The present findings support the notion that autoantibodies against LDL antigens are involved in the atherosclerotic disease process and suggest that CEA patients with low levels of IgG-p210nat and IgG-p210MDA have an increased risk of post-operative CV death., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Objectives: Immune responses against oxidized low density lipoprotein (LDL) play a key role in atherosclerosis. Previous studies have indicated inverse associations between autoantibodies to epitopes in oxidized LDL and cardiovascular disease. In this study we investigated the associations between autoantibodies against the apolipoprotein B-100 (apoB-100) peptides p45 and p210 and occurrence of carotid plaques., Design: The study cohort consisted of a population-based sample of 64-year-old women with varying degrees of glucose tolerance (n=594). To identify and record the occurrence of carotid atherosclerotic plaques ultrasonography was used. Measurements of plasma IgM and IgG autoantibodies against the native and malondialdehyde (MDA)-modified apoB-100 peptides p45 and p210 were performed by ELISA., Results: Women with carotid plaques were found to have lower levels of IgM MDA-p210 autoantibodies compared to plaque-free women. The number of carotid plaques in each subject and the total carotid plaque area correlated inversely with IgM MDA-p210 levels (r=-0.11, P=0.009 and r=-0.11, P=0.013, respectively). Furthermore, levels of IgM MDA-p210 above the lowest tertile were associated with an odds ratio of 0.55 (95% CI 0.38-0.79, P=0.001) for occurrence of carotid plaques, independently of other risk markers and statin treatment. Associations between apo-B100 peptide autoantibodies and cardiovascular risk factors were generally weak but subjects with impaired glucose tolerance had higher levels of IgM against MDA-p210., Conclusion: The present study demonstrates that high levels of IgM against MDA-p210 are associated with less severe carotid disease in women. These findings provide additional support for a role of immune responses against oxidized LDL in cardiovascular disease.

Objective: IL-25 has been implicated in the initiation of type 2 immunity and in the protection against autoimmune inflammatory diseases. Recent studies have identified the novel innate lymphoid type 2 cells (ILC2s) as an IL-25 target cell population. The purpose of this study was to evaluate if IL-25 has any influence on atherosclerosis development in mice., Methods and Results: Administration of 1 μg IL-25 per day for one week to atherosclerosis-prone apolipoprotein (apo)E deficient mice, had limited effect on the frequency of T cell populations, but resulted in a large expansion of ILC2s in the spleen. The expansion was accompanied by increased levels of anti-phosphorylcholine (PC) natural IgM antibodies in plasma and elevated levels of IL-5 in plasma and spleen. Transfer of ILC2s to apoE deficient mice elevated the natural antibody-producing B1a cell population in the spleen. Treatment of apoE/Rag-1 deficient mice with IL-25 was also associated with extensive expansion of splenic ILC2s and increased plasma IL-5, suggesting ILC2s to be the source of IL-5. Administration of IL-25 in IL-5 deficient mice resulted in an expanded ILC2 population, but did not stimulate generation of anti-PC IgM, indicating that IL-5 is not required for ILC2 expansion but for the downstream production of natural antibodies. Additionally, administration of 1 μg IL-25 per day for 4 weeks in apoE deficient mice reduced atherosclerosis in the aorta both during initiation and progression of the disease., Conclusions: The present findings demonstrate that IL-25 has a protective role in atherosclerosis mediated by innate responses, including ILC2 expansion, increased IL-5 secretion, B1a expansion and natural anti-PC IgM generation, rather than adaptive Th2 responses.

Avibactam is a novel non-β-lactam β-lactamase inhibitor that inhibits a wide range of β-lactamases. These include class A, class C, and some class D enzymes, which erode the activity of β-lactam drugs in multidrug-resistant pathogens like Pseudomonas aeruginosa and Enterobacteriaceae spp. Avibactam is currently in clinical development in combination with the β-lactam antibiotics ceftazidime, ceftaroline fosamil, and aztreonam. Avibactam has the potential to be the first β-lactamase inhibitor that might provide activity against class C-mediated resistance, which represents a growing concern in both hospital- and community-acquired infections. Avibactam has an unusual mechanism of action: it is a covalent inhibitor that acts via ring opening, but in contrast to other currently used β-lactamase inhibitors, this reaction is reversible. Here, we present a high-resolution structure of avibactam bound to a class C β-lactamase, AmpC, from P. aeruginosa that provided insight into the mechanism of both acylation and recyclization in this enzyme class and highlighted the differences observed between class A and class C inhibition. Furthermore, variants resistant to avibactam that identified the residues important for inhibition were isolated. Finally, the structural information was used to predict effective inhibition by sequence analysis and functional studies of class C β-lactamases from a large and diverse set of contemporary clinical isolates (P. aeruginosa and several Enterobacteriaceae spp.) obtained from recent infections to understand any preexisting variability in the binding pocket that might affect inhibition by avibactam., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

AZD5099 (compound 63) is an antibacterial agent that entered phase 1 clinical trials targeting infections caused by Gram-positive and fastidious Gram-negative bacteria. It was derived from previously reported pyrrolamide antibacterials and a fragment-based approach targeting the ATP binding site of bacterial type II topoisomerases. The program described herein varied a 3-piperidine substituent and incorporated 4-thiazole substituents that form a seven-membered ring intramolecular hydrogen bond with a 5-position carboxylic acid. Improved antibacterial activity and lower in vivo clearances were achieved. The lower clearances were attributed, in part, to reduced recognition by the multidrug resistant transporter Mrp2. Compound 63 showed notable efficacy in a mouse neutropenic Staphylococcus aureus infection model. Resistance frequency versus the drug was low, and reports of clinical resistance due to alteration of the target are few. Hence, 63 could offer a novel treatment for serious issues of resistance to currently used antibacterials.

Objective: Accumulating evidence shows that immune cells play an important role in atherosclerosis. Most attention has focused on the role of different T cell subsets, whereas the possible involvement of B cells has been less studied. In this study, we assessed the association of 2 different B cell subsets, CD19(+)CD40(+) and CD19(+)CD86(+) B cells, with risk for development of acute cardiovascular events., Approach and Results: The prospective study included 700 subjects randomly selected from the cardiovascular cohort of the Malmö Diet and Cancer study. Mononuclear leukocytes, stored at -140(○)C at the baseline investigation in 1991-1994, were thawed and B cell subsets analyzed by flow cytometry. Cytokine release from CD3/CD28-stimulated mononuclear leukocytes was measured with multiplex ELISA. Baseline carotid intima-media thickness and stenosis were assessed by ultrasonography, and clinical events were monitored through validated national registers during a median/mean follow-up time of 15 years. The subjects in the highest tertile of CD19(+)CD40(+) B cells had a significantly lower risk of incident stroke after adjustment for other risk factors. In contrast, CD19(+)CD86(+) B cells were associated with higher risk for development of a stroke event and increased release of proinflammatory cytokines from mononuclear leukocytes., Conclusions: These observations provide evidence for an involvement of B cells in the incidence of stroke and suggest that both pathogenic and protective B cell subsets exist.

Background: Esophageal dysmotility and gastroparesis are common secondary complications in patients with diabetes mellitus. Patients with dysmotility express antibodies against gonadotropin-releasing hormone (GnRH) in serum. The aim of the present study was to scrutinize patients with diabetes mellitus with regard to the presence of GnRH antibodies, and to examine associations between antibodies and clinical findings., Results: Thirty-nine consecutive patients with diabetes mellitus were included in the study after clinical examination and examination by esophageal manometry and gastric emptying scintigraphy. Serum was analyzed for the presence of antibodies against GnRH using an ELISA, and values are expressed as relative units (RU). Two age- and gender-matched healthy subjects per each patient served as controls. The prevalence of IgM GnRH antibodies in patients was 33% compared to 14% in controls (p = 0.027), with a higher antibody titer; 1.2 (0.6-5.0) and 0.2 (0.1-0.3) RU, respectively (p = 0.000). The expression of IgG antibodies was 15% in patients and none in controls (p = 0.000). Lower body mass index was associated with the presence of IgM antibodies (OR = 0.835, 95% CI = 0.699-0.998), and autonomic neuropathy with the presence IgG antibodies (OR = 9.000, 95% CI = 1.327-61.025). Esophageal dysmotility (69%) or gastroparesis (18%) were not associated with the presence of IgM antibodies (OR = 0.589, 95% CI = 0.143-2.424 and OR = 3.407, 95% CI = 0.633-18.350, respectively). Neither was esophageal dysmotility associated with IgG antibodies (OR = 2.500, 95% CI = 0.259-24.096)., Conclusions: Antibodies against GnRH are more common in patients with diabetes mellitus compared with healthy controls. IgM antibodies are associated with lower body mass index and IgG antibodies are associated with autonomic neuropathy.

Lipopolysaccharide (LPS) biosynthesis is an attractive antibacterial target as it is both conserved and essential for the survival of key pathogenic bacteria. Lipid A is the hydrophobic anchor for LPS and a key structural component of the outer membrane of Gram-negative bacteria. Lipid A biosynthesis is performed in part by a unique zinc dependent metalloamidase, LpxC (UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase), which catalyzes the first non-reversible step in lipid A biosynthesis. The UDP portion of the LpxC substrate-binding pocket has been relatively unexplored. We have designed and evaluated a series of hydroxamate based inhibitors which explore the SAR of substitutions directed into the UDP pocket with a range of substituted α-amino acid based linkers. We also provide the first wild type structure of Pseudomonas aeruginosa LpxC which was utilized in the design of many of these analogs., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Gonadotropin-releasing hormone (GnRH) analogs are given to women undergoing in vitro fertilization. Case reports describing the development of chronic intestinal pseudo-obstruction and auto-antibodies against GnRH after such treatment suggest a strong association between intestinal dysfunction and GnRH analogs. No experimental model for studying such a relationship is currently at hand. Our main goal was to investigate possible enteric neurodegeneration and titers of GnRH antibodies in response to repeated administration of the GnRH analog buserelin in rat. Rats were treated for 1-4 sessions with daily subcutaneous injections of buserelin or saline for 5 days, followed by 3 weeks of recovery. Buserelin treatment caused significant loss of submucous and myenteric neurons in the fundus, ileum, and colon. The loss of enteric neurons can, at least partly, be explained by increased apoptosis. No GnRH- or GnRH-receptor-immunoreactive (IR) enteric neurons but numerous luteinizing hormone (LH)-receptor-IR neurons were detected. After buserelin treatment, the relative number of enteric LH-receptor-IR neurons decreased, whereas that of nitric-oxide-synthase-IR neurons increased. No intestinal inflammation or increased levels of circulating interleukins/cytokines were noted in response to buserelin treatment. Serum GnRH antibody titers were undetectable or extremely low in all rats. Thus, repeated administrations of buserelin induce neurodegeneration in rat gastrointestinal tract, possibly by way of LH-receptor hyperactivation. The present findings suggest that enteric neurodegenerative effects of GnRH analog treatment in man can be mimicked in rat. However, in contrast to man, no production of GnRH auto-antibodies has been noted in rat.

Patients with functional gastrointestinal disorders express antibodies against gonadotropin-releasing hormone (GnRH) in serum. One common cause of posterior laryngitis (PL) is extra-esophageal reflux, but a functional etiology has also been suggested. The aim of this study was to scrutinize patients with PL with regard to the presence of GnRH antibodies and to examine the association between antibodies and symptoms and reflux. Consecutive PL patients were included after examination. Serum was analyzed for the presence of antibodies using an enzyme-linked immunosorbent assay (ELISA) method and expressed as relative units (RU). Two age- and gender-matched healthy subjects per case served as controls. The prevalence of IgM GnRH antibodies in patients was 35% compared with 28% in controls (P = 0.06), with higher levels in patients (0.8 (0.3-2.2) RU) than in controls (0.2 (0.1-0.6) RU) (P = 0.007). The corresponding IgG antibody prevalences were 43% and 4%, respectively (P = 0.001), with no difference in levels (P = 0.70). There was no association between antibodies and clinical findings.

Objective: Many patients, especially women, suffer from severe gastrointestinal pain and dysmotility for several years without being diagnosed. Depletion of gonadotropin-releasing hormone (GnRH) in the enteric nervous system (ENS) has been described in some patients. The aim of this study was to examine the expression of GnRH in ENS and antibodies against GnRH in serum, in a dysmotility patient cohort of southern Sweden., Materials and Methods: All consecutive patients (n = 35) referred for laparoscopic full-thickness biopsy because of symptoms or signs of severe dysmotility between 1998 and 2009, or patients with a severe dysmotility disorder having had a bowel resection within the time frame, were considered for inclusion. In 22 cases, representative biopsy material containing ganglia was available, and these patients were included. Medical records were scrutinized. The expression of GnRH was determined by immunohistochemistry in bowel biopsies from these patients and in patients with carcinoma or diverticulosis without ENS histopathology. Antibodies against GnRH in serum were determined by ELISA in patients and controls., Results: 14 patients were diagnosed with enteric dysmotility (ED) and 8 with chronic intestinal pseudo-obstruction due to varying etiology. Immunostained biopsies showed expression of GnRH in the ENS. A reduced expression of GnRH-containing neurons was found in 5 patients, as well as antibodies against GnRH in serum. 3 of these patients had a history of in vitro fertilization (IVF) using GnRH analogs., Conclusions: A subgroup of patients with severe dysmotility had a reduced expression of GnRH-containing neurons in the ENS and expressed antibodies against GnRH in serum.

Background: Diabetes is associated with increased cardiovascular disease, but the underlying cellular and molecular mechanisms are poorly understood. One proposed mechanism is that diabetes aggravates atherosclerosis by enhancing plaque inflammation. The Akita mouse has recently been adopted as a relevant model for microvascular complications of diabetes. Here we investigate the development of atherosclerosis and inflammation in vessels of Akita mice on LDLr⁻/⁻ background., Methods and Results: Akita-LDLr⁻/⁻ and LDLr⁻/⁻ mice were fed high-fat diet from 6 to 24 weeks of age. Blood glucose levels were higher in both male and female Akita-LDLr⁻/⁻ mice (137% and 70%, resp.). Male Akita-LDLr⁻/⁻ mice had markedly increased plasma cholesterol and triglyceride levels, a three-fold increase in atherosclerosis, and enhanced accumulation of macrophages and T-cells in plaques. In contrast, female Akita-LDLr⁻/⁻ mice demonstrated a modest 29% increase in plasma cholesterol and no significant increase in triglycerides, atherosclerosis, or inflammatory cells in lesions. Male Akita-LDLr⁻/⁻ mice had increased levels of plasma IL-1β compared to nondiabetic mice, whereas no such difference was seen between female diabetic and nondiabetic mice., Conclusion: Akita-LDLr⁻/⁻ mice display considerable gender differences in the development of diabetic atherosclerosis. In addition, the increased atherosclerosis in male Akita-LDLr⁻/⁻ mice is associated with an increase in inflammatory cells in lesions.